Smad3对高糖环境下足细胞凋亡的影响

发布时间：2018-11-17 13:11

【摘要】：目的探讨果蝇Mad基因3哺乳动物类似基因(Smad3)对高糖环境下足细胞凋亡的影响。方法足细胞分为对照组、高糖组、干扰组,对照组、高糖组在实验开始前转染siRNA control,干扰组转染Smad3 siRNA。高糖组、干扰组用含有25 mmol/L的D葡萄糖细胞培养液培养,对照组用含有5 mmol/L的D葡萄糖细胞培养液培养。Western印迹检测Smad3、磷酸化的Smad3(p-Smad3)、酶切的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)、B细胞淋巴瘤(Bcl)-2相关X蛋白(Bax)、Bcl/白血病-x基因长片段(Bcl-xl)水平,流式细胞术检测细胞凋亡。结果高糖组细胞中Smad3水平与对照组相比无明显差异(P0.05),而p-Smad3水平明显高于对照组(P0.05)。干扰组细胞中Smad3、p-Smad3水平均明显低于高糖组(P0.05)。高糖组细胞凋亡率明显高于对照组,干扰组明显低于高糖组(均P0.05)。高糖组Cleaved Caspase-3、Bax水平明显高于对照组,Bcl-xl水平明显低于对照组(均P0.05),干扰组Cleaved Caspase-3、Bax水平均明显低于高糖组,Bcl-xl水平明显高于高糖组(均P0.05)。结论高糖环境下,足细胞中Smad3磷酸化水平升高,细胞凋亡增多,而下调Smad3可以抑制高糖诱导的足细胞凋亡,抑制Cleaved Caspase-3、Bax表达,促进Bcl-xl表达。
[Abstract]:Objective to investigate the effect of Mad gene 3 mammalian analogous gene (Smad3) of Drosophila melanogaster on apoptosis of podocyte in high glucose environment. Methods podocytes were divided into three groups: control group, high glucose group, interference group, control group and high glucose group. Before the experiment, the siRNA control, interference group was transfected with Smad3 siRNA.. High glucose group, interference group were cultured with D glucose cell culture medium containing 25 mmol/L, while control group were cultured with D glucose cell culture medium containing 5 mmol/L. Smad3, phosphorylation Smad3 (p-Smad3) was detected by Western blot. The level of (Bcl) 2 associated X protein (Bax), Bcl/ leukaemia X gene long fragment (Bcl-xl) in Cleaved Caspase-3), B cell lymphoma was determined by enzyme digestion of cysteine-containing aspartate protease 3 (Bcl) 2). Apoptosis was detected by flow cytometry. Results there was no significant difference in the level of Smad3 between the high glucose group and the control group (P0.05), but the level of p-Smad3 was significantly higher than that of the control group (P0.05). The level of Smad3,p-Smad3 in interference group was significantly lower than that in high glucose group (P0.05). The apoptosis rate of high glucose group was significantly higher than that of control group, and that of interference group was significantly lower than that of high glucose group (P0.05). The levels of Cleaved Caspase-3,Bax and Bcl-xl in high glucose group were significantly higher than those in control group (P0.05), Cleaved Caspase-3,Bax level in interference group was significantly lower than that in high glucose group, and Bcl-xl level was significantly higher than that in high glucose group (P0.05). Conclusion the level of Smad3 phosphorylation in podocytes was increased and apoptosis was increased under high glucose condition. Down-regulation of Smad3 could inhibit the apoptosis of podocytes induced by high glucose, inhibit the expression of Cleaved Caspase-3,Bax and promote the expression of Bcl-xl.
【作者单位】：新疆医科大学第一附属医院VIP内科;新疆医科大学附属中医医院干一科;
【基金】：新疆维吾尔自治区科技攻关和重点科技项目(201233398)
【分类号】：R587.2

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