PPARγ基因过表达/沉默对缺氧诱导的肾小管上皮细胞坏死性凋亡标志物表达调控作用  

Regulatory effect of overexpression/silencing of PPARγ gene on expression of necrotic apoptosis markers in renal tubular epithelial cells induced by hypoxia

作　　者：蹇淑娟 涂立 邹家森 朱仕群 覃远汉[1] JIAN Shujuan;TU Li;ZOU Jiaseng;ZHU Shiqun;QIN Yuanhan(The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院,广西南宁530021

出　　处：《山东医药》2021年第6期22-25,共4页Shandong Medical Journal

基　　金：广西自然科学基金资助项目(广西科学基金);广西自然科学基金项目(2017GXNSFDA198008)。

摘　　要：目的观察过氧化物酶体增殖物激活受体γ(PPARγ)基因过表达/沉默对缺氧诱导肾小管上皮细胞(RTEC)坏死性凋亡标志物表达的调控作用。方法将RTEC细胞随机分为对照组、缺氧损伤组、过表达组和沉默组,过表达组细胞转染过表达PPAYγ基因的外源性LV-Pparg病毒,沉默组细胞转染沉默PPAYγ基因的内源性LVPPARg-RNAi病毒。待细胞再次生长融合至70%左右时,正常对照组不做任何处理,放置于培养箱中培养;缺氧损伤组、过表达组以及沉默组放置于含95%N_(2)和5%CO_(2)混合气体缺氧小室中,密封缺氧培养48 h。采用实时荧光定量PCR法检测各组细胞中PPARγ、受体交互作用蛋白1(RIP1)、RIP3、转化生长因子-β(TGF-β)mRNA,采用Westernblotting法检测各组细胞中PPARγ、RIP1、RIP3、TGF-β蛋白。结果与对照组相比,缺氧损伤组、过表达组、沉默组细胞中PPARγ的mRNA及蛋白相对表达量均显著降低(P均<0.05),RIP1、RIP3、TGF-β的mRNA及蛋白相对表达量均显著升高(P均<0.05)。与缺氧损伤组相比,过表达组细胞中PPARγ的mRNA及蛋白相对表达量均显著升高(P均<0.05),RIP1、RIP3、TGF-β的mRNA及蛋白相对表达量均显著降低(P均<0.05);沉默组细胞中PPARγ的mRNA及蛋白相对表达量均显著降低(P均<0.05),RIP1、RIP3、TGF-β的mRNA及蛋白相对表达量均显著升高(P均<0.05)。结论在缺氧诱导的RTEC坏死性凋亡中,PPARγ基因表达降低;过表达PPARγ基因对缺氧诱导的RTEC坏死性凋亡有抑制作用,而沉默PPARγ基因则有促进作用。Objective gene overexpression/silencing on the expression of necrotic apoptosis markers in the renal tubular epithelial cells(RTECs)induced by hypoxia.Methods expression group,and silence group,and the cells in the overexpression group were transfected with exogenous LV-PPARg virus overexpressing PPARγgene;and the cells in the silence group were transfected with endogenous LV-PPARg-RNAi virus silencing PPARγgene.When the cells grew and converged to about 70%,the cells in the normal control group were cultured in the incubator without any treatment;the cells in the hypoxia injury group,overexpression group,and silence group were placed in an anoxic chamber containing mixed gases of 95%N2 and 5%CO2,and then were cultured in sealed and hypoxic environment for 48 h.Real-time fluorescence quantitative PCR was used to detect PPARγ,receptor-interacting protein 1(RIP1),RIP3,transforming growth factor-β(TGF-β)mRNA,and Western blotting was used to detect PPARγ,RIP1,RIP3,and TGF-βprotein in each group.Results mRNA and protein expression levels of PPARγin the hypoxia injury group,overexpression group,and silence group decreased significantly(all P<0.05),while the relative mRNA and protein expression levels of RIP1,RIP3,and TGF-βincreased significantly(all P<0.05);compared with the hypoxia injury group,the relative mRNA and protein expression levels of PPARγin the overexpression group significantly increased(all P<0.05),while the relative mRNA and protein expression levels of RIP1,RIP3,and TGF-βdecreased in the overexpression group(all P<0.05).In the silence group,the relative mRNA and protein expression levels of PPARγdecreased significantly(both P<0.05),while the relative mRNA and protein expression levels of RIP1,RIP3,and TGF-βincreased significantly(all P<0.05).Conclusion crotic apoptosis of RTEC induced by hypoxia,the expression of PPARγgene decreases;overexpression of PPARγgene can inhibit the necrotic apoptosis of RTEC induced by hypoxia,while silencing PPARγgene can promote it.

关 键 词：过氧化物酶体增殖物激活受体Γ 基因调控 肾小管上皮细胞 坏死性凋亡 缺氧损伤 

分 类 号：R692.6[医药卫生—泌尿科学]