芡实对糖尿病肾病大鼠肾组织SOCS-3、IGF-1表达的影响

发布时间：2018-05-16 17:35

本文选题：糖尿病肾病 + 芡实　；参考：《山西医科大学》2014年硕士论文

【摘要】：目的：观察芡实对糖尿病肾病(diabetic nephropathy, DN)大鼠肾组织中SOCS-3及IGF-1表达的影响，进一步探讨芡实延缓糖尿病肾病进展的可能机制。方法：健康清洁级雄性SD大鼠60只，适应环境1周后，用随机数字表法抽取10只为正常对照组(NC组)，其余50只大鼠采用单剂量腹腔注射链脲佐菌素(STZ45mg/Kg)以建立糖尿病大鼠模型，随机分为糖尿病肾病组(DN组)、糖尿病肾病+小剂量芡实治疗组(EL组1.5g/kg/d)、糖尿病肾病+中剂量芡实治疗组(EM组3.0g/kg/d)、糖尿病肾病+大剂量芡实治疗组(EH组6.0g/kg/d)及糖尿病肾病+氯沙坦钾治疗组(LP组30mg/kg/d)，正常组腹腔注射相同体积不含STZ的柠檬酸钠缓冲液。造模组大鼠于72h后断尾测血糖，若血糖值16.7mmol/L，认为糖尿病模型成立；3周后测24h尿蛋白定量30mg/d，认为DN模型成功。各实验组给予相应剂量药物灌胃治疗，模型组及对照组给予等体积的蒸馏水灌胃，治疗12周。于治疗12周末记录各组大鼠的体重，留取24小时尿液标本及空腹血标本，分别用来检测24小时尿蛋白定量、血清尿素氮(BUN)及血肌酐(Scr)，然后经腹腔注射水合氯醛麻醉大鼠，逐层打开腹腔，经腹主动脉取血，冲洗肾脏至灰黄色，留取肾组织标本采用HE染色、Masson染色、PAS染色及电镜观察肾组织的病理变化，采用免疫组织化学法及Western blot技术检测各组大鼠肾组织中SOCS-3与IGF-1的表达及分布情况。结果：(1).生化参数的比较：治疗12周末，NC组大鼠血糖、血尿素氮、血肌酐、24h尿蛋白定量均在正常范围内波动；与NC组相比，，DN组大鼠血尿素氮、血肌酐、24h尿蛋白定量明显增加(P0.05)；与DN组对比，EM组、EH组及LP组大鼠血尿素氮、血肌酐、24h尿蛋白定量显著降低(P0.05)；(2).肾脏病理改变：12周末，在光镜、电镜下可见，NC组大鼠肾小球、肾小管的形态规则、结构清晰、均匀分布，未见系膜细胞、内皮细胞增殖及纤维组织产生；DN组大鼠的肾组织肾小球结构紊乱，细胞数量增多，体积增加，系膜细胞增生，系膜区扩大、变形，基底膜弥漫性增厚，肾小管官腔增宽，线粒体肿胀；与DN组对比，LP组、EH组及EM组大鼠肾脏病理改变减轻；(3).SOCS-3、IGF-1表达情况：12周末，免疫组化及Western blot法测定，与NC组对比，DN组大鼠肾组织中SOCS-3的表达增加，IGF-1的表达显著升高(P0.05)，LP组、EH组及EM组较DN组大鼠肾组织SOCS-3表达明显增多、IGF-1表达明显下降(P0.05)。结论：芡实可减少尿蛋白，保护肾功能。芡实可能通过上调SOCS-3的表达，抑制IGF-1的过表达，从而延缓DN进展，发挥对肾脏保护作用。
[Abstract]:Objective: to observe the effect of Euryale coryale on the expression of SOCS-3 and IGF-1 in diabetic nephropathy (DN) rats, and to explore the possible mechanism of delaying the progression of diabetic nephropathy. Methods: 60 healthy and clean male Sprague-Dawley rats were selected as normal control group (NC group) by random digital table method after adapting to the environment for one week. The other 50 rats were treated with single dose injection of STZ45mg / kg) to establish diabetic rat model. They were randomly divided into diabetic nephropathy group (DN group), diabetic nephropathy treated group (EH) with low dose cornstarch seed (DM) group (1.5 g / kg / d), diabetic nephropathy treated group (EM group) (3.0 g / kg 路kg / d), diabetic nephropathy treated group with high dose Euryale seed solid treatment group (6.0 g kg 路kg / d) and diabetic nephropathy (DN). Losartan potassium treatment group and LP group 30 mg / kg / d, normal group intraperitoneal injection of the same volume of sodium citrate buffer without STZ. In the model group, the blood glucose was measured 72 hours later. If the blood glucose value was 16.7 mmol / L, the 24h urine protein was measured 3 weeks after the establishment of the diabetic model, and the DN model was considered to be successful. Each experimental group was treated with the corresponding dose of drugs, and the model group and the control group were given the same volume of distilled water for 12 weeks. The body weight of rats in each group was recorded at the end of the 12th week of treatment. Urine samples and fasting blood samples were collected for 24 hours, respectively, to detect 24 hour urinary protein quantification, serum urea nitrogen bun and serum creatinine, and then intraperitoneal injection of chloral hydrate was used to anesthetize the rats. The abdominal cavity was opened layer by layer, blood was taken from the abdominal aorta, and the kidney was washed to gray-yellow. The pathological changes of renal tissue were observed by HE staining Masson staining and electron microscope. Immunohistochemical method and Western blot technique were used to detect the expression and distribution of SOCS-3 and IGF-1 in rat kidney. The result is that one is one. Comparison of biochemical parameters: blood glucose, blood urea nitrogen, serum creatinine 24 hours urine protein in NC group fluctuated within normal range at the end of 12 weeks, and blood urea nitrogen in DN group was higher than that in NC group. Compared with DN group, serum urea nitrogen and creatinine 24 hours urine protein in EM group and LP group were significantly decreased compared with DN group. At the end of 12 weeks, the glomeruli and renal tubules in the NC group were found to have regular morphology, clear structure, uniform distribution and no Mesangial cells. In the DN group, the glomerular structure of renal tissue in the group of endothelial cell proliferation and fibrous tissue formation was disordered, the number of cells increased, the volume increased, the Mesangial cell proliferated, the Mesangial area expanded, the basement membrane became diffuse thickening, and the renal tubule cavity widened. Compared with DN group, the pathological changes of kidney in LP group, EH group and EM group were reduced. The expression of SOCS-3 IGF-1 was reduced at the end of 12 weeks, and the expression of IGF-1 was determined by immunohistochemistry and Western blot method. Compared with NC group, the expression of SOCS-3 in renal tissue of DN group was significantly increased. The expression of IGF-1 was significantly increased in EH group and EM group than that in DN group. The expression of IGF-1 in kidney tissue of DN group decreased significantly (P 0.05). Conclusion: Euryale can reduce urine protein and protect renal function. Euryale Euryale may inhibit the overexpression of IGF-1 by upregulating the expression of SOCS-3, thus delaying the progression of DN and exerting its protective effect on kidney.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R587.2;R692

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