G-蛋白偶联胆汁酸受体对高糖诱导大鼠冠状动脉内皮细胞功能的影响

EFFECT OF TGR5 ON CORONARY ARTERY ENDOTHELIAL CELLS INDUCED BY HIGH GLUCOSE IN RATS

目的探讨G-蛋白偶联胆汁酸受体(TGR5)对高糖诱导大鼠心脏动脉内皮细胞粥样硬化的影响。方法提取清洁级雄性SD大鼠冠状动脉内皮细胞于高糖培养基内传代培养,分为对照组、空载组和TGR5转染组。空载组细胞接受空载质粒转染,TGR5转染组接受TGR5质粒转染和INT-777激活。MTT法测定和比较各组内皮细胞增殖,Annexin-V/PE双染法测定和比较各组内皮细胞凋亡。RT-PCR法检测和比较各组内皮细胞ICAM-1和VCAM-1 mRNA表达,Western blot法测定和比较各组内皮细胞ICAM-1和VCAM-1蛋白表达。测定和比较各组内皮细胞SOD活性、MDA、LDH和GSH-Px表达。结果 0h时,各组内皮细胞增殖率差异无统计学意义(P>0.05),12 h及24 h时,TGR5转染组细胞增殖速度显著高于对照组及空载组(P<0.05)。对照组细胞凋亡率33.76%及空载组细胞凋亡率27.77%显著高于TGR5转染组细胞凋亡率12.36%(P<0.05)。TGR5转染组ICAM-1和VCAM-1 mRNA表达均显著低于对照组和空载组(P <0.001),TGR5转染组ICAM-1和VCAM-1蛋白表达均显著低于对照组和空载组(P<0.001),TGR5转染组内皮细胞SOD活性和GSH-Px表达高于空载组和对照组内皮细胞(P<0.05),TGR5转染组内皮细胞LDH和MDA表达低于空载组和对照组内皮细胞(P<0.05)。结论激活TGR5能够有效阻止大鼠心脏冠状动脉内皮细胞免于高糖诱导性损伤,可能为临床防治心脏冠状动脉粥样硬化提供指导。

Objective To investigate the effect of G-protein coupled bile acid receptor(TGR5)on atherosclerosis in rat cardiac artery endothelial cells induced by high glucose.Methods Extraction of glucose and coronary artery endothelial cells male SD rats were cultured in culture medium,divided into control group,vector group and TGR5 transfection group.Vector group cells were transfected with empty plasmid,and TGR5 transfection group received TGR5 plasmid transfection and INT-777 activation.MTT method was used to measure and compare the proliferation of endothelial cells in each group.Annexin-V/PE double staining method was used to measure and compare the apoptosis of endothelial cells in each group.RT-PCR method was used to detect and compare the expression of ICAM-1 and mRNA VCAM-1 in endothelial cells of each group.Blot Western method was used to measure and compare the expression of ICAM-1 and VCAM-1 protein in endothelial cells of each group.The expression levels of SOD,MDA,LDH and GSH-Px in endothelial cells of each group were measured and compared.Results There was no significant difference in the proliferation rate of endothelial cells between groups at 0h(P>0.05),at 12 h and24 h,TGR5 transfection group’s proliferation rates were significantly higher than that of control group and vector group(P<0.05).The apoptosis rate of the control group(33.76%)and the apoptotic rate of the vector group(27.77%)were significantly higher than that of TGR5 transfection group(12.36%)(P<0.05).The expression levels of ICAM-1 and VCAM-1 mRNA of TGR5 transfection group were significantly lower than those of the control group and vector group(P<0.001),the expression levels of TGR5 and VCAM-1 protein of TGR5 transfection group were significantly less than those of the control group and vector group(P<0.001).The expression levels of SOD activity and GSH-Px of endothelial cells of TGR5 transfected group were significantly higher than those of the control group and vector group(P<0.05),the expression levels of LDH and MDA in endothelial cells of TGR5 transfection group were lower than those of the control group and empty group(P<0.05).Conclusion Activation of TGR5 can effectively prevent rat coronary artery endothelial cells from high glucose-induced injury,which may provide guidance for clinical prevention and treatment of coronary atherosclerosis.