促红细胞生成素抑制C3介导的肾小管上皮细胞间充质转分化-内科学(肾脏病)专业论文.docxVIP

中文摘要 目的：观察促红细胞生成素（EPO）对 C3a 介导的肾小管上皮细胞间充质转 分化的影响，以及促红细胞生成素对单侧输尿管梗阻大鼠肾脏纤维化的影响。 方法： 一 体外实验：将人近端肾小管上皮细胞（HK-2）分为 6 组：对照组、10U/ml EPO 组、3ng/mlTGF-β 组、3ng/ml TGF-β+10U/ml EPO 组、0.1μM C3a 组、 10U/ml EPO + 0.1μM C3a 组；分别用 RT-PCR、Western Blot 和细胞免疫荧光方 法检测 HK-2 细胞 α-SMA、E-cadherin、C3 mRNA 和蛋白的表达。 二 体内实验：建立肾单侧输尿管梗阻大鼠（UUO）模型，并把实验动物分 成假手术组、UUO 对照组、EPO 小剂量治疗组（100U/Kg EPO）和 EPO 大剂量 治疗组（1000U/Kg EPO），在 UUO 术后第 3 至 14 天隔天腹膜内注射给药，所有 大鼠在第 14 天给予甲烷麻醉后处死；观察肾组织病理变化；免疫组化检测各组 大鼠肾脏组织 αSMA、E-cadherin 和 C3 表达。 结果： 一 体外实验：C3a和TGFβ干预HK2细胞后，α-SMA mRNA和蛋白表达增 强，E-cadherin mRNA和蛋白表达减少，补体C3 mRNA和蛋白表达增强；而加入 EPO干预后，则可抑制C3a和TGFβ的上述作用。 二 体内实验：UUO大鼠肾经过EPO治疗后，肾组织C3和α-SMA表达明显 减弱，E-cadherin表达明显增强。 结论： 1.EPO可抑制C3a和TGFβ诱导的肾小管上皮细胞间充质转分化作用。 2. UUO大鼠肾间质补体C3表达增强，EPO可抑制肾间质补体C3表达，并抑 制肾小管上皮细胞EMT的产生，EPO对肾小管上皮细胞EMT的抑制作用可能是 通过介导C3的表达来实现的。 关键词：重组人促红细胞生成素；补体 C3；肾脏纤维化 The mechanism study of inhibtion against C3-mediated renal tubular epithelial cells mesenchymal transition by EPO Objective To identify the role of EPO in the C3a induced epithelial mesenchymal transition and the effect to renal fibrosis of unilateral uretreal obstructire rats. Methods In vitro experiment: The HK-2 cells were divided into 6 groups namely control group, 10U/ml EPO group, 3ng/mlTGF-βgroup, 3ng/ml TGF-β+10U/ml EPO group, 0.1μM C3a group and 10U/ml EPO + 0.1μM C3a group. The expressions of α-SMA, E-cadherin and C3 mRNA were investigated by applying RT-PCR, Western Blot and immunofluorescence, respectively. In Vivo experiment: The model of UUO rats was established, and all animal subjects were assigned to four groups: sham operation group, UUO control group, Low dose EPO（100U/Kg EPO） treatment group and high dose EPO （1000U/Kg EPO）treatment group. The subjects were given an injection of EPO from 3days after operation to the 14th day. All animal subjects were given methane to induce anesthesia and consequently executed. The pathological change of renal was observed, and immunohistochemistry was performed to measure the expression of SMA, E-c