通络方剂对糖尿病大鼠肾脏p38 MAPK-FN通路的影响

目的研究通络方剂(TLR)对链脲佐菌素(STZ)诱导的糖尿病大鼠肾脏p38 MAPK-纤维连接蛋白(FN)信号转导通路的影响,探讨TLR对肾脏保护作用的可能机制。方法将SD大鼠随机分成3组(n=7):正常对照组(CON组)、糖尿病未干预组(DM组)和糖尿病TLR干预组(DM+TLR组),DM组和DM+TLR组予以STZ(65 mg/kg)腹腔注射诱导糖尿病模型。制模成功后,DM+TLR组大鼠予以1 g/(kg.d)TLR灌胃,CON组和DM组用等剂量的双蒸水灌胃,持续12周。用自动化生化分析仪检测生化指标,用考马斯亮蓝法定量测定24 h尿蛋白,肾皮质进行PAS染色,用蛋白印迹分析法检测t-p38MAPK、p-p38 MAPK、t-CREB、p-CREB、FN的表达。结果与CON组相比,DM组大鼠的左肾质量/体质量、血糖、尿素氮、血肌酐、24 h尿蛋白、尿蛋白/肌酐增加(P<0.05),体质量减少(P<0.05),p-p38 MAPK、p-CREB、FN表达水平分别增加2.37倍、2.21倍、2.03倍(P<0.05);与DM组相比,DM+TLR组左肾质量/体质量、血肌酐、24 h尿蛋白、尿蛋白/肌酐降低(P<0.05),p-p38 MAPK、p-CREB、FN表达水平降低(P<0.05)。结论 TLR可能通过抑制p38 MAPK-FN通路对糖尿病肾病大鼠的肾脏产生保护作用。

Effects of Tongluo recipe on p38 mitogen-activated protein kinase-fibronectin pathway in the kidney of diabetic rats

ObjectiveTo study the effects of Tongluo recipe on p38 mitogen-activated protein kinase(p38 MAPK)-fibronectin(FN) signal pathway in the kidney of streptozotocin-induced diabetic rats, so as to explore the possible mechanism by which Tongluo recipe protects the kidney. MethodsMale Sprague-Dawley(SD) rats were randomly divided into 3 groups(n=7): control group (CON group), model group(DM group)and Tongluo recipe group(DM+TLR group). Rats in the DM group and DM+TLR groups were administered with streptozotocin (65 mg/kg) by intraperitoneal injection to induce diabetic model. Rats in DM+TLR group were also given 1 g/(kg·d) of Tongluo recipe orally after successfully establishment of diabetic model, and rats in DM and CON groups were only given equivalent volume of ddH2O. The interference lasted for 12 weeks. Biochemical indices were obtained by automatic analyzer. Coomassie brilliant blue assay was used to measure the 24-hour urine protein. Kidney cortex was subjected to PAS staining. Western blotting analysis was used to detect the protein expression of total p38 MAPK (t-p38 MAPK), phosphorylated p38 MAPK (p-p38 MAPK), total cAMP response element-binding protein (t-CREB), phosphorylated CREB (p-CREB) and FN. ResultsCompared with CON group, the average left kidney mass/body mass (LKM/BM), blood glucose(BG), blood urea nitrogen(BUN), serum creatinine(Scr), 24-hour urine proteins(24 h UP) and urine protein/creatinine(UP/Ucr) were significantly increased in the DM group (P<0.05); the average BM was significantly decreased(P<0.05); and the expression of p-p38 MAPK, p-CREB, FN was increased by 2.37, 2.21, and 2.03 folds, respectively (P<0.05). Compared with DM group, the average LKM/BM, Scr, 24 h UP and UP/Ucr were significantly decreased (P<0.05) in DM+TLR group, and the expression of p-p38 MAPK, p-CREB, FN was significantly decreased(P<0.05). ConclusionTongluo recipe can protect the kidney in diabetic nephropathy rats, probably through inhibition of the p38 MAPK-FN pathway .