中介素对缺氧肾小球内皮细胞管腔样结构形成的影响

Effect of intermedin on tube-like structure formation of glomerular endothelial cells under hypoxic condition

目的研究中介素(IMD)对缺氧条件下大鼠肾小球内皮细胞(r RGECs)管腔样结构形成的影响和可能机制。方法在体外培养传代rRGECs后分为正常对照组、缺氧组、IMD组,后2组放置5%O2缺氧环境中培养6 h,对照组在正常氧浓度中培养,四甲基偶氮唑盐(MTT)检测细胞存活;小管形成实验检测管腔样结构形成;免疫细胞化学检测血管内皮生长因子(VEGF)、血管内皮钙黏着蛋白(VE-cadherin)蛋白表达;实时荧光定量聚合酶链反应(Q-PCR)检测VEGF、VE-cadherin mRNA表达水平。结果与正常对照组相比,缺氧组细胞增殖率,管腔样结构的管腔长度、成环数和分支数以及VE-cadherin mRNA和蛋白表达下降(P<0.05),VEGF mRNA和蛋白表达升高(P<0.05);与缺氧组相比,IMD组细胞存活率,管腔样结构的管腔长度、成环数和分支数以及VEGF、VE-cadherin mRNA和蛋白表达升高(P<0.05)。结论在缺氧条件下IMD可促进r RGECs增殖和管腔样结构形成,减轻r RGECs损伤,并可能与VEGF、VE-cadherin有关。

Objective To investigate the effects and potential mechanism of intermedin（IMD） on tube-like structure formation of rat renal glomerular endothelial cells（rRGECs） under hypoxic condition. Methods After cultured in vitro, the subcultured rRGECs were randomly divided into three groups, the normal control group, hypoxia group and IMD group. The latter two groups were cultured for 6 h in the hypoxic environment of 5% O2. The control group was cultured in the normoxic environment. The cell viability was examinated by MTT assay. The tube-like structure formation was determined by tube formation assay. The protein expression of vascular endothelial growth factor（VEGF） and vascular endothelial-cadherin（VE-cadherin） were measured by immunocytochemistry. The mRNA expression levels of VEGF and VE-cadherin were determined by real time fluorescent quantitative PCR（qPCR）. Results Compared with the normal control group, the cell proliferation rate, length of the tube-like lumen, number of loops and branches, and the mRNA and protein expression of VE-cadherin in the hypoxia group were all decreased（P〈0.05）,whereas the mRNA and protein expression of VEGF were increased（P〈0.05）. Compared with the hypoxia group, the cell viability, length of the tube-like lumen, number of loops and branches, and the mRNA and protein expression of VEGF anc VE-cadherin were increased in the IMD group（P〈0.05）. Conclusion IMD can promote the proliferation of r RGECs and the formation of tube-like structure under hypoxic condition, and reduce the injury of r RGECs, which may be related to VEGF and VE-cadherin.