A terrible disease of the cardiovascular system, atherosclerosis, develops in the areas of bends andbranches of arteries, where the direction and modulus of the blood flow velocity vector change, and consequentlyso does the mechanical effect on endothelial cells in contact with the blood flow. The review focuses on topicalresearch studies on the development of atherosclerosis – mechanobiochemical events that transform the proatherogenicmechanical stimulus of blood flow – low and low/oscillatory arterial wall shear stress in the chains of biochemicalreactions in endothelial cells, leading to the expression of specific proteins that cause the progressionof the pathological process. The stages of atherogenesis, systemic risk factors for atherogenesis and its importanthemodynamic factor, low and low/oscillatory wall shear stress exerted by blood flow on the endothelial cells liningthe arterial walls, have been described. The interactions of cell adhesion molecules responsible for the developmentof atherosclerosis under low and low/oscillating shear stress conditions have been demonstrated. The activationof the regulator of the expression of cell adhesion molecules, the transcription factor NF-κB, and the factorsregulating its activation under these conditions have been described. Mechanosensitive signaling pathways leadingto the expression of NF-κB in endothelial cells have been described. Studies of the mechanobiochemical signalingpathways and interactions involved in the progression of atherosclerosis provide valuable information for thedevelopment of approaches that delay or block the development of this disease.Key words: atherogenesis; shear stress; transcription factor NF-κB; RelA expression; mechanosensitive receptors;cell adhesion molecules; signaling pathways; mechanotransduction. Nowadays, cardiovascular disease is a major public healthissue. Moreover, atherosclerosis is one of the most commonpathologies of the cardiovascular system. The systemic riskfactors for the development of atherosclerosis include age,hypertension, diabetes mellitus, smoking, low physical activity,fatty diet, renal failure, increased level of fibrinogen, lowdensitylipoproteins, cholesterol and blood plasma C-reactiveprotein . The level of low-density lipoproteins(LDLs) is classified into a separate group of factorsthat account for the atherogenicity of the subfraction profileof apo-B-containing lipoproteins . The penetration of blood plasma LDLs throughthe endothelium in athero-susceptible areas of the arteries andtheir retention and accumulation in the extracellular matrix(ECM) of the subendothelial space initiates atherogenesis.LDLs are retained in the intima (mainly due to interaction withproteoglycans), undergo oxidation (formation of oxLDLs) andcause an inflammatory response – the infiltration of circulatingblood monocytes into the intima. In the intima, monocytesdifferentiate into macrophages, uptake oxLDLs and becomefoam cells .The development of atherosclerosis occurs in the followingstages: (i) adaptive intimal thickening, (ii) formation of fattystreaks, (iii) pathological intimal thickening (PIT), (iv) earlyfibroatheroma and (v) late fibroatheroma. In stage (i), smoothmuscle cells (SMCs) of the media migrate to the intima and secreteproteoglycans. The formation of fatty streaks in stage (ii)is accompanied by the accumulation of foamy cells (macrophagesloaded with lipids) in the intima. Lipid-loaded SMCsare less represented. The PIT process (iii) occurs with andwithout the infiltration of macrophages. In both cases, SMCsand extracellular lipid pools are present in the intima. The accumulationof SMCs occurs towards the lumen of the artery,and lipid pools accumulate close to the media. The formationof a fibrous cap that covers the necrotic core occurs in the laterstages of development of the atherosclerotic lesions, includingearly and late fibroatheroma . The cap includes SMCs,infiltrated macrophages, T-lymphocytes, as well as collagensand proteoglycans of the extracellular matrix. Programmedcell death, via apoptosis and necroptosis, plays an essentialrole in early fibroatheroma (iv) with the formation of foci ofnecrosis and cholesterol crystals. In late fibroatheroma (v),an extensive necrotic core that consists of cellular debris anda large number of crystals of free cholesterol and its esters isformed .The molecular and genetic processes of atherogenesis remainunclear. Numerous haemodynamic studies have shownthat, based on systemic risk factors, atherosclerosis developsmainly in the bends and branching of the arteries, where thereis a change in the nature of the blood flow . The haemodynamiccharacteristics of the effect of blood flow on the vessel wallsare wall shear stress (WSS), hydrostatic pressure and cyclicdeformation. WSS is the friction force that occurs whenflowing blood comes into contact with the inner wall of the artery. WSS on the arterial wall is described by :where μ is the blood viscosity index, →V(t) is the blood flowrate parallel to the vessel wall at time t and r is the radial coor-dinate.Studies conducted on animal models and observing patientsrevealed a regular maximum thickening of the intima and theformation of atherosclerotic plaques in areas with low WSS(< 10 dyn/cm2 in humans) and low/oscillatory WSS (with adeviation of the instantaneous WSS vector from its averagedirection). Such damage was minimal in areas of high WSS(>25 dyn/cm2 in humans). High values of WSS were realizedin the rectilinear sections of the arteries with laminar bloodflow. In the areas of branching and bending of the arteries nearthe walls, vortex flows were formed, leading to mechanicalstress on the walls, which was accompanied by pathologicaleffects. These flows were characterised by low and low/oscillatoryWSS .A study conducted on isolated segments of blood vesselsthrough which LDLs flowed demonstrated that the transport ofLDLs into the vascular wall increased with a decrease in WSSand, on the contrary, decreased with an increase in WSS . Patient-specific modelling of the subendothelialaccumulation of LDLs in the stenotic right coronary arteryalso showed an inverse relationship between the distribution ofWSS and the accumulation of LDLs .The zone of recirculating flow and low WSS correspondedto the maximum accumulation of LDLs, and in areas of highWSS, the accumulation of LDLs was low.To localise the segments of arteries with low and low/oscillatoryWSS and monitor the transformation of atheroscleroticplaques into a stable or unstable phenotype, computer modellingof blood flow in the vessels is being developed. Thiswill facilitate the identification of patient-specific fields andgradients of blood flow rates depending on the geometry ofthe vessels . To solve these issues, the Navier–Stokesequations for an incompressible viscous fluid are used. Toreconstruct the geometric shape of the vessels, intravascularultrasound methods are used as well as X-ray microcomputertomography with the use of contrast agents . The ANSYS Fluent, OpenFOAM,FLUENT 6.0 and other software packages are widely used toconduct calculations via mathematical models of stationaryand unsteady blood flows in various areas of the arteries. Computermodelling of the distribution of WSS, which accountsfor patient-specific data on the geometry of blood vessels, isof high value for clinical practice.The molecules of cell adhesionand their interactions in the early stageof atherogenesisOxLDLs in the subendothelial space as well as low and low/oscillatory WSS cause pro-inflammatory activation of endothelialcells (ECs), which leads to the rolling of leukocytes inthe circulating blood flow to the endothelium, their adhesion and transendothelial migration (TEM). The mediators of thesecritical events of the early stage of atherogenesis are cell adhesionmolecules. These molecules are expressed on the surfaceof ECs and circulating blood cells (monocytes or leukocytesand platelets) and include platelet endothelial cell adhesionmolecule-1 (PECAM-1); intercellular adhesion molecule-1and -2 ; vascular cell adhesion molecule-1(VCAM-1); E-, L- and P-selectins; vascular endothelial (VE)cadherin; β1 and β2 integrins; proline-rich glycoprotein CD99and junctional adhesion molecule-A (JAM-A).Selectins (transmembrane glycoproteins) are expressed onthe surface of ECs , leukocytes(L-selectin) and platelets (P-selectin) . Early experiments conducted in a flow chamber with alaminar flow of monocytes on an EC monolayer  showed that the rolling of the monocytes to theECs, weak, reversible contact of the monocytes with ECs and slowing down of the rate along theendothelium determine the interactions of the L-selectin ofthe monocytes with glycoprotein ligands of the ECs and, to alesser extent, the P-selectin of the ECs with the glycoproteinligand PSGL-1 of the monocytes. E-selectin is not involvedin the process . Firm, irreversibleadhesion of the leukocytes to the ECs occurs during theinteraction of the leukocyte α4β1 (VLA-4) integrin with theendothelial immunoglobulin VCAM-1  and the interaction of the leukocyteαLβ2  integrinswith the endothelial immunoglobulin ICAM-1 .OxLDLs and lysophosphatidylcholine (a component ofoxLDLs) induce the expression of ICAM-1 and VCAM-1 onthe surface of cultured ECs and stimulate monocyte adhesion. A physiologicallylow WSS, created by the flow of leukocytes  on the EC monolayer,generates upward docking structures on the endothelium thatcontain ICAM-1 and VCAM-1 clusters within 1–2 minutes.These clusters surround the leukocytes and function as ananchor for them .In turn, the structures of VCAM-1 and ICAM-1 that surroundthe leukocytes stimulate the formation of lateral linear tracksof leukocyte β1 (VLA-4) and β2 (LFA-1) integrins that areoriented parallel to the ICAM-1 and VCAM-1 clusters . Moreover, 90% of the leukocytes thatare surrounded by the VCAM-1 and ICAM-1 clusters transmigrateto the subendothelial space, and the suppression ofVCAM-1 and ICAM-1 via inhibitors  significantly suppresses TEM. Regardless of theTEM pathway , the TEM process isassociated with the formation of a cupped traction structureby the VCAM-1/VLA-4 and ICAM-1/LFA-1 interactions ofendothelial and leukocyte cells, which guide and facilitateTEM .VE-cadherin plays an important role in the TEM of leukocytes.VE-cadherin is only expressed in ECs, localised mainlyin the intercellular contacts and plays an important role in the intercellular adhesion and barrier functions of the ECs . The adhesion of leukocytes to the endothelium inthe lateral intercellular contacts, preceding TEM, induces theformation of gaps in the intercellular distribution of VE-cadherinand the components of the VE-cadherin complex  at the sites of adhesion or transmigrationof the leukocytes. The gaps are formed as a result of lateraldisplacement of VE-cadherin in the membrane and facilitatethe TEM of the leukocytes. Following the completion of TEM,VE-cadherin moves in the opposite direction and closes thegaps (curtain opening and closing effect) . The lateral displacement of VE-cadherinin the membrane most likely occurs due to the destabilisationof the VE-cadherin bond with the actin cytoskeleton by thefollowing mechanism: the ICAM-1 and VCAM-1 clusters inthe sites of intercellular adhesion of the leukocytes to the ECsinduce the intracellular activation of the Src and Pyk2 tyrosinekinases and the phosphorylation of Tyr658 and Tyr731 of thecytoplasmic domain of VE-cadherin, which are involved in thelow-affinity binding of VE-cadherin to p120- and β-catenin,respectively. The weakening of these interactions disrupts theVE-cadherin bond with the actin cytoskeleton, destabilises theVE-cadherin or VE-cadherin cell-cell interactions and facilitatesthe lateral movement of the phosphorylated VE-cadherinin the membrane . p120-catenin regulatesthe phosphorylation of VE-cadherin and the paracellularTEM of leukocytes via a competition mechanism with theactivated Src and Pyk2 tyrosine kinases: the overexpressionof p120-catenin in the ECs leads to the absence of gaps in thedistribution of VE-cadherin and the blocking of the TEM ofthe leukocytes .The glycoprotein PECAM-1 (CD31) plays an importantrole in the TEM of leukocytes. In ECs, PECAM-1 is mainlylocalised in the intercellular contacts. The homophilic interactionsof this protein with adjacent ECs occur through theextracellular Ig-like domains IgD1 and IgD2 . PECAM-1 is also present in a distinct membranevesicularrecycling compartment adjacent to the lateral bordermembrane of the ECs . In the restingendothelium (in the absence of adhesive leukocytes), there isconstitutive membrane traffic between the lateral cell borderand the membrane-vesicular compartment  . In the presence of adhesive leukocytes onthe endothelium, directed kinesin-dependent migration of thePECAM-1-bearing LBRC membrane along the microtubulesof the ECs to the sites of para- and transmigration of the leukocytesoccurs, in addition to the surrounding of the leukocytesby the LBRC membrane. The LBRC provides non-ligatedPECAM-1 and CD99 in the ECs to interact with the homophilicligands  in the leukocytesand initiates the signals for further recruitment of theLBRC as the leukocytes move through the endothelial layer.The antibodies to PECAM-1 and CD99 block the TEM of theleukocytes . The recruitment ofthe LBRC to paracellular TEM sites precedes the formationof gaps in the intercellular distribution of VE-cadherin andis necessary for the formation of these gaps . In ApoE–/– PECAM-1–/– mice, the load of plaques inthe areas of the carotid artery with a low and low/oscillatory WSS was significantly less than in the control ApoE–/– mice. A study on the relationship of singlenucleotide polymorphisms  in functionally important domainsof PECAM-1 in patients who were at risk of developingcoronary heart disease and myocardial infarction showed thatArg670Gly substitution can be a homozygous protector forthe development of myocardial infarction. This substitution islocalised close to Tyr663, the phosphorylation of which, underlow WSS conditions, initiates the signaling pathway of activationof the key transcription factor NF-κB for the expressionof cell adhesion molecules . Val125Leuand Asn563Ser substitutions are not associated with the riskof coronary heart disease .Integrins also play an important role in cell adhesion. Theintegrins are a large family of receptors that are localised inthe plasma membrane and consist of 18 α and 8 β subunitsthat form 24 different heterodimers. The extracellular domainsof the integrins interact with ECM proteins  and ligands on the surface of other typesof cells, causing cell-substratum or cell-cell adhesion. Theintegrin-ligand interactions induce the activation of a variety ofsignaling pathways that modulate cellular behaviour, includingproliferation, shape, motility, survival or apoptosis, differentiation,protein phosphorylation, cytoskeleton organisation andgene expression. Many integrins are expressed in an inactivestate on the cell surface since the membrane-proximal highlyconserved sequences of the cytoplasmic domains of the αand β subunits form a structural constraint that locks the conformationof integrins in an inactive, low-affinity state. Theactivation of integrins is often induced by intracellular signalsand regulatory factors that act on the cytoplasmic domains, aswell as phosphorylation. This alters the affinity of integrins forligands through conformational changes in their extracellulardomains, as well as clustering .The adhesion of leukocytes to the endothelium and theirinfiltration into the subendothelial space is enhanced bycytokines, chemokines and other factors. Thus, monocyticchemotactic protein-1 (MCP-1)  and interleukin-8 (IL-8) (producedby macrophages) are involved in the delay of peripheralcirculation monocytes and their adhesion and migration intothe arterial intima via interaction with monocyte receptors belongingto the CCR2 and CXCR2 types, respectively . IL-9, secreted mainly byCD45+CD3–CD19– leukocytes in АпоЕ–/– mice, stimulates theexpression of VCAM-1 in the ECs of the aorta of mice throughinteraction with the IL-9 receptor (IL-9R) and activation(phosphorylation) of the signaling protein and transcriptionactivator STAT3 . In the atheroscleroticaortas of АпоЕ–/– mice, a high level of IL-17A expressionwas observed, as well as numerous IL-17A-producing CD4+T helper 17 (Th17) and γδ+ T cells. IL-17A initiates the productionof several cytokines and chemokines by aortic cells,in particular, the pro-inflammatory chemokine CXCL1, whichactivates peripheral circulation monocytes and stimulates theiradhesion and migration to the aortic wall .Pro-atherogenic IL-17C, expressed mainly by aortic SMCsin АпоЕ–/– mice, is involved in the recruitment of T cells and macrophages into the aortic wall .TGF-β-(H2O2-) inducible clone 5 (His-5), expressed on thesurface of ECs and SMCs, is involved in the formation ofstructures that are similar to the microvilli on the surface ofECs, which enhance the adhesion of monocytes to the ECs.Transcription factor NF-κB is a key regulatorof the gene expression of cell adhesion moleculesunder conditions of physiologicallylow and low/oscillatory wall shear stressThe transcription factor NF-κB positively regulates the expressionof cell adhesion molecules with the participation ofother transcription factors and coactivators. The VCAM-1 genepromoter has two NF-κB sites that are required for transcriptionactivation . The C/EBP and NF-κBsites were identified in the ICAM-1 gene promoter, and mutationsin the latter completely suppress the activation of theICAM-1 promoter . The ELAM-1 genepromoter includes a CRE/ATF site, three NF-κB sites and threeHMGI(Y) sites; two of the HMG I(Y) sites are located withinthe NF-κB sites (the interaction of HMG I(Y) and NF-κBwith small and large DNA grooves, respectively). All threeNF-κB sites are required for promoter activation and enhancethe affinity of NF-κB and ATF-2 for the promoter . The promoter of the MCP-1 gene (monocytechemoattractant protein-1) contains the NF-κB site and theAP-1 site, which are necessary for maximum induction of thepromoter . The core element GAGACC(SSRE) was identified in the pro-atherogenic platelet growthfactor (PDGF) promoter that stimulates the proliferation andmigration of SMCs ; it was found tointeract with NF-κB .In ECs, the most common p50/p65 (RelA) heterodimer isNF-κB. It is well known that, in the cytoplasm, latent NF-κBis associated with an inhibitor of IκB (mainly IκBα) and isinactive. When ECs are stimulated by cytokines TNFα, IL-1 orbacterial lipopolysaccharide (LPS) that interact with specificEC receptors, signaling pathways are activated, leading to theactivation of the IκB kinase (IKK complex), which specificallyphosphorylates IκB. After the ubiquitination of IκB and itsproteasomal degradation, free NF-κB is translocated into cellnuclei (factor activation), interacts with elements of the DNAmajor groove and activates the expression of genes involved inthe immune response, cell survival, carcinogenesis and inflammation(with the participation of other transcription factorsand coactivators) .Early studies demonstrated that active NF-κB and ICAM-1were present in ECs, macrophages and SMCs in atheroscleroticplaques of arteries of deceased patients but were absentin the intima or media of the healthy arteries of those patients. OxLDL triggers in the ECs in vitro andin vivo signaling pathways for IKK complex activation; thisincludes the activation of focal adhesion kinase (FAK) andribosomal S6 kinase (RSK) and leads to NF-κB activation,VCAM-1 expression and monocyte adhesion to ECs . Using the model flow channel system  that is created by a laminar pulsating flow of theculture medium containing monocytes, in which an averagephysiologically low and uniformly distributed WSS acts on the EC monolayer, it was shown that a mechanical stimulus activates the IKK complex andNF-κB and stimulates VCAM-1 expression and monocyteadhesion to the ECs . A model spatialgradient of averaged low/oscillatory WSS, close to the gradientof WSS in arterial branchings and bends, resulted in amore efficient activation of NF-κB compared to uniformlydistributed low WSS .High activation of NF-κB was observed in ECs that werecultivated based on a model of the calculated WSS profile in asite of the human carotid sinus  compared with a model of the calculatedWSS profile in the distal segment of the carotid arterybifurcation  orcells at rest . A study of the transcriptionalexpression of the VCAM-1 endothelium from the nature ofhaemodynamics and calculated profiles of WSS in the segmentsof the left and right coronary arteries showed that low/oscillatory WSS at the outer wall of the bifurcation of theleft anterior descending artery (atherogenic region) inducesa significantly higher expression of VCAM-1 compared to astraight region of the right artery , inwhich atherosclerotic plaques are largely unformed . The mapping of the endothelial expression ofNF-κB/IκB and the activation of NF-κB in areas of the aorticarch of mice with high  and low  probability of atherosclerotic plaque formationdemonstrated that in the HP region with low/oscillatory WSS, the expression of NF-κB/IκB and the activationof NF-κB significantly exceeded those in the LP region. S. Cuhlmann et al. (2011) also detectedincreased expression of the RelA subunit NF-κB and a highernuclear localisation compared with the LP region in the HPregion of the aortic arch of mice. Moreover, this was correlatedwith increased expression of VCAM-1 and the accumulationof CD68+ macrophages in the HP regionThe gap-junction protein connexin 40 (Cx40), expressedin ECs under low WSS conditions, whose cytoplasmic C-terminusinteracts with IκBα and inhibits its phosphorylation,is a negative regulator of NF-κB activation in areas with lowWSS . Conversely, mechanosensitive phosphatidicacid phosphatase (PPAP2B) regulates NF-κB activity,expression of adhesion proteins, monocyte adhesion and TEMunder high WSS. PPAP2B shows increased expression inECs under atheroprotective flow characteristics  and shows decreased expression under atherogeniccharacteristics . PPAP2Bhydrolyses lysophosphatidic acid (LPA), a pro-atherogenicand thrombogenic glycerophospholipid in the blood and,thereby, blocks signaling pathways activated by the interactionof LPA with cellular LPA receptors (LPAR). LPA-LPAR1interaction activates the Rho kinase-NF-κB signaling pathwayand the subsequent transcriptional expression of ICAM-1,VCAM-1 and E-selectin in ECs .LPA-LPAR1/2 interaction leads to an increase in the contractilityof ECs and the permeability of the endothelial monolayer. LPA is an extracellular signalingmolecule that is capable of interacting with at least six G protein LPA-LPAR1/2 interaction leads to an increase in the contractilityof ECs and the permeability of the endothelial monolayer. LPA is an extracellular signalingmolecule that is capable of interacting with at least six G protein-coupled cellular receptors, initiating intracellular signalingcascades  and exerting multiple effectson blood cells  and vascular wall cells. Therefore, in platelets, LPA induces a changein the shape, aggregation, and formation of platelet-monocyteaggregates. In ECs, LPA induces cell migration, expressionof VCAM-1, ICAM-1, E-selectin, chemokines (CXCL1),formation of actin stress fibres and cell contraction. In SMCs,LPA induces cell contraction, migration and proliferation.Additionally, LPA accumulates in the lipid-rich core of atheroscleroticplaques, and, when ruptured, enters the bloodstreamand activates platelets, leading to the formation of blood clots. Endothelial NO synthase (eNOS) isalso expressed under physiologically high WSS. Under theseconditions, the eNOS promoter is activated through the interactionof NF-κB with the GAGACC element . eNOS produces NO, an inducer of IκBα expression.NO stabilises the NF-κB∙IκBα heterotrimer in the cytoplasmand induces IκBα translocation into cell nuclei, leading to theinactivation of NF-κB and the termination of NF-κB-mediatedtranscription . Moreover, NO inhibitsIκB kinase (IKK complex), a positive regulator of NF-κBactivation .Mechanosensitive signaling pathwaysthat control the transcriptional expressionof the RelA subunit of NF-κB in endothelial cellsThe WSS, created by the blood flow to the walls of the arteries,activates mechanosensitive signaling pathways in the ECs.Amechanosensor localised in the cell membrane perceives amechanical stimulus (WSS) and triggers intracellular signalingthat activates specific transcription factors, which regulatethe transcriptional expression of proteins. The internal bendof the aortic arch and the areas of arterial bifurcation, wherelow and low/oscillatory WSS are realized, are associated withthe localisation of atheroma. In the same areas, increased expressionof active c-Jun N-terminal kinase 1 (JNK1), whichbelongs to mitogen-activated protein kinases (MAPK), wasrevealed , as well as increased expres-sionand nuclear localisation of the RelA subunit of NF-κB. The hypothesis that physiologicallylow WSS regulates the expression of RelA and NF-κB targetgenes (VCAM-1 and others) through the JNK1-dependentpathway was proved by modelling the haemodynamics ofthe carotid artery in mice. The implantation of a tapered cuffaround the mouse carotid artery generates a laminar bloodflow with a low velocity and low WSS upstream of the cuffand disturbed, low velocity and low/oscillatory WSS downstreamof the cuff .This approach demonstrated that increased expression ofRelA and activated JNK1 is realised in regions with lowand low/oscillatory WSS. The transcription factor ATF2 isactivated by JNK1 (phosphorylation) and interacts with theRelA promoter sites to activate the promoter. Thus, in areaswith low/oscillatory WSS, the JNK-ATF2-RelA signalingpathway is implemented, which stimulates the expression ofRelA and NF-κB target genes (VCAM-1). The pathway alsostimulates the accumulation of CD68+ macrophages, whichare an indicator of the development of arterial inflammation.The signaling pathway for the transcriptional expressionof RelA upstream of JNK1 includes the activation ofintegrins, which is initiated by the stimulation of the cellsurface mechanosensory complex and an integrin-dependentsignaling cascade, leading to the activation of JNK1. The cellsurface mechanosensory complex includes the PECAM-1 andVE-cadherin receptors and the vascular endothelial growthfactor 2 receptor (VEGFR2), which belongs to the receptortyrosine kinase subfamily . In this complex,PECAM-1 is a key mechanosensitive signaling moleculethat perceives a mechanical signal and converts it into a chainof intracellular biochemical reactions. WSS, acting on theextracellular domain of PECAM-1, affects the conformationof the cytoplasmic domain of PECAM-1 and the availabilityof Tyr663 and Tyr686 of this domain for phosphorylation.Phosphorylation is conducted by membrane-bound Fyn tyrosinekinase (family of Src tyrosine kinases), which is localisedin intercellular contacts near PECAM-1 . PECAM-1/PECAM-1 intercellular EC interactionsthrough the extracellular domains of PECAM-1  are required for efficient phosphorylation and thetriggering of intracellular mechanosensitive signaling . Activated PECAM-1 andVE-cadherin (which functions as an adapter and is associatedwith VEGFR2) facilitate the phosphorylation of VEGFR2 atTyr801 and Tyr1175 by the Src tyrosine kinase. In turn, thephosphorylated VEGFR2, through direct interaction withthe regulatory p85 subunit of the phosphatidylinositol-3-OHkinase (PI(3)K), phosphorylates PI(3)K .The activated PI(3)K stimulates the conformational activationof integrins through the conservative pathway of theassociation of phosphatidylinositol-3,4,5 triphosphate (PI(3)Kproduct) with pleckstrin homology (PH) domains of cytohesin-1 or cytohesin-like proteins. This is followed by the translocationof these proteins to the plasma membrane and theirassociation with the cytoplasmic domains of the β subunitsof integrins, which is realized in various types of cells. Thisinteraction leads to conformational changes in the extracellulardomains of the α and β subunits of integrins and an increase intheir affinity for specific ECM proteins, as well as clustering. In athero-resistant areasof arteries, the ECM is rich in CL (IV) and LN. However, inatherogenic regions (low/oscillatory WSS conditions), theECM is enriched in the pro-inflammatory proteins FN andFG .Antibodies specific to ligated β1 and β3 integrins were usedto show that, under conditions of low/oscillatory WSS, thereis an increase in the binding of α5β1 integrin to FN  and of αvβ3 integrin to VN.Thus, the conformational activation of integrins and thedynamic formation of new bonds of integrins with specificligands occur – ECM proteins are realized .The composition of the ECM and the ligation of integrinsby the ECM proteins activate many intracellular signalingcascades – in this case, the Shc-Grb∙Sos-Ras-MAPKsignaling pathway. The cellular adapter protein Shc, activatedunder conditions of low/oscillatory WSS by tyrosinekinases Src and VEGFR2 close to the EC contacts (phosphorylationof Tyr239/240), forms an early unstable complex Shc∙VEGFR2∙VE-cadherin . Next, a stablecomplex Shc with an αvβ3 integrin that is ligated with FN orVN is formed . Additionally, Shc associates with the β1 and β5 integrinsthat are ligated with FN and VN . In this manner, Shc coordinates the intercellularcontact proteins (VE-cadherin) and integrin-ECM interactionsunder low/oscillatory WSS conditions. Membrane-associatedsmall G proteins Ras  function cyclicallybetween active Ras∙GTP and inactive Ras∙GDP forms, whichare a molecular switch of an intracellular signal in response toan extracellular stimulus . The complexesof phosphorylated Shc with the αvβ3, β1 and β5 integrinsformed on the cytoplasmic side of the plasma membrane areaccompanied by the association of Shc with the cytoplasmicGrb2∙Sos complex of the growth factor receptor-bound protein2 (Grb2) and the guanine nucleotide exchange factor Sos.This complex stimulates the rate of exchange of the GDPassociated with Ras, on the GTP  . In turn, activated Rasstimulates cytoplasmic Raf kinase  and recruits Raf to the inner surfaceof the plasma membrane through direct interaction with itsregulatory domains and the subsequent phosphorylation offour sites of the kinase domain . Raf,activated via a MAP kinase cascade, activates MAPK (JNK).However, the activation of Shc and membrane-associatedheterotrimeric G proteins leads to the stimulation of Ras under the conditionsof the action of WSS on the cells. In the absence of WSS,G proteins are activated via association with ligand-activatedreceptors: the α subunit exchanges the GDP bound to it forGTP and dissociates from the βϒ dimer. The α∙GTP and βϒcomplexes become mediators of cellular event signaling untilthe α subunit restores its inactive GDP-bound state (since ithas GTPase activity.) The reassociation of α∙GDP with the βϒdimer provides an inactive Gαβϒ heterotrimer that is capableof entering a new activation cycle . Alterationsin the physical properties of membranes under WSSconditions affect the conformation and functions of membrane-associatedproteins and, as a result, the signaling pathways activated bythese proteins .Purified heterotrimeric G proteins in phospholipid liposomesloaded with [ϒ-32P]GTP were shown to be activatedby the action of physiological levels of WSS on liposomes. Within 1 second, a physiologically lowWSS activated the Gαq/11 and Gαi3/0 subunits of G proteins inECs . The activated Gαq and Gβϒ subunitsdissociated from them under conditions of a WSS gradientin a physiologically low range and initiated Ras activationand downstream MAPK signaling .Moreover, two primary basic mechanosensors, Gαq/11 andPECAM-1, establish a mechanosensitive Gαq/11∙PECAM-1complex, which is formed under conditions of laminar flow in vivo (atheroprotective straight region of the descending aorta of mice) and in vitro. Underoscillatory flow conditions (low/oscillatory WSS), leading tothe activation of G proteins, this complex is rapidly (within30 seconds) destroyed . The formation ofthe Gαq/11∙PECAM-1 complex involves PECAM-1 extracellularIg-like domains 2 and 3, as well as a Gαq/11 interactingreceptor that associates with Gαq/11 and PECAM-1 Ig-likedomains 2 and 3 and serves as a bridge in the formation of thecomplex . The PECAM-1∙Gαq/11 complexalso includes heparan sulfate proteoglycan, which associateswith the Ig-like domain 3 of PECAM-1 and mediates theformation of the PECAM-1∙Gαq/11 complex .Computer modelling of the blood flow in the arteries makesit possible to determine the most atherogenic areas of thearteries, which are characterised by low and low/oscillatoryWSS. The transcription factor NF-κB and the cell adhesionmolecules ICAM-1, VCAM-1 and E-selectin are the earliestmarkers of atherogenesis. Therefore, the processes involved inthe expression of these proteins under the conditions inducedby a mechanical stimulus (low and low/oscillatory WSS) onendothelial cells are of great interest. This review presents ananalysis of numerous studies that demonstrated how the activationof membrane-bound proteins that perceive a mechanicalstimulus (low and low/oscillatory WSS) triggers a cascadeof biochemical reactions that lead to the transcriptional expressionof NF-κB, a key regulator of the expression of celladhesion molecules. This review also describes in detail themechanisms of interaction between the endothelial cell adhesionmolecules and blood leukocytes that are responsible foradhesion and the subsequent TEM of leukocytes during the initialstage of atherogenesis. 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Senegalia pennata subsp. insuavis (Lace) Maslin, Seigler & Ebinger, Citrus hystrix DC. and Solanum melongena ‘Kermit’ extracts exhibited high antioxidant activities. Moreover, Citrus hystrix DC. extract was a potent inhibitor against lipase, angiotensin-converting enzyme and butyrylcholinesterase, while Coriandrum sativum L. and Psophocarpus tetragonolobus (L.) DC. were potent anti-diabetic agents and Senegalia pennata subsp. insuavis (Lace) Maslin, and Seigler & Ebinger was a potent anti-glycation agent. Our data provide a comparative analysis of ten vegetables to encourage healthy food consumption and development to control NCDs in Thailand in the future.Non-communicable diseases (NCDs) are the leading global cause of death. The World Health Organization (WHO) has endorsed the consumption of fruits and vegetables because they are rich in phytochemicals that sustainably ameliorate the occurrence of NCDs. Thai food contains many spices and vegetables with recognized health benefits. Quality control of plant samples encountered a bottleneck in the field and comparative studies of plant control origins including species or cultivar identification, growing area and appropriate harvesting time are limited. To address this issue, all plant samples used in this study were cultivated and controlled by the Department of Agriculture, Ministry of Agriculture and Cooperatives, Thailand. The samples were phytochemically screened and determined their health-promoting bioactivities via antioxidant activities and inhibition of NCD-related enzymes including lipase (obesity), α-amylase and α-glucosidase (diabetes), angiotensin-converting enzyme (hypertension), as well as acetylcholinesterase, butyrylcholinesterase and β-secretase (Alzheimer’s disease). The non-enzymatic reaction toward glycation was also evaluated. The results showed that  Non-communicable diseases (NCDs) including cancer, diabetes mellitus (type II), cardiovascular diseases, hypertension, and Alzheimer’s disease (AD) are the leading cause of death worldwide. In 2012, the World Health Organization (WHO) reported that NCDs accounted for 68% of morbidity (38 million deaths from 56 million deaths), while over 16 million 40%) were early deaths (<70 years) [0% were eFruits and vegetables are rich in fiber, vitamins, minerals, and phytochemicals including alkaloids, anthocyanins, glucosinolates, flavonoids, phytosterols, phenolic acids and terpenoids that are secondary plant metabolites, with pharmacological effects toward a wide range of ailments including NCDs . PhytochAllium cepa Aggregatum Group, Allium fistulosum L., Allium sativum L., Citrus hystrix DC., Coriandrum sativum L., Cymbopogon citratus (DC.) Stapf, Eryngium foetidum L., Psophocarpus tetragonolobus (L.) DC., Senegalia pennata subsp. insuavis (Lace) Maslin, Seigler & Ebinger, and Solanum melongena ‘Kermit’ against NCDs. Despite having nothing in common , these plants have been selected based on their regular and frequent usages in most Thai cuisines. All plant samples were vetted for their origin and quality by the Department of Agriculture, Ministry of Agriculture and Cooperatives, Thailand. Thailand is renowned for its mouthwatering cuisine, consisting of a wide variety of textures and aromas emanating from local vegetable ingredients. Scientific reports have identified the health benefits of vegetables used in Thai dishes including antioxidant , anti-inAllium cepa Aggregatum Group (A. cepa), Allium fistulosum L. (A. fistulosum), Allium sativum L. (A. sativum), Citrus hystrix DC. (Ci. hystrix), Coriandrum sativum L. (Co. sativum), Cymbopogon citratus (DC.) Stapf (Cy. citratus), Eryngium foetidum L. (E. foetidum), Psophocarpus tetragonolobus (L.) DC. (P. tetragonolobus), Senegalia pennata subsp. insuavis (Lace) Maslin, Seigler & Ebinger (Se. pennata), and Solanum melongena ‘Kermit’ (So. melongena). Seven flavonoids were detected: quercetin, kaempferol, hesperidin, luteolin, apigenin, delphinidin, and cyanidin  contained two flavonoids at different concentrations. Four extracts possessed only one flavonoid: A. fistulosum (kaempferol), Cy. citratus (luteolin), E. foetidum (kaempferol), and Se. pennata (apigenin). Interestingly, no flavonoids were detected in A. sativum and So. melongena extracts. An aqueous ethanolic extract of Co. sativum was rich in quercetin (166.16 mg/100 g dry weight (DW)), followed by A. cepa and Ci. hystrix , while kaempferol (4.44–47.97 mg/100 g DW) was detected in A. fistulosum, Co. sativum, and E. foetidum extracts, with A. fistulosum extract exhibiting the highest (47.97 mg/100 g DW). Hesperdin (453.47 mg/100 g DW) was solely detected in Ci. hystrix extract, luteolin (4.57 mg/100 g DW) in Cy. citratus extract, apigenin (3.46 mg/100 g DW) in Se. pennata extract, and delphinidin (15.77 mg/100 g DW) in P. tetragonolobus extract. Cyanidin was also found in P. tetragonolobus (43.02 mg/100 g DW) and A. cepa (13.35 mg/100 g DW).High-performance liquid chromatography (HPLC) was employed to determine the specific phytochemical profiles covering the flavonoids and phenolic acids of vegetable extracts including cyanidin . Among tp-coumaric acid, and ferulic acid were detected. Caffeic acid and p-coumaric acid were general phenolics as they were observed in six extracts with different concentrations. The highest content of caffeic acid (246.99 mg/100 g DW) was detected in So. melongena extract that also contained minute amounts of p-coumaric acid (2.16 mg/100 g DW). The highest content of p-coumaric acid (68.13 mg/100 g DW) was detected in Cy. citratus extract, which also possessed the highest content of ferulic acid (123.34 mg/100 g DW) and marginal amounts of caffeic acid (15.65 mg/100 g DW). The second most abundant caffeic acid (52.69 mg/100 g DW) was detected in E. foetidum extract, which also contained minute amounts of p-coumaric acid and ferulic acid . An aqueous ethanolic extract of P. tetragonolobus contained the most varieties of phenolic acids including caffeic acid (16.13 mg/100 g DW), vanillic acid (15.71 mg/100 g DW), 4-hydroxybenzoic acid (10.80 mg/100 g DW), and p-coumaric acid (3.85 mg/100 g DW), while 4-hydroxybenzoic acid was only detected in P. tetragonolobus extract. Other than being observed in P. tetragonolobus extract, vanillic acid was also found in Co. sativum but in lower amounts (3.73 mg/100 g DW). This extract also contained caffeic acid (23.81 mg/100 g DW) and p-coumaric acid (5.20 mg/100 g DW). An aqueous ethanolic extract of A. fistulosum was found to possess ferulic acid (23.13 mg/100 g DW) and p-coumaric acid (7.12 mg/100 g DW), while Se. pennata contained only one phenolic acid: caffeic acid (14.92 mg/100 g DW). Interestingly, no phenolic acids were observed in A. cepa, A. sativum, and Ci. hystrix extracts.For phenolic acid determination , five phSe. pennata exhibited the highest TPC, followed by Ci. hystrix, So. melongena, Cy. citratus, A. fistulosum, P. tetragonolobus, E. foetidum, A. cepa, and Co. sativum, respectively, while A. sativum exhibited the lowest (twelve times lower than Se. pennata). A spectrophotometric analysis indicated that TPCs of all vegetable extracts ranged from 1.23 to 15.33 mg gallic acid equivalent (GAE)/g DW . The aquSe. pennata extract exhibiting the highest DPPH radical scavenging activity and A. sativum extract the lowest. For the FRAP assay, results were consistent with the DPPH radical scavenging activities. Se. pennata exhibited the highest reducing ability (62.33 μmol TE/g DW) of ferric iron (Fe3+) to ferrous iron (Fe2+), while A. sativum exhibited the lowest reducing activity (3.25 μmol TE/g DW). On the other hand, Ci. hystrix and So. melongena possessed the highest ORAC activities (415.92–418.32 μmol TE/g DW), while A. cepa exhibited the lowest (15.12 μmol TE/g DW) at approximately 27-fold lower than Ci. hystrix and So. melongena. Phenolics contribute to a wide range of health benefits, including being antioxidants. Antioxidant activities were investigated covering both hydrogen atom transfer (HAT) and single electron transfer (SET) mechanisms . The ferAll the vegetable extracts contained phenolic acids and flavonoids that contributed to their therapeutic potential, specifically against some NCDs. Therefore, all the extracts were tested for their therapeutic potential against critical enzymes involved in NCDs and non-enzymatic reactions involving anti-glycation properties. Ci. hystrix exhibited the highest lipase inhibition, while A. cepa exhibited the lowest.Lipase is a lipid-degradation enzyme, and lipase inhibitors prevent fatty acid accumulation as one characteristic of obesity . The resCo. sativum (58.4%), E. foetidum (31.2%), and Ci. hystrix (26.6%) showed inhibitory activities of more than 25%  converts angiotensin I to angiotensin II, leading to vasoconstriction and increased blood pressure ; therebyA. sativum, inhibited AChE activities in the range of 8.3–58.6% using extract concentration of 1 mg/mL , one type of dementia, have been attributed to (i) degradation of the neurotransmitter acetylcholine by acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), or (ii) accumulation of amyloid plaque formed during amyloidogenesis by β-secretase (BACE-1) . Therefo 1 mg/mL . Among tntration . The hig 1 mg/mL . Among tSe. pennata provided the most potent anti-glycation activities induced by both D-glucose and MG, while A. sativum and A. cepa exhibited poor ability to inhibit glycation reactions.The glycation reaction is a non-enzymatic reaction involving interaction between monosaccharides and amino acids, lipids, or nucleotides. Methylglyoxal (MG), as a by-product of glycolysis, can act as a potent protein-glycation inducing agent. Finally, glycation induced by either D-glucose or MG as advanced glycation end products (AGEs) contributed to several ailments including premature aging and diabetes . The resRelationships between vegetable extracts and TPCs, antioxidant activities, enzyme inhibitory activities and anti-glycation properties were investigated using principal component analysis (PCA) and hierarchical cluster analysis (HCA) to determine the particular characteristics of each vegetable extract. Information gained from these statistical analyses will be helpful to classify Thai vegetables according to their unique health-promoting characteristics. PCA results showed that TPCs, antioxidant activities, enzyme inhibitory activities, and anti-glycation properties of Thai vegetable extracts could be easily classified. . A. cepa and A. sativum (blue letters) were separated from others (red letters). A. cepa and A. sativum (Cluster 1) were separated from the others (cluster 2) and located far away from the centroid , while cluster 2 consisted of A. fistulosum, Ci. hystrix, Co. sativum, Cy. citratus, E. foetidum, P. tetragonolobus, Se. pennata, and So. melongena (red color). The outcome of HCA supported the PCA results , Allium fistulosum L. (A. fistulosum), Allium sativum L. (A. sativum), Citrus hystrix DC. (Ci. hystrix), Coriandrum sativum L. (Co. sativum), Cymbopogon citratus (DC.) Stapf (Cy. citratus), Eryngium foetidum L. (E. foetidum), Psophocarpus tetragonolobus (L.) DC. (P. tetragonolobus), Senegalia pennata subsp. insuavis (Lace) Maslin, Seigler & Ebinger (Se. pennata), and Solanum melongena ‘Kermit’ (So. melongena) were comparatively analyzed regarding their phenolic profiles (phenolic acids and flavonoids) and in vitro inhibitory activities against some NCDs. The health-promoting activities involved the inhibition of the key enzymes that control NCDs including lipase (obesity), α-amylase, and α-glucosidase (diabetes), angiotensin-converting enzyme (hypertension) and acetylcholinesterase, butyrylcholinesterase, and β-secretase (Alzheimer’s disease) as well as the non-enzymatic anti-glycation reaction (premature aging). Results showed that among these ten plant extracts, A. cepa exhibited the strongest angiotensin-converting enzyme (ACE) inhibition, while A. fistulosum could effectively fight against β-secretase (BACE-1). Nevertheless, A. sativum seemed to be the least active extract against these NCDs-related enzymes, in which no inhibitory activities against α–glucosidase, acetylcholinesterase (AChE), and BACE-1 were observed. However, high ACE inhibitory activity was observed in this plant extract, while other enzyme inhibitory activities were quite low (less than 50% inhibition). On the other hand, Ci. hystrix was the most active extract, which exhibited the strongest antioxidant activity and inhibitory activities against lipase, ACE and butyrylcholinesterase (BChE). Co. sativum was highly effective against α-amylase, while P. tetragonolobus exhibited the highest α-glucosidase inhibitory activity. Cy. citratus seemed to exhibit low to moderate enzyme inhibitory activities, with the exception of ACE inhibitory activity, which was more than 50% inhibition. E. foetidum exhibited the highest AChE inhibitory activity, while Se. pennata exhibited strong antioxidant activity and ACE inhibitory activity. Similarly, So. melongena was also a good source of antioxidants. From these results, it is of interest to group these plants according to their bioactivities and discuss the following topics in more detail; (i) Se. pennata exhibited the highest reducing and free radical scavenging abilities, while Ci. hystrix and So. melongena possessed the highest oxygen radical absorbance capacity; (ii) Ci. hystrix exhibited the highest lipase inhibition; (iii) Co. sativum and P. tetragonolobus were potential anti-diabetic agents with high inhibitions against carbohydrate-degrading enzymes; (iv) A. cepa, Ci. hystrix, and Se. pennata had the three highest ACE inhibitory activities; (v) Ci. hystrix and E. foetidum were potential anti-Alzheimer’s disease (AD) agents with high inhibitions against acetylcholine degrading enzymes, whereas A. fistulosum acted against amyloid generating enzyme, and (vi) Se. pennata was a potential anti-glycation agent.Interest in indigenous plants for their health benefits in terms of disease prevention beyond nutritional benefits is increasing. Thai cuisine consists of various spices and herbs with unique aromas and flavors that also have health-promoting bioactivities. Copious literature exists on plant beneficial health characteristics, but control of plant origins is lacking, leading to the absence of a comparative analysis of these vegetables. Here, ten vegetables used in Thai cuisine including Se. pennata, Ci. Hystrix, and So. melongena exhibited high antioxidant capacities with the highest total phenolic contents (TPCs). Our results concurred with previous literature suggesting that TPCs and antioxidant activities in various plant extracts were strongly correlated [Se. pennata (or Cha-om in Thai) exhibit strong odor (some define as stinky) and are normally consumed as a blanched vegetable or mixed in an omelet and eaten with spicy sauce. In our study, Se. pennata exhibited high ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities, suggesting that antioxidants in this vegetable extract likely possess an ability to transfer one electron to any potential electron acceptors. A previous report suggested that methanolic extract of Se. pennata leaves exhibited high TPCs of 45.3 µg gallic acid equivalent (GAE)/mg dry extract with half maximal effective concentration (EC50) of 3.6 mg extract/mg DPPH [50) of 2.0 mg/mL [Ci. hystrix and So. melongena exhibited high oxygen radical absorbance capacity (ORAC), suggesting that the antioxidants in these vegetable extracts likely donate hydrogen atoms to free radicals. The fruit peel of Ci. hystrix  is commonly used in many Thai recipes (such as chili paste and curry) for its strong and unique aroma. Eggplants in Thailand can be classified into 22 species of Solanum, with 10 cultivars commercially available [So. melongena can be consumed as fresh or blanched vegetables and are the main ingredient in curry. Previous literature also reported high TPCs and antioxidant activities of these two vegetables. Ethanolic extracts of Ci. hystrix fruit peel were previously reported to exhibit TPCs of 0.32 mg GAE/mg extract and DPPH radical scavenging activity with IC50 of 0.09 mg/mL [So. melongena  exhibited DPPH radical scavenging activities of 17.52–46.13% [Among the vegetable extracts, rrelated ,28. Youn/mg DPPH , while i.0 mg/mL . Our stuvailable . Fruits 2–46.13% .Ci. hystrix but not on its anti-obesity property. Interestingly, in this study, fruit peel extract of Ci. hystrix was found to exhibit the highest lipase inhibition, an activity that retards lipid absorption and, thus, is related to the control of obesity. As the most abundantly found flavonoid in fruit peel of Ci. hystrix, hesperidin exhibited IC50 activity of 52.4 µM against porcine pancreatic lipase [50 of 6.1 µM [50 of 4 µM [Ci. hystrix, it was previously found that phenolic acids were generally less active against lipase inhibition than flavonoids [Ci. hystrix with the highest content of hesperidin and moderate amounts of quercetin possessed a potential bioactivity against lipase. Many authors have reported on the biological activities of c lipase , while af 6.1 µM . Compare of 4 µM , quercetavonoids ,35. TherCo. sativum and P. tetragonolobus effectively inhibited α-amylase and α-glucosidase, respectively. Leaves and young shoots of Co. sativum (coriander) are added to many Thai dishes including soup or stir fry meat as a decorated vegetable with a strong unique aroma. However, recently, coriander has become popular consumed as a fresh side dish vegetable along with other main dishes. Oral administration of the ethanolic leaf extract to mice under induced insulin deficiency resulted in lowered blood glucose [50 value 2.5-times lower than acarbose, a synthetic anti-diabetic drug [Co. sativum leaves inhibited α-amylase with 19% inhibition using extract concentration of 1 mg/mL [50 of 14.60 µM against pancreatic α-amylase, the most abundant flavonoid in Co. sativum leaves as quercetin exhibited IC50 of 12.7 µM [50 of 20.4 µM [Co. sativum is a good candidate as an anti-diabetic agent, even its seeds [P. tetragonolobus (or winged bean) is a Thai local vegetable normally consumed as boiled or blanched young bean pod with many spicy sauces. At present, no report on young bean pod of P. tetragonolobus regarding its anti-diabetic property is available. Nevertheless, major phenolics in P. tetragonolobus such as cyanidin and delphinidin effectively inhibited Saccharomyces cerevisiae α-glucosidase with IC50 values of 17.0 and 4.1 µM, respectively, compared to acarbose with IC50 of 0.53 µM [P. tetragonolobus also contained moderate contents of phenolic acids including 4-hydroxybenzoic acid, vanillic acid, caffeic acid and p-coumaric acid, while phenolic acids generally inhibited α-glucosidase with a lesser effect than flavonoids [As vegetable extracts with potential anti-diabetic properties,  glucose ,35,36. T 1 mg/mL . Compare 12.7 µM , while t 20.4 µM . With hits seeds ,40,41. L 0.53 µM . P. tetravonoids ,43,44. Cavonoids . A. cepa, Ci. hystrix and Se. pennata had the three highest ACE inhibitory activities. These results concurred with previous reports indicating that quercetin-rich onion skin extract decreased ambulatory blood pressure in patients under metabolic syndrome (overweight/obese/hypertension) [A. cepa as quercetin exhibited IC50 of 43 µM against rabbit lung ACE [Ci. hystrix, exhibited half inhibitory activity of quercetin using a concentration of 0.5 mM [Se. pennata (but in low amounts compared to other vegetables) with IC50 of 5.7 mM [Interestingly, all vegetable extracts effectively inhibited ACE at more than 50% inhibition using extract concentration of 1 mg/mL. Among these, tension) . The prelung ACE , while hf 0.5 mM . Caffeicf 5.7 mM .Ci. hystrix and E. foetidum acted as inhibitors for acetylcholine-degrading enzymes, while A. fistulosum showed promise as a potential extract inhibiting amyloid production. Ci. hystrix and E. foetidum were rich in hesperidin and caffeic acid, respectively, while the consumption of hespiridin-rich extract or caffeic acid avoided cognitive dysfunction and learning deficit in vivo [50 against AChE and BChE at 22.8 and 48.9 µM, respectively, and caffeic acid at 23.36 and 29.19 µM, respectively [E. foetidum, usually used in the famous sour soup called “Tom Yum” was available; thus, future studies on the anti-AD properties of the caffeic acid rich extract of E. foetidum are required. When considering anti-AD via the inhibition of amyloid production, all extracts displayed mild to low BACE-1 inhibition. A. fistulosum and A. cepa gave the two highest anti-BACE-1 activities, albeit carrying different phytochemicals. A. cepa showed high quercetin, while A. fistulosum was high in kaempferol. Quercetin and kaempferol are well-known flavonoids exhibiting anti-BACE-1 properties with IC50 values at 5.4 and 14.7 µM, respectively [Co. sativum also possessed high quercetin (2.5-fold higher than A. cepa); however, the anti-BACE-1 activity was lower, indicating that multi-interaction of phytochemicals within the extract may display antagonist effects. For anti-AD properties, our data showed that using an extract concentration of 1 mg/mL,  in vivo ,49. Hespectively . Thus, hectively . Leaves Se. pennata may be a potential anti-glycation agent for both reactions. Se. pennata is typically fried with egg or with “Tom Yum” soup. The glycation reaction and its AGEs relate to free radical productions [Se. pennata exhibited high antioxidant activity covering the SET mechanism due to its high phenolics content. It remains unclear which phytochemicals in Se. pennata exhibit this property because only trace amounts of caffeic acid and apigenin were observed, even though these two compounds were documented for their anti-glycation properties [The glycation reaction leads to the formation of advanced glycation end products (AGEs) that contribute to diseases such as diabetes, AD, and premature aging . The reaductions . Hence, operties ,55.Ci. hystrix). However, ACE inhibitory activity was different from other bioactivities since it was located in a different axis (PC3). Moreover, ACE inhibitory activities of all vegetable extracts were higher than 50%, even though the extract concentration used in this enzyme inhibitory assay was lower (0.2 mg/mL) than others (1 mg/mL in other enzyme inhibitory assays and 0.63 mg/mL in glycation reactions). It was previously reported that other than phenolics that could act as ACE inhibitors, small peptides could also act as effective ACE inhibitors as well [A. cepa and A. sativum exhibited low TPCs, antioxidant activities, and anti-glycation properties. Cluster 2 consisted of A. fistulosum, Ci. hystrix, Co. sativum, Cy. citratus, E. foetidum, P. tetragonolobus, Se. pennata, and So. melongena exhibited high TPCs, antioxidant activities, and enzyme inhibitory activities. Additionally, the principal component analysis (PCA) suggested that the activities that lied in the same axis were closely related to each other. For example, TPCs, antioxidant activities, α-glucosidase inhibitory activities, AChE inhibitory activities, and anti-glycation properties were on the same axis (PC1). The extract with relatively high TPCs and antioxidant activities would potentially exhibit relatively high anti-α-glucosidase, AChE, or glycation activities as well , Allium fistulosum L. (A. fistulosum), Allium sativum L. (A. sativum), Citrus hystrix DC. (Ci. hystrix), Coriandrum sativum L. (Co. sativum), Cymbopogon citratus (DC.) Stapf (Cy. citratus), Eryngium foetidum L. (E. foetidum), Psophocarpus tetragonolobus (L.) DC. (P. tetragonolobus), Senegalia pennata subsp. insuavis (Lace) Maslin, Seigler & Ebinger (Se. pennata), and Solanum melongena ‘Kermit’ (So. melongena) were collected following the recommendation of the Department of Agriculture, Ministry of Agriculture and Cooperatives, Thailand. The samples were deposited at the Bangkok Herbarium (BK), Bangkok, Thailand. Physical appearance of the edible part, harvesting time and the herbarium voucher specimen are provided in A. cepa (bulps), A. fistulosum (leaves), A. sativum (bulps), Ci. hystrix (fruit peel), Co. sativum (leaves), Cy. citratus , E. foetidum (leaves), P. tetragonolobus (whole fruits), Se. pennata (young leaves), and So. melongena (whole fruits) were cleaned with deionized (DI) water before freeze-drying using a Heto PowerDry PL9000 Freeze Dryer  for 3 days. The dry samples were then ground into fine powder using a Philips 600W Grinder  and expressed as CIELAB units (L* represented dark (0) to white (100), a* represented green (−) to red (+), while b* represented blue (−) to yellow (+)) as shown in Ten vegetables including v/v) aqueous ethanol (1:10 ratio) at 37 °C for two hours. The mixture was centrifuged at 3800 g for 15 min using a Hettich® ROTINA 38R centrifuge . The supernatant was collected, while the residue was repeatedly extracted with the same procedure twice. The supernatants from three extractions were pooled, and ethanol was removed by a rotary evaporator . The dried extracts were re-dissolved in DMSO, filtered through a 0.45 µM polytetrafluoroethylene (PTFE) membrane syringe filter, and kept at −20 °C until analysis. The dry samples were extracted using 80% (v/v) aqueous methanol (40 mL), 6 N HCl (10 mL) and 0.5 g/L tert-butylhydroquinone (tBHQ). Prior to injection into the HPLC system, the extract (10 mg/mL) was filtered through a 0.22 μM PTFE membrane. Milli-Q water (18.2 MΩ.cm resistivity at 25 °C), HPLC-grade methanol, and HPLC-grade acetonitrile containing 0.05% (v/v) trifluoroacetic (TFA) were used as gradient mobile phases with a constant flow rate of 0.6 mL/min [p-coumaric acid , sinapic acid , and syringic acid (>97.0% T) were received from Tokyo Chemical Industry , while vanillic acid (≥97% HPLC) and gallic acid (97.5–102.5% T) were received from Sigma-Aldrich . The authentic flavonoid standards including quercetin , kaempferol (>97.0% HPLC), luteolin (>98.0% HPLC), hesperidin , naringenin , myricetin (>97.0% HPLC), and apigenin (>98.0% HPLC) were obtained from Tokyo Chemical Industry , while isorhamnetin (≥99.0% HPLC), cyanidin (≥96.0% HPLC), and delphinidin (≥97.0% HPLC) were from Extrasynthese . The phenolic acids were detected at 280 nm and 325 nm, while flavonoids were detected at 338 nm and 368 nm. HPLC chromatograms were shown in To determine the phenolic profile, high-performance liquid chromatography (HPLC) was employed using an Agilent 1100 HPLC system equipped with a photodiode array detector and a Zorbax Eclipse XDB–C18 column  as previously described . In brie6 mL/min . The auty is an area under the peak, a is a y-intercept, b is a slope of the calibration curve, and x is a standard concentration. LOD and LOQ were calculated using the following equation:aS is a standard deviation of the response (y-intercept), and b is a slope of the calibration curve. The intra-day precision was presented as a percentage of the relative standard deviation (%RSD) and calculated using the following equation:tRS is a standard deviation of the retention time, and tRMean is the mean of the retention time measured at all concentrations of each standard.Linear range, linear regression, correlation coefficients, limit of quantitation (LOQ), limit of detection (LOD), and relative standard deviation (RSD) of the standards were analyzed according to the protocol of Srinuanchai et al. 2019  as shownTotal phenolic contents (TPCs) were investigated using Folin’s phenol reagent as formerly described . Gallic Antioxidant activities of the extracts were determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) together with ferric reducing antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC) assays as previously described . Trolox,TM HT 96-well UV-visible microplate reader and Gen5 data analysis software . To determine the enzyme inhibitory activities of the extracts against some NCDs, key enzymes that control obesity (lipase), diabetes (α-amylase and α-glucosidase), hypertension (angiotensin-converting enzyme), and Alzheimer’s disease  were chosen for inhibitory reactions using the well-established protocols as previously described ,12,13,22Candida rugosa lipase , 50 µL of 0.2 mM 5-5′-dithiobis, 10 µL of 16 mM 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB) and 40 µL of the extract (5 mg/mL). The inhibitory activity was visualized as a decline in enzyme kinetics at 412 nm.Briefly, the lipase inhibitory reaction consisted of 100 µL of 0.01 mg/mL p-nitrophenyl-α-D-maltopentaoside and 50 µL of the extract (4 mg/mL), while the α-glucosidase inhibitory reaction consisted of 100 µL of 0.1 U/mL Saccharomyces cerevisiae α-glucosidase , 50 µL of 2 mM p-nitrophenyl-α-D-glucopyranoside and 50 µL of the extract (4 mg/mL). The inhibitory activity was visualized as a decline in enzyme kinetics at 405 nm.The α-amylase inhibitory reaction consisted of 100 µL of 30 mg/mL porcine pancreatic α-amylase , 50 µL of 30 mM o-phthaldialdehyde and 50 µL of the extract (0.4 mg/mL). The inhibitory activity was evaluated using an excitation wavelength of 360 nm and an emission wavelength of 485 nm as an end-point assay.The angiotensin-converting enzyme (ACE) inhibitory reaction consisted of 3 µL of 0.5 U/mL rabbit lung ACE (≥2 unit/mg), 30 µL of 3 mM hippuryl-histidyl-leucine, 15 µL of 20 mg/mL Electrophorus electricus AChE (1000 units/mg), 40 μL of 0.8 mM acetylthiocholine, 10 µL of 16 mM DTNB and 40 µL of the extract (5 mg/mL), while the butyrylcholinesterase (BChE) inhibitory reaction consisted of 100 µL of 0.5 µg/mL equine serum BChE (≥10 units/mg), 40 µL of 0.4 mM butyrylthiocholine, 10 µL of 16 mM DTNB and 40 µL of the extract (5 mg/mL). The inhibitory activity was visualized as a decline in enzyme kinetics at 412 nm. The β-secretase (BACE-1) inhibitory reaction was studied using a BACE-1 fluorescence resonance energy transfer (FRET) assay kit  according to the manufacturer’s recommendations. The inhibitory activity of the extract (20 µL of 5 mg/mL) was evaluated using an excitation wavelength of 320 nm and an emission wavelength of 405 nm as an end-point assay. The acetylcholinesterase (AChE) inhibitory reaction consisted of 100 μL of 20 ng The anti-glycation reaction induced by D-glucose consisted of 50 µL of 20 mg/mL bovine serum albumin  in 100 mM potassium phosphate buffer (pH 7.4) containing 0.02% (w/v) sodium azide, 25 µL of 1 M D-glucose and 25 µL of extract (2.52 mg/mL). For the anti-glycation reaction induced by methylglyoxal (MG), 25 µL of 4 mM MG was used instead of D-glucose. The reaction mixture was incubated at 37 °C for 2 weeks in the dark. The inhibitory activity was evaluated using an excitation wavelength of 330 nm and an emission wavelength of 410 nm as an end-point assay.A is the initial velocity of the control reaction with enzyme (control), a is the initial velocity of the control reaction without enzyme (control blank), B is the initial velocity of the enzyme reaction with extract (sample) and b is the initial velocity of the reaction with extract but without enzyme (sample blank). The percentage of enzyme inhibition using an end-point assay and anti-glycation reaction was evaluated using the same equation but changing from initial velocity to absorbance at a particular wavelength.The percentage of enzyme inhibition using enzyme kinetics was calculated using the following equation:®  to create a biplot. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) of TPCs, antioxidant activities and enzymatic and non-enzymatic inhibitory activities of Thai vegetable extracts were performed using XLSTATn = 3), with data expressed as mean ± standard deviation (SD). Statistical analysis was performed using the statistical package for the social sciences . Significant difference at p < 0.05 of more than two data was calculated using one–way analysis of variance (ANOVA), followed by Duncan’s multiple comparison test, while significantly difference at p < 0.05 of two data was calculated by Student’s t-test. All experiments were evaluated in triplicate (