C／EBPα对糖尿病肾病足细胞保护作用的研究  

Protective effects of C/EBPα on podocytes in diabetic nephropathy

作　　者：周芳芳 张沥文 刘剑 应霁 刘韵子 仲芳[1,2] 王伟铭 陈楠[1] Zhou Fangfang;Zhang Liwen;Liu Jian;Ying Ji;Liu Yunzi;Zhong Fang;Wang Weiming;Chen Nan(Institute of Nephrology, Shanghai Jiaotong University School of Medicine, Department of Nephrology, RuiJin Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai 200025, Chin)

机构地区：[1]上海交通大学医学院肾脏病研究所上海交通大学医学院附属瑞金医院肾脏科,200025 [2]美国纽约州西奈山伊坎医学院肾脏科

出　　处：《中华肾脏病杂志》2018年第5期355-360,共6页Chinese Journal of Nephrology

基　　金：国家自然科学基金（81270782、30771000）;国家重点基础研究发展计划（“973”计划）（2012CB517701、2012CB517604）;上海市科研计划项目（15140902800）

摘　　要：目的构建足细胞C／EBPα条件性敲除小鼠，进一步构建糖尿病肾病模型，研究C／EBPα敲除后对糖尿病肾病足细胞的影响及机制。方法将C／EBPαlwp/loxp与podocin—cre小鼠进行杂交，得到F1代杂合子后继续繁殖直至得到纯合子小鼠（C／EBPαf/f）。分别通过常规喂养或45％高脂饮食喂养25周，并用低剂量链脲佐菌素（STZ，100mg／kg）处理小鼠后构建糖尿病肾病模型，检测小鼠血尿生化指标以及观察各组肾脏组织形态学改变，并检测。肾组织纤维化、氧化应激调控因子和线粒体功能相关蛋白的表达。结果成功构建足细胞C／EBPα特异性敲除小鼠，并在此基础上成功构建2型糖尿病肾病模型。8月龄的C／EBPaf/f小鼠。肾脏无明显组织形态学异常，但糖尿病。肾病的C／EBPαf/f小鼠具有明显的肾脏损伤、炎症和氧化应激反应。与野生型糖尿病肾病小鼠相比，糖尿病肾病C／EBPαf/f小鼠Nephrin和E—cadherin表达极度降低（均p〈0．01），Fibronectin显著升高（P〈0．01），Nrf2显著升高（P〈0．05），Keapl降低（P〈0．05），且磷酸化AMPK水平降低（P〈0．05），线粒体功能相关基因Pgc-1α也显著降低（P〈0．05）。结论足细胞C／EBPα基因敲除通过促进纤维化并抑制Pgc—1α介导的线粒体抗氧化功能加重糖尿病肾病。Objective To explore the effects of C/EBPα knockout in podocyte on diabetic nephropathy and its mechanisms. Methods C/EBPloxpl/oxp mice were crossed with podocin-cre mice to obtain F1 hybrids and then propagated until homozygous mice （C/EBPαf/f） were obtained. Diabetic nephropathy （DN） models were established by low-dose streptozotocin （STZ, 100 mg/kg） administration after 25 weeks of normal diet or 45% high-fat diet treatment, and biochemical indicators of blood and urea were measured. The morphological characteristics and the proteins regulating oxidative stress and mitochondrial function were detected. Results The type 2 DN models were successfully constructed based on transgenic mice. The kidneys of 8- month- old C/EBPαf/f mice did not show obvious morphological changes, but after constructing DN models, they showed obvious renal impairment, inflammation and oxidative stress. Compared with wild-type DN mice, the protein levels of nephrin and E-eadherin in DN C/EBPαf/f mice with DN were significantly decreased （P 〈 0.01）; fibronectin and Nrf2 protein levels were all increased （all P 〈 0.05）. Keapl, phospho-AMPK and mitochondrial function- related genes Pge- 1α protein levels were all decreased （all P 〈 0.05）. Conclusion Podocyte C/EBPα knockout exacerbates diabetic nephropathy by promoting fibrosis and inhibiting Pgc- 1α- mediated mitochondrial antioxidant function.

关 键 词：糖尿病肾病 足细胞 小鼠 基因敲除 CCAAT增强子结合蛋白 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]