缬沙坦对高糖诱导大鼠肾小球系膜细胞活性损伤的干预作用

Intervention of valsartan on high glucose-induced injury of rat glomerular mesangial cells activity

目的探讨缬沙坦对高糖诱导的大鼠肾细胞增殖和凋亡的影响与机制。方法利用高糖诱导大鼠肾小球系膜细胞HBZY-1损伤,并给予不同浓度的缬沙坦。Western blot检测增殖凋亡相关蛋白细胞周期蛋白D1(Cyclin D1)、活化的天冬氨酸特异性半胱氨酸蛋白酶3(Cleaved caspase-3)、Bcl-2相关X蛋白(Bax)、B细胞淋巴瘤/白血病-2(Bcl-2)和NOTCH1信号通路蛋白jagged1、NOTCH1的表达,噻唑蓝(MTT)比色法测定细胞增殖活力,流式细胞仪评估细胞周期与凋亡。采用NOTCH1信号通路激活剂Jagged 1/Fc融合蛋白和缬沙坦处理高糖诱导的HBZY-1,观察其对细胞的增殖、周期和凋亡的影响。结果高糖诱导的HBZY-1中Cyclin D1、Bcl-2蛋白表达水平、24 h、48 h和72 h的细胞增殖活力、S期细胞比例显著降低,G0-G1期细胞比例、Cleaved caspase-3、Bax、jagged1和NOTCH1蛋白表达水平、细胞凋亡率明显升高(P<0.05)。0.01、0.1、1μmol/L缬沙坦显著提高Cyclin D1、Bcl-2蛋白表达水平、24 h、48 h、72 h的细胞增殖活力和S期细胞比例,明显降低G0-G1期细胞比例、Cleaved caspase-3、Bax、jagged1、NOTCH1蛋白表达水平与细胞凋亡率,且均呈浓度依赖性(P <0.05)。NOTCH1信号通路激活剂Jagged 1/Fc融合蛋白部分逆转缬沙坦对高糖处理的HBZY-1增殖、周期和凋亡的影响。结论缬沙坦通过抑制NOTCH1信号通路,促进高糖处理的大鼠肾小球系膜细胞增殖与周期,并减轻细胞凋亡。

Objective To investigate the effect and mechanism of valsartan on high glucose-induced rat kidney cell proliferation and apoptosis.Methods High glucose was used to induce HBZY-1 damage in rat mesangial cells,and different concentrations of valsartan were given.Western blot detected of proliferation and apoptosis-related proteins Cyclin D1,Cleaved caspase-3,Bcl-2 related X protein(Bax),B cell lymphoma/leukemia-2(Bcl-2),and NOTCH1 signaling pathway protein jagged1,NOTCH1 expression,MTT colorimetric method was performed to determine cell proliferation activity,and flow cytometry was applied to evaluate cell cycle and apoptosis.The high glucose-induced HBZY-1 was treated with NOTCH1 signaling pathway activators Jagged 1/Fc fusion protein and valsartan,and its effects on cell proliferation,cell cycle and apoptosis were observed.Results The expression level of Cyclin D1,Bcl-2 protein,cell proliferation activity at 24 h,48 h and 72 h,and S-phase cell ratio in HBZY-1 induced by high glucose were significantly decreased,and G0-G1-phase cell ratio,Cleaved caspase-3,Bax,jagged1 and NOTCH1 protein expression levels and apoptosis rate were greatly increased(P<0.05).0.01,0.1,1μmol/L valsartan obviously improved the expression levels of Cyclin D1,Bcl-2,cell proliferation activity at 24 h,48 h and 72 h,and S-phase cell ratio,while dramatically reduced G0-G1-phase cell ratio,the expression levels of Cleaved caspase-3,Bax,jagged1,NOTCH1 and the apoptosis rate,all in a concentration-dependent manner(P<0.05).NOTCH1 signaling pathway activator Jagged 1/Fc fusion protein partially reversed the effects of valsartan on the proliferation,cell cycle and apoptosis of HBZY-1 treated with high glucose.Conclusions Valsartan can promote the proliferation and cell cycle of rat mesangial cells treated with high glucose by inhibiting the NOTCH1 signaling pathway,and reduce the apoptosis.