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[摘要]
目的 观察复肾汤(FST)对糖尿病肾病(DN)大鼠足细胞nephrin和podocin表达的影响,并探讨其可能的作用机制。方法 正常对照组为雄性Wistar大鼠8只,常规饲养。雄性Wistar大鼠24只,正常饲养1周后大鼠ip链脲佐菌素(STZ)和弗氏完全佐剂(CFA)建立糖尿病模型。造模成功后分为模型组、FST大剂量组(L1)、FST小剂量组(L2)。L1、L2组分别给予复肾汤32、8 g/kg;模型组给予生理盐水。每天ig给药1次,给药体积1 mL/100 g,连续8周,定期测血糖。实验结束后,收集24 h尿液,采集血清,检测胰岛素(INS)、肌酐(SCr)、尿素氮(BUN)、超氧化物歧化酶(SOD)、丙二醛(MDA)、尿蛋白(UP)等生化指标;轻取肾脏,光镜下观察肾脏组织病理变化;免疫荧光法检测肾组织nephrin和podocin的表达,RT-qPCR法检测肾组织nephrin、podocin的基因表达。结果 与模型组比较,L1、L2组的INS、SCr、BUN、MDA、UP明显降低(P<0.05、0.01),SOD明显升高(P<0.05),病理学评分明显降低(P<0.05),nephrin、podocin的免疫荧光表达明显上调,RT-qPCR基因表达明显上调。结论 复肾汤可以有效改善DN大鼠足细胞nephrin和podocin的表达,对DN的发展具有一定的延缓作用。
[Key word]
[Abstract]
Objective To observe the effects of Fushentang on nephrin and podocin in podocytes of diabetic nephropathy rats and discuss the possible mechanism. Methods The normal control group was eight male Wistar rats with conventional feeding. A total of 24 male Wistar rats were treated with intraperitoneal injection of STZ + CFA to establish diabetic model after normal feeding for 1 weeks. After successful modeling, the rats were divided into model group, FST supernatant group (L1), FST sediment group (L2). L1, L2 group were treated with FST supernatant and sediment. The model group was given normal saline. Rats were administered ig for once daily with a volume of 1 mL/100 g, and blood glucose was measured regularly for 8 weeks. After the experiment, 24 h urine, the serum were collected, and the biochemical parameters, like INS, SCr, BUN, SOD, MDA, UP were detected. The kidney was removed and the pathological changes were observed under light microscope, the expressions of nephrin and podocin in renal tissues were detected by immunofluorescence, and the gene expressions of nephrin and podocin in renal tissues were detected by RTqPCR.Results Compared with the model group, the INS, SCr, BUN, MDA, UP of L1 and L2 groups were significantly decreased (P<0.05, 0.01), and SOD was increased significantly (P<0.05), the pathology score decreased significantly (P<0.05). Immunofluorescence expressions of nephrin and podocin were significantly up-regulated, and gene expressions of RT-qPCR were significantly up-regulated. Conclusion Fushentang can significantly improve the expression of nephrin and podocin in podocytes of DN rats, and delay the development of diabetic nephropathy.
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[基金项目]
江苏省自然科学基金青年基金项目(BK20180827)