川芎嗪对脂多糖诱导内皮细胞凋亡及Sirt1/FoxO1通路的影响

Effect of tetramethylpyrazine on apoptosis of endothelial cells induced by lipopolysaccharide and Sirt1/FoxO1 pathway

目的研究川芎嗪(tetramethylpyrazine,TMP)对脂多糖(LPS)诱导内皮细胞凋亡及沉默信息调节因子1/叉头转录因子O1(SIRT1/Fox O1)通路的影响。方法培养人脐静脉内皮细胞(HUVECs)并分为对照组、LPS组和TMP组。对照组用不含药物的DMEM处理,LPS组用含有10μg/ml LPS的DMEM处理,TMP组用含有含有10μg/ml LPS及不同浓度TMP(10-10,10-12,10-14mol/L)的DMEM处理,检测细胞活力、凋亡率、凋亡基因及Sirt1/Fox O1通路分子的表达。结果与对照组比较,LPS组HUVECs的细胞活力及Bcl-2、乙酰化-Fox O1的蛋白表达明显降低,凋亡率及Bax、Caspase-9、Caspase-3、Sirt1的蛋白表达量明显增加(P<0.05);与LPS组比较,10-10,10-12,10-14mol/L TMP组的细胞活力明显增加、凋亡率明显降低(P<0.05),且TMP浓度越高,细胞活力的增加及凋亡率的降低越明显(P<0.05);与LPS组比较,10-10mol/L TMP组的Bax、Caspase-9、Caspase-3、Sirt1的蛋白表达量明显增加,Bcl-2、乙酰化-Fox O1的蛋白表达量明显减少(P<0.05),Fox O1的蛋白表达量无明显变化(P>0.05)。结论TMP对LPS诱导内皮细胞凋亡具有抑制作用,且该作用可能与Sirt1/Fox O1通路的抑制有关。

Objective To study the effects of tetramethylpyrazine(TMP)on apoptosis of endothelial cells induced by lipopolysaccharide(LPS)and silent information regulator protein/forkhead box O1(SIRT1/Fox O1)pathway.Methods Human umbilical vein endothelial cells(HUVECs)were cultured and divided into control group,LPS group,and TMP group.The cells were cultured with DMEM without drug treatment in control group,DMEM containing 10μg/ml LPS in LPS group,and DMEM containing 10μg/ml LPS and different concentrations of TMP(10-10,10-12,10-14 mol/L)in TMP group.Cell viability,apoptotic rate,expression of apoptotic gene and Sirt1/Fox O1 pathway molecule were detected.Results Compared with control group,the cell viability and the protein expression of Bcl-2,acetylated-FoxO1 in LPS group significantly decreased,while the apoptotic rate and the expression of Bax,Caspase-9,Caspase-3 and Sirt1 significantly increased(P<0.05).Compared with LPS group,the cell viability was significantly increased,while the apoptotic rate significantly decreased in TMP groups in a concentration-dependent manner(P<0.05).Compared with LPS group,the expression of Bax,Caspase-9,Caspase-3 and Sirt1 was significantly increased,while the expression of Bcl-2 and acetylated Fox O1 was significantly decreased in 10-10 mol/L TMP group(P<0.05),and the expression of Fox O1 did not change significantly(P>0.05).Conclusion TMP can inhibit the apoptosis of endothelial cells induced by LPS,which may be related to the inhibition of Sirt1/Fox O1 pathway.