熊果酸对阿霉素肾病的干预作用研究

发布时间：2018-05-14 10:08

本文选题：熊果酸 + 阿霉素肾病　；参考：《福建医科大学》2014年硕士论文

【摘要】：目的研究熊果酸对小鼠阿霉素肾病的影响，探讨其可能的机制，为防止或延缓肾纤维化发展提供实验依据，同时为临床防治肾纤维化寻求新的药物。方法将雄性Balb/c小鼠随机分为4组，，即正常对照组（Normal组）、阿霉素肾病组（ADN组）、低剂量熊果酸干预组（LU+ADN组）、高剂量熊果酸干预组（HU+ADN组），每组10只。实验第4天，ADN组、LU+ADN组、HU+ADN组小鼠经尾静脉单次注射ADR10mg/Kg创建ADN模型，Normal组予等量生理盐水。实验第1~18天，LU+ADN组、HU+ADN组分别予UA25mg/（Kg·d）、50mg/（Kg·d）腹腔注射，Normal组、ADN组予等量溶媒（14%DMSO）。造模后7天，收集24小时尿液，测定尿蛋白总量。造模后14天处死，留取血清检测SCr、BUN、MDA，HE染色观察肾脏病变情况，免疫组织化学法检测TGF-β1在肾小管间质中的表达，进行半定量分析。结果1.一般情况:造模后，ADN组小鼠状态略差，进食、活动等有所减少，精神稍萎靡，LU+ADN组及HU+ADN组小鼠状态无明显变化。Normal组小鼠体重逐渐增长。各组小鼠造模后，体重均明显下降，不同剂量的UA对体重变化无明显影响。2.小鼠24小时尿蛋白总量:ADN组小鼠较Normal组显著升高（P＜0.05），HU+ADN组较ADN组显著下降（P＜0.05）。3.肾功能：各组小鼠SCr、BUN差异均无统计学意义。4.血清MDA：注射ADR的三组小鼠较Normal组显著升高（P＜0.01），两治疗组MDA水平与ADN组无显著差异。5.HE染色: Normal组小鼠肾组织正常。ADN组肾脏表现为局灶性节段性肾小球硬化，肾小管上皮细胞肿胀，部分缺如，肾小管明显扩张，管腔内充满均质红染的蛋白管型，间质形成纤维化病变；与ADN组相比，LU+ADN组肾脏病变改善不明显，HU+ADN组肾脏病变较轻。6.免疫组化染色半定量分析:①与Normal组比，ADN组肾间质表达TGF-β1显著增加（P＜0.05）。②与ADN组比，LU+ADN组TGF-β1表达差异无统计学意义；HU+ADN组小鼠TGF-β1表达显著下降（P＜0.05）。结论1、证实高剂量熊果酸（50mg/Kg）可以减少阿霉素肾病小鼠的尿蛋白。2、证实高剂量熊果酸可以抑制阿霉素肾病小鼠肾小管间质TGF-β1蛋白的表达，抑制或延缓肾小球硬化、肾小管间质纤维化进展，发挥肾脏保护作用。3、本实验为熊果酸作为降低尿蛋白、抗肾纤维化新的药物开发提供了实验依据。
[Abstract]:Objective to study the effect of ursolic acid on adriamycin nephropathy in mice and explore its possible mechanism in order to provide experimental basis for preventing or delaying the development of renal fibrosis and to seek new drugs for clinical prevention and treatment of renal fibrosis. Methods male Balb/c mice were randomly divided into 4 groups: normal control group, adriamycin nephropathy group, low dose ursolic acid intervention group and high dose ursolic acid intervention group. On the 4th day of the experiment, the mice in the adn group were injected with ADR10mg/Kg through the tail vein to create the ADN model. The normal group was given the same amount of normal saline. The mice in the LU ADN group were given the same amount of normal saline by single injection of ADR10mg/Kg through the tail vein. In the first 18 days of the experiment, Hu ADN group was given 50 mg / g / kg UA25mg/(Kg dago in the ADN group, respectively) and the normal group was given the same amount of solute and 14DMSOO in the normal group. After 7 days, 24 hours urine was collected to determine the total amount of urine protein. The rats were killed 14 days after the model. The serum samples were collected to detect the pathological changes of kidney by SCrBUNMDAHE staining, and the expression of TGF- 尾 1 in renal tubulointerstitium was detected by immunohistochemical method, and the expression of TGF- 尾 1 in renal tubulointerstitium was detected by semi-quantitative analysis. Result 1. General conditions: after modeling, the mice in the ADN group were in poor state, eating and activities were decreased, and the mental state of the LU ADN group and Hu ADN group had no obvious change. The body weight of the normal group gradually increased. The body weight of each group of mice decreased significantly after modeling, and the changes of body weight were not affected by UA at different doses. Compared with Normal group, the total urine protein content in 24 hour group was significantly higher than that in Normal group (P < 0.05). The content of urine protein in ADN group was significantly lower than that in ADN group (P < 0.05). Renal function: there was no significant difference of SCrBUN in each group. 4. 4. Serum MDA: compared with the Normal group, the MDA level of the three groups was significantly higher than that of the Normal group (P < 0.01). There was no significant difference between the two groups in the level of MDA. 5. He staining showed that the renal tissue of the Normal group was normal. The kidney of the Normal group showed focal segmental glomerulosclerosis and tubule epithelial cells swelling. The renal tubules were dilated obviously, the tubules were filled with homogeneous red staining protein tubules, and the interstitial fibrosis lesions were formed in the tubules, and the renal lesions in the LU ADN group were not significantly improved compared with those in the ADN group, and the renal lesions in the Hu ADN group were less than that in the Hu ADN group. The expression of TGF- 尾 _ 1 in renal interstitial tissue of Normal group was significantly increased by immunohistochemical staining than that of Normal group (P < 0.05). There was no significant difference in TGF- 尾 _ 1 expression between LU ADN group and ADN group. The expression of TGF- 尾 _ 1 in HU ADN group was significantly lower than that in HU ADN group (P < 0.05). Conclusion 1. It is proved that high dose ursolic acid 50 mg / Kg can reduce urinary protein 0.2 in adriamycin nephropathy mice, and that high dose ursolic acid can inhibit the expression of TGF- 尾 1 protein in renal tubulointerstitial of adriamycin nephropathy mice and inhibit or delay glomerulosclerosis. The progress of renal tubulointerstitial fibrosis (tubulointerstitial fibrosis) plays a protective role in kidney. This experiment provides experimental basis for the development of ursolic acid as a new drug to reduce urinary protein and resist renal fibrosis.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692

【参考文献】