化瘀通络中药对糖尿病肾病大鼠肾脏足细胞裂孔膜蛋白podocin、CD2AP的调节作用  

Regulatory Effect of Chinese Drugs for Stasis Removing and Collaterals Dredging on the Expressions of podocin and CD2AP in Podocyte Slit Diaphragm of Diabetic Nephropathy Rats

作　　者：方敬[1] 陈志强[1] 郭倩[1] 陈春宇[1] 王春宁[1] 谢田[1] 孙汝[1] 刘利飞 张肖[1] 

机构地区：[1]河北医科大学中西医结合教研室,石家庄050017

出　　处：《中国中西医结合杂志》2016年第7期835-841,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81373804)

摘　　要：目的探讨化瘀通络中药对糖尿病肾病大鼠肾脏足细胞裂孔膜蛋白podocin、CD2AP的调节作用。方法 40只大鼠采用高糖高脂饲料联合1%链脲佐菌素(streptozotocin,STZ)35 mg/kg腹腔注射复制糖尿病大鼠模型,将造模成功的36只大鼠按随机数字表法分为模型组、化瘀通络组和厄贝沙坦组,每组12只。另取10只正常大鼠作为正常组。化瘀通络组和厄贝沙坦组分别给予化瘀通络中药和厄贝沙坦灌胃,同时正常组和模型组等量蒸馏水灌胃。采用ELISA法检测不同时间点各组大鼠24 h尿蛋白定量;连续干预16周后分别检测各组大鼠体重(body weight,BW)、肾重(kidney weight,KW)、肾脏指数(kidney index,KI)、空腹血糖(fasting blood glucose,FBG)、血肌酐(serum creatinine,SCr)、血尿素氮(blood urea nitrogen,BUN)和尿酸(uric acid,UA);光镜和电镜下观察肾组织病理形态学变化;采用Western blot和Real-time PCR法检测肾脏足细胞裂孔膜蛋白podocin、CD2AP蛋白和mRNA表达。结果 (1)与正常组比较,模型组大鼠第4、8、12、16周24 h尿蛋白定量明显升高(P<0.01),造模后大鼠BW减轻,KI、FBG、SCr、BUN和UA水平均升高(P<0.01);与模型组比较,化瘀通络组和厄贝沙坦组第4、8、12、16周24 h尿蛋白定量明显降低,且化瘀通络组24 h尿蛋白定量较厄贝沙坦组减少更明显(P<0.05,P<0.01);造模后化瘀通络组和厄贝沙坦组BUN水平降低(P<0.05,P<0.01)。(2)肾脏病理显示:模型组肾脏结构紊乱,基底膜明显增厚,系膜重度增生,足突广泛融合;化瘀通络组和厄贝沙坦组肾脏病理改变均有不同程度减轻。(3)Western blot和Real-Time PCR显示:与正常组比较,模型组大鼠肾组织podocin、CD2AP蛋白和mRNA表达水平均降低(P<0.01);与模型组比较,化瘀通络组和厄贝沙坦组podocin、CD2AP蛋白和mRNA表达水平均升高(P<0.01);化瘀通络组podocin、CD2AP蛋白和CD2AP mRNA表达均明显高于厄贝沙坦组,差异有统计学意义(P<0.05)。结论Objective To observe the regulatory effect of Chinese drugs for stasis removing and collaterals dredging（CDSRCD） on the expressions of podocin and CD2 AP in podocyte slit diaphragm（SD） of diabetic nephropathy（DN） rats. Methods DN rat model was duplicated in 40 male Sprague-Dawley rats by feeding high fat high glucose diet combined with intraperitoneally injecting 1% streptozotocin（STZ, 35 mg/kg）. Totally 36 successfully modeled rats were divided into the model group, the CDSRCD group, and the irbesartan group according to random digit table, 12 in each group. Besides, another 10 normal rats were recruited as a normal group. Rats in the CDSRCDgroup and the irbesartan group were intragastrically fed with CDSRCD and irbesartan respectively. Rats in the normal group and the model group were fed with equal volume of distilled water at the same time. 24 h urine protein quantitation was detected using ELISA at various time points. Body weight（BW）, kidney weight（KW）, kidney index（KI）, fasting blood glucose（FBG）, serum creatinine（SCr）, blood urea nitrogen（BUN）, and uric acid（UA） in each group were detected after 16 weeks of intervention. The pathomorphological changes of renal tissue were observed under light microscope and electron microscope respectively. The protein and m RNA expressions of podocin and CD2 AP were detected by Western blot and Real-time PCR respectively. Results（1） Compared with the normal group, 24 h urine protein quantitation significantly increased at week 4, 8, 12, and 16, respectively（P 〈0. 01）. BW was decreased; KI and levels of FBG, SCr, BUN, and UA all increased after modeling（P 〈0. 01）. Compared with the model group, 24 h urine protein quantitation significantly decreased in the CDSRCD group and the irbesartan group at week 4, 8, 12, and16, respectively（P 〈0. 01）. Besides, it was more obviously reduced in the CDSRCD group than in the irbesartan group（P 〈0. 05, P 〈0. 01）. BUN level obviously decreased both in the CDSRCD gr

关 键 词：糖尿病肾病 化瘀通络中药 PODOCIN CD2AP 

分 类 号：R285.5[医药卫生—中药学]