益气补肾方联合雷公藤多苷对糖尿病肾病大鼠Nephrin和Podocin表达的调控作用

Effect of Yiqi Bushen Prescription Together with GTW on Exprssion of Nephrin and Podocin in Rats with Diabetic Nephropathy

目的：观察益气补肾中药方联合雷公藤多苷（GTW）对糖尿病肾病（DN）大鼠肾脏的保护作用及其作用机制。方法：清洁级雄性SD大鼠70只,随机分成6组,应用高脂饲料喂养加单侧肾切除术合链脲菌素（STZ）腹腔注射制备DN大鼠模型。第9周起,西药组以厄贝沙坦（15.75 mg·kg-1·d-1）、雷公藤组以GTW（3.15 mg·kg-1·d-1）、中药组以益气补肾汤（相当于生药21.42 g·kg-1·d-1）、联合组以雷公藤多苷和益气补肾汤灌胃,治疗周期8周。16周末采用ELISA法检测大鼠尿微量白蛋白;应用血糖测定仪检测大鼠血糖水平;全自动血液细胞分析仪测定血白细胞;全自动生化仪测定大鼠肝肾功能;逆转录聚合酶链反应（RT-PCR）及免疫组化技术检测肾组织足细胞标志物Nephrin和Podocin的表达;光镜及电子显微镜观察肾组织病理变化。结果：与正常组比较,造模大鼠体重开始增加明显（0~6周）,但注射STZ后开始明显消瘦;血糖均显著上升（P〈0.01）;尿微量蛋白/肌酐比值（ACR）的显著升高（P〈0.05）;血肌酐及血尿素氮明显升高（P〈0.05）;Nephrin、Podocin蛋白及其m RNA表达均降低（P〈0.05）。光镜及电镜结果提示DN大鼠足细胞数目减少、足突增宽融合,系膜增宽,基底膜增厚。与模型组比较,各治疗组大鼠血Scr、BUN和尿ACR均有不同程度,同时Nephrin、Podocin蛋白及其m RNA的表达也相应有所上调,此外,各组病理改变也同时有改善,尤以联合治疗组效果最佳。结论：益气补肾方联合雷公藤多苷能对DN大鼠早期肾损伤具有保护作用,其作用机制可能与上调足细胞Nephrin和Podocin的表达有关,同时中药复方还具有抑制雷公藤多苷不良反应的功效。

Objective： To investigate the protection and mechanism of the Yiqi Bushen Prescription together with multi-glycoside from Tripterygium wilfordii Hook f.（ GTW） on diabetic nephropathy（ DN） rats. Methods： To establish an experimental DN rat model through a high-fat diet,unilateral nephrectomy and injection of streptozocin（ STZ） through peritoneal cavity. Animals were respectively administered with ARB [15. 75 mg /（ kg. d） ],GTW [3. 15 mg /（ kg · d） ],Yiqi Bushen Decoction [contained 21.42 g /（ kg·d） dried medicinal herbs]and the mixture of GTW and Yiqi Bushen Decoction from 9th week to 16 th week. At the end of 16 th week,the level of microalbumin（ MA） in urine was determined by ELISA and the level of GLU in serum determined by glucometer and the functions of liver and kidney were measured with Automatic Analyzer. The number of WBC was measured with hematology analyzer. The expressions of Nephrin and Podocin in renal cortex were determined with methods of RT-PCR and immunohistochemical method. The pathological changes of kidney were detected with light microscope and electron microscope. Results： Compared with normal rats,the body weight of DN rats increased quickly from 0th to 6th week but decreased after the rejection of STZ. The level of serum GLU was also increased significantly（ P〈0.01）. The ratio of MA and creatinine（ ACR） in urine was increased significantly（ P〈0.05）. The levels of Scr and BUN in serum were increased significantly（ P〈0.05）. The protein and m RAN expressions of Nephrin and Podocin in renal cortex were decreased significantly（ P〈0.05）. The pathologic change showed the numberof podocyte decreased,foot processes broadening and integrating,mesangial area broadening and basement membrane thickening. After the treatment,the level of Scr,BUN and ACR were decreased,accompanied with the increased expressions of Nephrin and Podocin and pathologic changes improved,especially in union of Yiqi Bushen Decoction and GTW treantment groups. Conclusion： The combination of Yiqi Bushen Decoction and GTW has protection effects on early renal injury in rats with DN and the treatment mechanism might be associated with the up adjustment of the expressions of Nephrin and Podocin. At the same time,Yiqi Bushen Decoction has a antagonistic action to toxic and side effects of GTW.