| Objective:Different concentrations of EGCG were used to treat the endothelial cells on the surface of the scaffold material,and the concentrations of IL-6,TNF-α,ICAM-1 inflammatory factors and expressions of apoptosis-related genes such as Casepase-3,Casepase-8,Casepase-9,etc.were compared between groups Differences in levels,to explore the regulatory role of EGCG on endothelial cell proliferation and apoptosis,lay the foundation for the study of EGCG to prevent restenosis after vascular stent,and provide experimental basis for the clinical application of EGCG.Method:HUVEC cells were cultured on 316 L stainless steel sheet with 1640 medium containing 10% fetal bovine serum.Divided into HUVEC control group,316 L stainless steel sheet control group,12.5μmol / L EGCG,25μmol / L EGCG,50μmol / L EGCG and 100μmol / L EGCG experimental group.The cells and cell supernatant were collected after 24 hours,72 hours,and 120 hours.The MTT method was used to detect the survival rate of HUVEC cells;the ELISA method was used to determine the levels of IL-6,TNF-α and ICAM-1 in the cell supernatant;the Q-PCR method was used to determine the mRNA expression levels of apoptosis-related genes in the cells;Flow cytometry was used to measure the apoptosis.Statistical analysis was performed using IBM SPSS 18.0,the analysis of variance was used to compare the differences of various indicators between different drug administration groups,and the LSD method was used to compare each group pairwise;Non-normal data using rank sum test,with P <0.05 as the difference is statistically significant.Result:The proliferation rate of HUVEC cells in each dose group at 24 hours,72 hours,and 120 hours was significantly lower than that in the control group and 316 L stainless steel sheet control group(P<0.05).Seven days after EGCG administration,the ratios of early apoptosis in different dose groups were 67.99 ± 4.81%,70.60 ± 4.05%,62.68 ± 7.71%,67.34 ± 9.40%,which was lower than 86.69 ± 7.74% in blank group and 76.14 in 316 L group ± 0.66%;the ratios of late apoptosis in different dose groups were 3.97 ± 1.97%,3.17 ± 0.66%,3.46 ± 0.79%,2.52 ± 0.54%,except that the 100μmol / L EGCG group was lower than the blank control group.It was higher than 3.00 ± 4.16% of the blank control group,and all the administration groups were lower than 4.23 ± 0.66% of the 316 L control group.After 24 hours of EGCG administration,the concentration of IL-6 in the cell supernatant of each administration group was lower than that of the 316 L stainless steel iron control group(P<0.001);48 hours after administration,12.5 μmol/L and 50 μmol/L groups The IL-6 concentration of the cell supernatant was lower than that of the 316 L stainless steel iron sheet control group,but the IL-6 concentration of the cell supernatant of the other two drug administration groups was higher than the 316 L stainless steel iron sheet control group(P<0.001);administration for 72 hours After 96 hours and 120 hours,the concentration of IL-6 in the cell supernatant of each administration group was lower than that of the 316 L stainless steel iron control group(P<0.05).After 24 hours,48 hours and 72 hours of administration,the ICAM-1 concentration of the cell supernatant of each administration group was lower than that of the 316 L stainless steel iron control group(P<0.001);after 96 hours of administration,each administration group The concentration of ICAM-1 in the cell supernatant was lower than that in the control group of 316 L stainless steel iron tablets(P<0.05);120 hours after administration,the concentration of ICAM-1 in the cell supernatant except the 12.5μmol/L group was higher than that of 316 L stainless steel iron Except for the tablet control group,the concentration of ICAM-1 in the cell supernatant of the other three drug administration groups was lower than that of the 316 L stainless steel iron control group(P<0.05).After 24,48 and 72 hours of EGCG administration,the concentration of TNF-α in the cell supernatant of each administration group was lower than that of the 316 L stainless steel iron control group(P<0.001);The concentration of TNF-α in the cell supernatant of the μmol/L group was higher than that of the 316 L stainless steel iron sheet control group.>0.05);120 hours after administration,the concentration of TNF-α in the cell supernatant of each administration group was lower than that of the 316 L stainless steel iron control group(P<0.001).Seven days after EGCG administration,except for the 12.5mol / L EGCG group,the expression of casspase-3 gene was slightly lower than that of the 316 L stainless steel iron control group,and the expression of caspase-3 gene in the other three administration groups was higher than that of the 316 L stainless steel iron control group(P <0.05);the expression of caspase-8,caspase-9 and Fas genes in all administration groups was higher than that in the 316 L stainless steel iron control group(P <0.05).Seven days after the administration of EGCG,the expressions of Atg-5,Atg-7 and Atg-12 genes in all administration groups were higher than those in the 316 L stainless steel iron control group(P <0.05).Conclusion:EGCG can reduce the proliferation of endothelial cells on the surface of the stent,promote cell apoptosis,and reduce the concentration of cytokines;EGCG has an important role in preventing and controlling atherosclerosis after stent restenosis,and provides experimental evidence for the clinical application of EGCG. |
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