[沉默信息调节剂7对高糖环境下小鼠肾足细胞增殖和凋亡的影响]。

Min Feng, Ting Lin, Xia-Xia Chen, Xiao-Ling Yang, Qi Lyu, Jun-Ping Wen
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引用次数: 0

摘要

目的:探讨高糖环境下沉默信息调节因子7(SIRT7)对小鼠肾足细胞增殖和凋亡的影响及其机制。方法:将高糖培养不同处理方法的小鼠肾足细胞分为对照组(control)、高糖组(HG)、高糖+转染SIRT7过表达载体(pcDNA3.1-SIRT7)组(SIRT7 OE+HG)、高糖+转染阴性对照载体(pcDNA3.1)组(SIRT7 OE- nc +HG)、高糖+转染小干扰RNA-SIRT7 (siRNA-SIRT7)组(siRNA-SIRT7+HG)、高糖+转染siRNA-SIRT7对照组(siRNA-SIRT7- nc + HG)。CCK-8法检测细胞增殖活力。流式细胞术检测细胞凋亡率。采用qRT-PCR检测SIRT7 mRNA表达水平。Western blot检测Nephrin蛋白表达及Wnt/β-catenin信号通路关键因子的表达。结果:CCK-8结果显示,与对照组相比,HG组小鼠肾足细胞增殖活性降低(P<0.05)。转染SIRT7过表达载体或小干扰RNA-SIRT7后,与HG组相比,siRNA-SIRT7组细胞增殖活性进一步降低(P<0.05), SIRT7 OE+HG组细胞增殖活性增强(P<0.05)。流式细胞术结果显示,与对照组相比,HG组细胞凋亡率升高(P<0.05)。与HG组比较,siRNA SIRT7+HG组细胞凋亡率显著升高(P<0.05), SIRT7 OE+HG组细胞凋亡率显著降低(P<0.05)。与对照组相比,HG组Nephrin、Wnt5a、β-catenin的表达均受到抑制(P<0.05)。与HG组相比,siRNA-SIRT7组可下调Nephrin、Wnt5a、β-catenin的表达水平(P<0.05), SIRT7过表达可上调SIRT7 OE+HG组Nephrin、Wnt5a、β-catenin的表达水平(P<0.05)。结论:高糖环境是抑制小鼠肾足细胞增殖和诱导细胞凋亡的重要因素。SIRT7过表达可激活Wnt/β-catenin信号通路,上调β-catenin表达,从而逆转上述效应。
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[Effects of silence information regulator 7 on proliferation and apoptosis of mouse renal podocytes under high glucose environment].

Objective: To investigate the effects and its mechanisms of silence information regulator 7(SIRT7)on mouse renal podocytes proliferation and apoptosis under high glucose environment. Methods: Mouse renal podocytes cultured with high glucose and treated with different methods were divided into the following groups:control group(Control),high glucose group(HG),high glucose+transfecting with SIRT7 overexpression vetor(pcDNA3.1-SIRT7) group(SIRT7 OE+HG),high glucose+transfecting with the negative control vetor(pcDNA3.1)group(SIRT7 OE-NC+HG),high glucose+transfecting with small interfering RNA-SIRT7 (siRNA-SIRT7) group (siRNA-SIRT7+HG), high glucose+ transfecting with siRNA-SIRT7 control group (siRNA-SIRT7-NC+ HG). Viability of proliferation was examined by CCK-8 method.Rate of apoptosis was detected by flow cytometry. The level of SIRT7 mRNA expression was measured by qRT-PCR. Western blot was performed to detect the protein expression of Nephrin and key factors of Wnt/β-catenin signaling pathway. Results: The CCK-8 result showed that,compared with control group, the proliferative activity of mouse renal podocytes in HG group was decreased (P<0.05). After transfected with SIRT7 overexpression vetor or small interfering RNA-SIRT7,compared to HG group,the cell proliferation activity was further decreased in siRNA-SIRT7 group(P<0.05),but it was enhanced in SIRT7 OE+HG group (P<0.05). The results of flow cytometry showed that compared with the control group, the apoptosis rate of cells in the HG group was increased (P<0.05). Compared with the HG group, the apoptosis rate of cells in the siRNA SIRT7+HG group was increased significantly(P<0.05), while that in the SIRT7 OE+HG group was decreased (P<0.05). Compared with control group,the expressions of Nephrin, Wnt5a and β-catenin were inhibited in HG group (P<0.05). compared to HG group,siRNA-SIRT7 could down-regulate the expression levels of Nephrin, Wnt5a and β-catenin in siRNA-SIRT7 group (P<0.05), SIRT7 overexpression could up-regulate the expression levels of Nephrin, Wnt5a and β-catenin in SIRT7 OE+HG group (P<0.05). Conclusion: The findings suggest that high glucose environment is an important factor to inhibit the proliferation and induce apoptosis of mouse renal podocytes.Overexpression of SIRT7 can reverse the effects by activating Wnt/β-catenin signaling pathway and up-regulating β-catenin expression.

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