藏药灰兜巴对糖尿病肾病大鼠足细胞损伤的改善作用

Ameliorative effects of Tibetan Huidouba on podocyte injury in diabetic nephropathy rats

目的探讨藏药灰兜巴对糖尿病肾病大鼠足细胞损伤的改善作用。方法采取高脂高糖饮食喂养和腹腔注射65 mg/kg链脲佐菌素法诱导糖尿病肾病大鼠模型。大鼠随机分为对照组、模型组、二甲双胍组、灰兜巴组,每组8只,灌胃给药6周。分别于药物干预前后进行口服葡萄糖耐量实验(oral glucose tolerance test,OGTT),收集24小时尿液并检测mAlb与Ucr比值;PASM观察肾组织病理学改变情况;Western blot法和免疫荧光法检测足细胞标志蛋白Nephrin表达;透射电子显微镜观察足细胞和肾小球基底膜损伤。结果与对照组比较,模型组大鼠时间-血糖曲线下面积及mAlb/Ucr升高(P<0.01),足细胞标志蛋白Nephrin表达减少(P<0.01),透射电镜下可见肾小球基底膜明显增厚及广泛足突融合。与模型组比较,灰兜巴可改善糖尿病肾病大鼠空腹葡萄糖耐量受损,降低mAlb/Ucr(P<0.01),上调足细胞标志蛋白Nephrin表达(P<0.01),减少足细胞丢失,改善足细胞和肾小球基底膜超微结构改变情况。结论益气养阴藏药灰兜巴可减轻糖尿病肾病大鼠足细胞损伤,降低蛋白尿,从而发挥肾脏保护作用。

AIM To explore ameliorative effects of Huidouba(HDB),a Tibetan medical agent,on podocyte injury in diabetic nephropathy(DN)rats.METHODS Rats were induced into DN models by high fat diet(HFD)feeding and intraperitoneal injection of 65 mg/kg streptozocin(STZ).Rats were randomly divided into control group,model group,metformin group,and HDB group,8 rats in each group,for 6-week corresponding gavage.Oral glucose tolerance test(OGTT)was conducted before and after drug intervention,and 24-hour urine was collected to detect mAlb and Ucr ratio.Renal histopathology analysis was performed by PASM staining.Nephrin,the biomarker of podocyte,was examined by Western blot and immunofluorescence assay of renal tissue.Podocyte injury and ultrastructural changes of glomerular basement membrane(GBM)were detected by transmission electronic microscope(TEM).RESULTS Compared with the control group,the model group shared more area under glucose-time curve(AUC)and higher mAlb/Ucr level(P<0.01),but decreased Nephrin expression(P<0.01),obviously increased GBM thickness,and more extensive fusion of foot processes under TEM.Compared with the model group,HDB group shared improvement in the impaired fasting glucose tolerance and mAlb/Ucr reduction in DN rats(P<0.01),up-regulation of renal Nephrin expression,and therefore resultant improvement in ultrastructure damage of podocyte and GBM.CONCLUSION HDB,a Tibetan tonic for Benefiting Qi and Nourishing Yin,can improve podocyte damage and proteinuria,and thus protect kidney from damage in DN rats.