四逆汤对同型半胱氨酸损伤的EAhy926细胞caveolin-1和eNOS表达的影响

Effect of Sini decoction on expression of caveolin-1 and eNOS in EAhy926 cells injured by homocysteine

目的:探讨四逆汤(Sini decoction,SND)防治内皮细胞损伤的机制及陷窝蛋白1(caveolin-1)和一氧化氮(NO)系统在其中的作用。方法:建立同型半胱氨酸(Hcy)损伤的人脐静脉内皮融合细胞(EAhy926细胞)模型,观察四逆汤的保护作用及其对NO系统和caveolin-1的影响。结果:Hcy加入后细胞生长缓慢,贴壁细胞数明显减少,NO浓度明显减低(P<0.05),caveolin-1 mRNA和蛋白表达明显增强(P<0.05),内皮型NO合酶(eNOS)mRNA和蛋白表达明显减弱(P<0.05);四逆汤处理组贴壁细胞及细胞形态明显改善,NO浓度较Hcy模型组明显升高(P<0.05),caveolin-1 mRNA和蛋白表达较Hcy模型组明显减弱(P<0.05),eNOS mRNA和蛋白表达较Hcy模型组明显增强(P<0.05),其中以四逆汤1.0 kg/L+Hcy 4.0μmol/L组最明显。结论:Hcy可能通过增加caveolin-1表达和抑制eNOS表达而损伤人脐静脉内皮细胞,而四逆汤通过抑制caveolin-1表达和增加eNOS的表达拮抗Hcy对细胞的损伤。

AIM：To investigate the mechanism of Sini decoction in treating human vascular endothelial cell injury and the roles of caveolin-1 and nitric oxide （NO） system in this procedure. METHODS：Model of human umbilical vein endothelial EAhy926 cells injured by homocysteine （Hcy） was established. The protective effect of Sini decoction on the injured EAhy926 cells was observed, and the expression of caveolin-1 and endothelial nitric oxide synthase （eNOS） was detected by real-time fluorescence quantitative PCR and Western blotting. RESULTS：Compared with control group, the Hcy-treated EAhy926 cells showed reduced adherent cell number and NO concentration in culture supernatant, decreased expression of eNOS mRNA and protein, and increased expression of caveolin-1 mRNA and protein （all P〈0.05）. Compared with Hcy group, better growth of adherent cells, elevated NO concentration in culture supernatant, attenuated expression of caveolin-1 mRNA and protein, and enhanced expression of eNOS mRNA and protein in Sini decoction groups were observed （all P〈0.05）. CONCLUSION：Homocysteine may injure EAhy926 cells by enhancing the expression of caveolin-1 and suppressing the expression of eNOS, while Sini decoction may protect EAhy926 cells by suppressing the expression of caveolin-1 and enhancing the expression of eNOS.