| 摘要: |
| 目的 研究通络方剂对链脲佐菌素(STZ)诱导的1型糖尿病大鼠肾脏组织ERK1/2信号转导通路的影响及对肾脏的保护机制。方法 健康SD大鼠随机分为正常组(10只)和造模组。造模组给予单次腹腔注射STZ 60 mg/kg,造模成功后按体质量随机分为5组：糖尿病非治疗组，通络方剂低剂量组[0.5 g/(kg·d)]、中剂量组[1.0 g/(kg·d)]、高剂量组[2.0 g/(kg·d)]和氯沙坦组[30 mg/(kg·d)],每组10只。适应性饲养1周后,按分组分别给予不同处理,持续给药12周后处死,称体质量,取血检测血糖、血脂、血肌酐、尿素氮,留24 h尿检测尿肌酐、尿蛋白、尿微量白蛋白,ELISA法检测血清及肾脏组织中血管紧张素Ⅱ(Ang Ⅱ)的含量,取左肾称质量,H-E、PAS染色观察肾脏病理改变,免疫组织化学法检测p-ERK1/2蛋白的表达,蛋白质印迹法检测VEGF蛋白的表达。结果 与糖尿病非治疗组比较,糖尿病大鼠系膜细胞增殖受到抑制,蛋白尿明显降低,体质量增加,肾肥大指数降低,肌酐清除率、血尿素氮降低,肾功能改善,血清及肾组织Ang Ⅱ含量降低,VEGF及p-ERK1/2蛋白的表达降低(P＜0.05)。中剂量通络方剂与氯沙坦在改善糖尿病大鼠肾功能及抑制Ang Ⅱ、VEGF、p-ERK1/2表达方面差异无统计学意义。结论通络方剂对糖尿病大鼠肾脏的保护作用可能与抑制Ang Ⅱ、VEGF表达,阻断ERK1/2信号转导通路有关。 |
| 关键词: 糖尿病肾病 通络方剂 细胞外信号调节蛋白激酶 血管紧张素Ⅱ 血管内皮生长因子 |
| DOI：10.3724/SP.J.1008.2011.0949 |
| 投稿时间：2011-01-05修订日期：2011-06-27 |
| 基金项目:国家重点基础研究发展计划(“973”计划, 2005CB523304). |
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| Effects of Tongluo recipe on renal expression of Ang Ⅱ, VEGF, and ERK1/2 in rat models of type 1 diabetes mellitus |
| TANG Wen-jia,ZOU Jun-jie,SHI Yong-quan,XING Yu-wei,LIU Zhi-min* |
(Department of Endocrinology, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
*Corresponding author.) |
| Abstract: |
| ObjectiveTo observe the effects of Tongluo recipe on extracellular signal-regulated protein kinase (ERK1/2) pathway in the kidneys of streptozotocin-induced rat models of type 1 diabetes mellitus, and to explore the renoprotective mechanism of Tongluo recipe. MethodsAdult male Sprague-Dawley rats were randomly divided into 2 groups: control group(A, n=10) and experimental group. Rats in the experimental group were induced by intraperitoneal injection of streptozotocin (60 mg/kg body mass), and were randomly subdivided into group B (diabetic control, n=10), group C (diabetic rats treated with low dose of Tongluo recipe,0.5 g/\[kg·d\], n=10), group D (diabetic rats treated with medium dose of Tongluo recipe,1.0 g/\[kg·d\], n=10), group E (diabetic rats treated with high dose of Tongluo recipe,2.0 g/\[kg·d\], n=10) and group F (diabetic rats treated with losartan, 30 mg/\[kg·d\],n=10). At the end of the 12th week rats were sacrificed, and the serum glucose, triacylglycerol(TG), total cholesterol(Chol), serum creatinine(Scr), urea nitrogen(BUN) in blood, 24-hour urine protein(Upro24), microalbuminuria (MAU), and creatinine in urine(UCR) were examined. The kidney mass/body mass (KM/BM) and creatinine clearance rate (Ccr) were calculated. The left kidneys were collected, weighed and subjected to H-E staining and PAS staining to observe the pathological changes; serum and renal tissue angiotensin Ⅱ(Ang Ⅱ) levels were determined by ELISA; the expression of phosphorylated ERK1/2(p-ERK1/2) was detected by immunohistochemical staining and vascular endothelial growth factor(VEGF) protein expression was determined by Western blotting analysis. ResultsCompared with group B, group D had a slighter mesangial proliferation under light microscope. Besides, rats in group D had significantly decreased Ccr, BUN, MAU, and Upro24 in the serum, and decreased Ang Ⅱ in the serum and renal tissue, and decreased KM/BM, VEGF and p-ERK1/2 levels in renal tissue compared with rats in group B(P<0.05). The medium dose of Tongluo recipe and losartan showed similar effects in improving the renal function and inhibiting Ang Ⅱ, VEGF, and p-ERK1/2 expression of diabetic rats. ConclusionThe renoprotective effect of Tongluo recipe may be related to inhibition of ERK1/2 cascade pathway by decreasing the expression of Ang Ⅱ and VEGF. |
| Key words: diabetic nephropathies Tongluo recipe extracellular signal-regulated protein kinase angiotensin Ⅱ vascular endothelial growth factor |
