A Study About The Protective Effect Of Alliin On Injury Of Human Umbilical Vein Endothelial Cells Induced By Ox-LDL

Objective:To investigate the protective effect of Alliin on oxidized low-density lipoprotein(ox-LDL)induced injury of human umbilical vein endothelial cells(HUVEC),and to study the related proteins and signaling pathways involved in this protective mechanism.Methods:80mg/Lox-LDL induced HUVEC for 24 hours to establish a HUE VC injury model.Total cholesterol(TC)and free cholesterol(Fch)kits were used to detectTC and Fch content;CCK8 kit was used to detect different concentrations and different time about the cell viablitity of HUVEC;the effect of Alliin on the expression of cystathionine y lyase(CSE)in the HUE VC injury model was detected by Western blotting,and the HS-content in HUVEC supernatant was detected by methylene blue method.Quantitative proteomics was used to quantitatively analyze the differential proteins in cell samples from the blank control group(Con),HUVEC injury model group(Mod),and Alliin group(Alliin);among the quantified proteins,1.5 times for differential expression as the threshold of change,t-test P-vaule<0.05 was used as the significance threshold for statistical analysis of differential proteins in Alliin/Con,Alliin/Mod,and Mod/Con;using GO enrichment,KEGG pathway enrichment,and KEGG pathway enrichment clustering performed functional enrichment analysis of the differential proteins in Alliin/Mod,and performed Western blotting verificationon the significantly enriched KEGG pathway.Result:80mg/L ox-LDL induced a large number of red-stained granules in HUVEC cells within 24 hours,and the Fch content exceeded 50%of TC content,and the HUVEC injury model was successfully established;12.5～50mg/L Alliin could significantly promote the cell viability of HUVEC injury model(P<0.05);Alliin can significantly increase the expression of CSE protein in HUVEC injury model(P<0.05),and 50mg/L Alliin can significantly increase the HS-content of HUVEC injury model cells(P<0.05);Quantitative proteomics results showed that a total of 6173 proteins were identified,of which 5162 were quantifiable;differential protein statistics showed that a total of 108 proteins were up-regulated and 116 proteins were down-regulated in Alliin/Con(P<0.05);A total of 33 proteins were up-regulated in Mod,17 proteins were down-regulated in Alliin/Mod(P<0.05);a total of 106 proteins were up-regulated,and 147 proteins were down-regulated in Mod/Con(P<0.05);Alliin/Mod differential protein functional enrichment and western Blotting verification results showed that Alliin can regulate the cholesterol metabolism pathway of HUVEC injury model by up-regulating the expression of LDLR(P<0.05),ApoC(P<0.05)and down-regulating the expression of ApoB(P<0.05).Conclusion:Alliin can protect the human umbilical vein endothelial cells from injury induced by ox-LDL by regulating the CSE/H2S pathway and the cholesterol metabolism pathway.