肾痿复方对糖尿病肾病大鼠肾组织MMP-9/TIMP-1表达的影响  

Effect of Shenwei Compound on the expression of MMP-9/TIMP-1 in the renal tissue of rats with diabetic nephropathy

作　　者：钟霞 张宇韦 谭睿陟 樊均明[2] 王丽 ZHONG Xia;ZHANF Yuwei;TAN Ruizhi;FAN Junming;WANG Li(Research Center of Integrated Chinese and Western Medicine, the Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China;Chengdu Medical College)

机构地区：[1]西南医科大学附属中医医院中西医结合研究中心,四川泸州646000 [2]成都医学院

出　　处：《西南医科大学学报》2019年第5期413-418,共6页Journal of Southwest Medical University

基　　金：四川省教育厅科研创新团队(项目编号:17TD0046);泸州市-西南医科大学联合项目-泸州市青年创新团队(2016LZXNYD-T05)

摘　　要：目的:通过对糖尿病肾病大鼠肾功能及肾组织中α平滑肌肌动蛋白(α-SMA)、转化生长因子-β1(TGF-β1)、基质金属蛋白酶-9(MMP-9)及基质金属蛋白酶抑制物-1(TIMP-1)水平的检测,探讨肾痿复方在糖尿病肾病的治疗作用。方法:40只雄性SD大鼠随机为正常对照组、糖尿病肾病组、肾痿复方低、高剂量组、每组各10只。糖尿病肾病组大鼠经高脂高糖饲料喂养2月后,腹腔注射链脲佐菌素(35mg/kg)诱导建立糖尿病肾病模型,72h后测定空腹血糖≥16.65mmol/L者视为造模成功。正常组、糖尿病肾病组大鼠予以生理盐水灌胃,低剂量治疗组、高剂量治疗组大鼠分别予以肾痿复方灌胃。给药1周后,检测24h尿蛋白、血清肌酐、尿素氮,HE和PAS染色检测肾脏病理组织改变,免疫印记法(WesternBlot)检测α-SMA、TGF-、MMP-9及TIMP-1蛋白表达。结果:①肾功:与糖尿病肾病组比较,低剂量组、高剂量组大鼠血肌酐、尿素氮水平降低,差异有统计学意义(P<0.05)。②肾脏形态学:HE和PAS染色显示:正常组大鼠肾小球形态结构正常,系膜细胞及基质无增生;糖尿病肾病组大鼠肾组织可见炎性细胞浸润,肾小球系膜细胞增生,基底膜增厚,肾小管上皮细胞空泡变性,管腔扩张。低、高剂量组大鼠可见肾小球、肾小管、肾间质病理损伤程度减轻,其中高剂量治疗组大鼠肾脏损害明显改善。③蛋白水平:与模型组比较,肾痿复方低、高剂量组大鼠随着药物浓度的增加肾组织中α-SMA、TGF-β1、TIMP-1表达逐渐降低(P<0.05、P<0.01或P<0.001)、而MMP-9随着药物浓度的增加表达逐渐升高(P<0.05)。结论:肾痿复方可能通过提高MMP-9,降低α-SMA、TGF-β1、TIMP-1的表达,从而减轻糖尿病肾病引起的肾脏病理损害。Objective: To explore the therapeutic effect of Shenwei Compound on diabetic nephropathy by testing the renal function of rats with diabetic nephropathy(DN) and determining the levels of alpha-smooth muscle actin(α-SMA), transforming growth factor-β1(TGF-β1), matrix metalloproteinase-9(MMP-9), and tissue inhibitor of metalloproteinase-1(TIMP-1) in the renal tissue of the rats. Methods: Forty male Sprague-Dawley rats were randomly divided into normal control group, DN group, and low-and high-dose groups of Shenwei Compound, with 10 rats in each group. After two months of high-fat and high-sugar diet feeding, a rat model of DN was established in the DN group by intraperitoneal injection of streptozotocin(35 mg/kg). The modelling was considered successful if fasting blood glucose tested after 72 hours was ≥16.65 mmol/L. Rats in the normal group and the DN group were administered intragastrically with normal saline, and rats in the low-and high-dose groups were administered intragastrically with Shenwei Compound. After 1 week of administration, the rats were tested for the levels of 24-h urinary protein, serum creatinine, and urea nitrogen;meanwhile, pathological changes in the renal tissue were evaluated by HE and PAS staining;Western blot was used to determine the protein expression of α-SMA, TGF-β1, MMP-9, and TIMP-1. Results: Renal function: Compared with the DN group, the low-and high-dose groups had significantly reduced levels of serum creatinine and urea nitrogen(P < 0.05). Renal morphology: HE and PAS staining showed that the morphological structure of the glomeruli of the rats in the control group was normal, without any proliferation of mesangial cells or matrix;renal tissue staining of the rats in the DN group showed inflammatory cell infiltration,proliferation of glomerular mesangial cells, thickening of the basement membrane, and vacuolar degeneration of renal tubular epithelial cells and dilatation of the lumen. A reduced degree of pathological damage was observed in the glomerulus, renal tubule

关 键 词：肾痿复方 糖尿病肾病 疾病模型 Α平滑肌肌动蛋白 转化生长因子β1 基质金属蛋白酶-9 基质金属蛋白酶抑制物-1 

分 类 号：R587.1[医药卫生—内分泌] R332[医药卫生—内科学]