肾病一号方调控足细胞ANGPTL3表达的机制研究

ANGPTL3 over－expression in podocytes is closely related to podocyte injury , ANGPTL3 over－expression led to podocyte skeleton disorders, increased motility, increased urinary protein, while knockdown ANGPTL3 expression in podocytes reduced Adriamycin－induced podocyte injury. Moreover, ANGPTL3 knockout in mice does not affect the development of the kidney and renal function, maintain normal podocyte ultrastructure, significantly reduced podocyte injury in adriamycin-induced nephropathy, decreased urinary protein. It is significantly important to actively discover the drugs of regulation of expression of ANGPTL3 in podocytes for preventing or minimizing podocyte injury, and Shenbing yihao prescription can effectively decrease proteinuria, reduce podocyte injury in clinical and adriamycin-induced nephropathy, but whether the protective role of shenbing yihao prescription in podocyte injury is through the regulation of ANGPTL3 expression in podocytes is needed further study. It is necessary to explore the relationship between the protective effect on podocyte injury and the change of ANGPTL3 expression in podocytes in vivo and in vitro, and further explore the mechanism of shenbing yihao prescription in regulating ANGPTL3 expression in podocytes.

ANGPTL3在足细胞中高表达与足细胞损伤密切相关，ANGPTL3高表达导致足细胞骨架紊乱，活动性增加，尿蛋白增多，敲低足细胞ANGPTL3表达减轻了阿霉素对足细胞的损伤；小鼠体内敲除ANGPTL3不影响肾脏的发育和肾功能，足细胞超微结构正常，显著减轻了阿霉素肾病模型小鼠的足细胞的损伤，有效减少尿蛋白；积极寻求调控ANGPTL3表达的药物对防止或减轻足细胞损伤具有重要意义。肾病一号方在临床和大鼠阿霉素肾病模型中能够有效减少尿蛋白，减轻阿霉素肾病大鼠的足细胞损伤，但肾病一号方在足细胞损伤过程中是否通过调控ANGPTL3表达而起到对足细胞的保护有待进一步研究，课题拟通过肾病一号方在阿霉素肾病模型及体外足细胞损伤研究中深入探索其对足细胞的保护作用与足细胞ANGPTL3表达变化的关系，研究肾病一号方在调控足细胞ANGPTL3表达中的分子机制。

背景 肾病一号方有效减少肾病患儿尿蛋白，而ANGPTL3高表达与尿蛋白增多关系密切，敲除ANGPTL3可显著减轻了阿霉素肾病模型小鼠足细胞损伤，减少尿蛋白，本研究旨在探讨肾病一号方在肾病模型中调控足细胞ANGPTL3表达的分子机制。通过尾静脉注射观察阿霉素后观察阿霉素肾病小鼠在肾病一号方治疗后的保护作用及黄芪甲苷和泽泻醇B对PAN诱导足细胞损伤的保护作用；结果：1.肾病一号方在阿霉素肾病模型中的保护作用：阿霉素肾病造模后第4周，野生型空白对照组尿pro/cre、野生型阿霉素肾病模型组尿pro/cre、野生型阿霉素肾病中药治疗组尿pro/cre、Angptl3-/-阿霉素肾病模型组尿pro/cre、Angptl3-/-阿霉素肾病中药治疗组尿pro/cre具有显著性统计学差异；光镜下各组小鼠肾小球结构均无明显病理变化，电镜下野生型阿霉素肾病模型组小鼠肾小球足细胞足突融合明显，中药干预组足细胞足突融合较轻；野生型阿霉素肾病模型组Angptl3水平升高（P<0.05）,中药干预组Angptl3水平较模型组降低（P<0.05）。2.黄芪甲苷及泽泻醇B对PAN诱导足细胞损伤的保护作用 PAN模型组足细胞细胞骨架呈短棒状无序排列，而PAN+AS-IV、PAN+23-AAB组足细胞细胞骨架排列趋于整齐，部分丝状结构相对完整；PAN损伤模型组足细胞Angptl3 mRNA表达显著增高，PAN+AS-IV实验组Angptl3 mRNA的表达无明显降低（p＞0.05）。PAN+23-AAB实验组Angptl3 mRNA的表达无明显降低（p＞0.05）；结论：肾病一号方能轻阿霉素肾病小鼠模型足细胞损伤，减少尿蛋白，体外试验显示肾病一号方中的黄芪甲苷及23-AAB能够减轻PAN足细胞损伤模型中足细胞骨架蛋白结构紊乱。