通络益肾方不同给药时期对DN大鼠足细胞损伤的影响  

Effect of Tongluoyishen Decoction Different Administration Period on Podocyte Injury of Diabetic Nephropathy Rats

作　　者：李小会[1] 雷根平[2] 李京涛[2] 王军[2] 樊珊杉 谷浩荣[1] 

机构地区：[1]陕西中医药大学基础医学院,咸阳712000 [2]陕西中医药大学附属医院,咸阳712000

出　　处：《中国中西医结合肾病杂志》2017年第4期296-300,I0004,I0005,共7页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：国家自然科学基金资助项目(No.81473671);陕西省中医药管理局科研基金资助项目(No.13-LC064);陕西中医药大学附属医院科研项目(No.2014-11)

摘　　要：目的:观察通络益肾方对糖尿病肾病(DN)大鼠足细胞超微结构及骨架蛋白表达的影响。方法:60只SD大鼠随机抽取10只为假手术组,余大鼠采用右肾切除+腹腔注射链脲佐菌素制备DN模型。成模后的大鼠随机分为DN模型组(模型组),缬沙坦治疗组(西药组),通络益肾方早期给药组(中药早期组)、6周给药组(中期给药组)、8周给药组(晚期给药组)。每组10只。西药组予缬沙坦7.2 mg/kg灌胃,中药组予通络益肾方水煎液13.6 g/kg灌胃。中药早期组在造模成功后即予中药灌胃,中药中期和晚期组分别在造模成功后第6周、第8周开始中药灌胃。假手术组及模型组予蒸馏水灌胃。每日1次,共6周。检测大鼠24 h尿蛋白定量(24 h PRO)、尿白蛋白、肾功能;透射电镜观察足细胞超微结构,免疫组化法检测α-actintin-4、synaptopodin、F-actin表达,RT-PCR检测Nephrin、CD_2AP、α-actinin-4、synaptopodin、F-actin mRNA的表达。结果:(1)与假手术组比较,模型组24 h PRO、尿白蛋白、血尿素氮、血肌酐均明显升高(P<0.01)。与模型组比较,西药组、中药早期组、中药中期组上述指标均明显下降(P<0.01);与中药早期组比较,中药中期和晚期组各检测指标均不同程度升高(P<0.01,P<0.05);(2)足细胞超微结构的变化:模型组足细胞结构不清、数目减少、密度降低,骨架纤维排列紊乱;西药组及中药各治疗组足细胞病变减轻,足细胞较为完整,足突排列相对整齐,细胞骨架纤维排列相对整齐,足突融合减轻;中药早期组优于中药中期组和中药晚期组;(3)免疫组化结果显示:与假手术组比较,模型组α-actintin-4、synaptopodin、Factin表达显著下降(P<0.01)。与模型组比较,西药组、中药早期组、中期组上述指标均明显升高(P<0.01,P<0.05),中药晚期组差异无统计学意义(P>0.05);(4)与假手术组比较,模型组Nephrin、CD2AP、α-actinin-4、synaptopodin、F-actin mRNA表达均明显下降(P<0.01)Objective：To study the effect of Tongluoyishen Prescription on podocyte ultrastructure and related proteins expression by using Unilateral nephrectomy and intraperitoneal injection of Streptozotocin （STZ） diabetic nephropathy （DN） rats model. Methods ：60 SD rats were randomly divided into shamoperation group （n = 10） , the other rats were treated with right nephrectomy and intraperitoneal injection of streptozotocin to prepare DN model. Successful model rats were randomly divided into DN group （ model group） , Valsartan treatment group （Western medicine group） , Tongluoyishen early administration group （early group） , 6 weeks treatment group （medium -term group） , 8 weeks treatment group （late group）. 10 rats in each group. Rats in the Western medicine group were administered with valsartan by gastrogavage, while the traditional chinese medicine group were treated with Tongluoyishen Decoction by gastrogavage. The early group were given Tongluoyishen Decoction after the success of modeling, the medium - term and late groups were given decoction at the 6th week and the 8th week after the model establishment. Sham - operation group and model group were given distilled water. Once a day, total of 6 weeks. 24 - hour urinary protein （24 h UPRO） , urinary albumin, serum creatinine and blood urea nitrogen of each group were detected, the ultrastructure of glomerular podocytes was observed by transmission electron microscopy, the expressions of α - actinin - 4, synaptopodin and F - actin were detected using immunohistochemistry. Nephtin, CD2AP, α- actinin -4, synaptopodin and F - actin mRNA in podocytes were detected by RT - PCR. Results： （ 1 ） Compared with the sham - operation group, the 24 h UPRO, urinary albumin, blood urea nitrogen and serum ereatinine in the model group were significantly increased （P 〈 0.01 ）. Compared with the model group, the above- mentioned indexes of the western medicine group, the early group and the mediumterm group were significantly dec

关 键 词：通络益肾方 糖尿病肾病 足细胞 超微结构 

分 类 号：R285.5[医药卫生—中药学]