消渴平含药血清对高糖诱导大鼠肾小球系膜细胞凋亡的影响

Effects of Serum Containing Xiaokeping(消渴平)on Apoptosis of Rat’Mesangial Cells Induced by High Glucose

目的:观察消渴平(Xiaokeping,XKP)含药血清对高糖诱导的大鼠肾小球系膜细胞凋亡的影响,探讨其治疗DKD的作用机理。方法:使用不同浓度的消渴平含药血清及厄贝沙坦含药血清与高糖共同培养大鼠肾小球系膜细胞,应用TUNEL法检测细胞凋亡,荧光定量PCR检测大鼠肾小球系膜细胞中B淋巴细胞瘤基因相关X蛋白(Bax mRNA)、B淋巴细胞瘤基因-2(Bcl-2 mRNA)表达,Western-blot检测系膜细胞中活化半胱天冬酶3(cleaved caspase-3)、Bax、Bcl-2及细胞色素C(Cytochrome c,Cyt-C)的蛋白表达情况。结果:与空白对照组相比,高糖组细胞凋亡显著增加,cleaved caspase-3和Bax、Cyt-C表达显著增加(P<0.01),Bcl-2表达明显减少(P<0.01)。与高糖组相比,消渴平高剂量组可显著抑制高糖诱导的系膜细胞凋亡,抑制系膜细胞cleaved caspase-3和Bax、Cyt-C表达(P<0.05),增加Bcl-2表达(P<0.05),与厄贝沙坦组相比较无明显差异(P>0.05)。结论:消渴平通过抑制系膜细胞Bax表达,提高Bcl-2表达,减少细胞色素C易位,从而对高糖诱导的系膜细胞凋亡具有保护作用,这可能是其防治DKD的机制之一。

Objective:To observe the effects of Xiaokeping on the apoptosis of rat’s mesangial cells induced by high glucose,and to explore the mechanism of its prevention and treatment of diabetic kidney disease.Methods:Rat’mesangial cells were co-cultured with different concentrations of serum containing XKP or Irbesartan and high glucose.Cell apoptosis was detected by TUNEL,Bax and Bcl-2 mRNA expressions in mesangial cells of rat’s glomerulus were detected by fluorescence quantitative PCR,and protein expressions of cleaved caspase-3,Bax,Bcl-2 and cytochrome C in mesangial cells were detected by Western-blot.Results:Compared with the blank control group,cell apoptosis was significantly increased in the high-glucose group,expressions of cleaved caspase-3,Bax and cytochrome C were significantly increased(P<0.01),expression of Bcl-2 was significantly decreased(P<0.01).Compared with the high-glucose group,the high-dose group of XKP could significantly inhibit the high-glucose-induced mesangial cell apoptosis,inhibit the expressions of mesangial cells cleaved caspase-3,Bax and cytochrome C(P<0.05),increase the expression of Bcl-2(P<0.05),with no significant difference was founded compared with the Irbesartan group(P>0.05).Conclusion:XKP can significantly inhibit the expression of Bax in mesangial cells,improve the expression of Bcl-2,and reduce the translocation of cytochrome C,so as to have a protective effect on the apoptosis of mesangial cells induced by high glucose,which may be one of the mechanisms of its prevention and treatment of diabetic kidney disease.