IL-17介导原发性肾病综合征肾脏损害及槐杞黄肾脏保护作用的实验研究

发布时间：2018-04-17 15:13

本文选题：槐杞黄颗粒 + 阿霉素肾病　；参考：《重庆医科大学》2012年硕士论文

【摘要】：目的：观察炎症因子白介素-17（interleukin-17）在阿霉素肾病小鼠肾小球硬化进程中表达情况，探讨槐杞黄颗粒对小鼠阿霉素肾病的保护作用及可能机制。方法：雄性，8w，BAL b/c小鼠随机分为正常对照组、阿霉素肾病组和阿霉素肾病阿霉素肾病槐杞黄治疗组，每组12只。经尾静脉一次性注射10mg/kg阿霉素建立阿霉素肾病小鼠模型。阿霉素肾病槐杞黄治疗组自阿霉素注射后第7d起，每日给予4g/kg槐杞黄清膏灌胃，正常对照组和阿霉素肾病组每日给予等量蒸馏水灌胃，灌胃共7周。观察小鼠一般状况和体重改变。阿霉素注射后第2、8周末分别处死小鼠，每组各6只。光学显微镜下观察肾组织病理改变；免疫组化检测肾脏局部IL-17表达情况；考马斯亮蓝法检测24h尿蛋白含量；全自动生化分析仪检测血清白蛋白和肌酐水平。结果：实验前正常对照组、阿霉素肾病组、阿霉素肾病槐杞黄治疗组小鼠体重和24h尿蛋白含量无明显差别。阿霉素肾病组小鼠自实验第2周开始出现大量蛋白尿、血清白蛋白降低持续至实验结束（与正常对照组比较，P0.05）；实验第8周，肾组织IL-17表达明显增强，伴有明显的局灶节段性肾小球硬化，甚至肾小球硬化及肾功能不全（与正常对照组比较，P0.05）。与同时间点阿霉素肾病组相比，阿霉素肾病槐杞黄治疗组小鼠24h尿蛋白明显降低、血清白蛋白升高、肾组织病理改变及肾功能损害程度减轻（P0.05）。结论：槐杞黄可能通过降低阿霉素肾病小鼠肾组织IL-17表达，减轻肾脏局部炎症反应，发挥肾脏起保护作用。目的：探讨interleukin-17(IL-17)对人系膜细胞体外增殖、合成分泌细胞外基质的影响及槐耳清膏的干预作用。方法：体外培养的人系膜细胞分为正常对照组、 IL-17组、IL-17+槐耳清膏组、槐耳清膏对照组。台盼蓝拒染法检测槐耳清膏的细胞毒性，MTT法测系膜细胞增殖情况，ELISA检测细胞培养上清液中纤连蛋白（FN）水平。结果：（1）各实验浓度槐耳清膏均无明显细胞毒性作用；（2）与正常对照组比较，不同浓度IL-17均可促进系膜细胞增殖（P0.05），各浓度之间无明显差异；不同浓度的槐耳清膏可抑制IL-17诱导的系膜细胞增殖，与IL-17（10ng/ml）组比较有显著差异（P0.05）；（3）槐耳清膏可抑制IL-17促系膜细胞合成分泌FN的作用，且可能具有一定时间量效关系。结论: IL-17能够诱导人系膜细胞增殖，促进系膜细胞合成分泌纤连蛋白。槐耳清膏可能通过抑制IL-17的刺激作用，发挥肾脏保护功能，从而延缓肾小球硬化的进展。
[Abstract]:Objective: To observe the expression of inflammatory factor interleukin -17 (interleukin-17) in the glomerulosclerosis process of adriamycin nephropathy mice, and to explore the protective effect and possible mechanism of Huai Qi Huang Granule on adriamycin nephrosis in mice.
Methods: male, 8W, BAL b/c mice were randomly divided into normal control group, adriamycin nephropathy group and adriamycin adriamycin nephropathy Huaiqihuang treatment group, 12 rats in each group. Adriamycin nephropathy mouse model by tail vein injection of 10mg/kg adriamycin. Adriamycin nephropathy Huaiqihuang treatment group after adriamycin injection 7d, daily give 4g/kg Huaiqihuang creamwith gavage, the normal control group and adriamycin group daily given distilled water orally by gavage for 7 weeks. The general condition and body weight of mice was observed. After adriamycin injection the mice were sacrificed after 2,8 weeks, 6 rats in each group. The renal pathologic changes were observed under light microscope; immune immunohistochemical detection of IL-17 in kidney of expression; detection of 24h urinary protein content by Coomassie brilliant blue method; automatic biochemical analyzer to detect the serum albumin and creatinine levels.
Results: there was no significant difference in body weight and 24h urine protein between the normal control group, adriamycin nephrosis group, adriamycin nephrosis group and Huai Qi Huang group before treatment.
Test second weeks massive proteinuria, serum albumin decreased continuously until the end of the experiment (compared with normal control group, P0.05); the eighth week, the expression of IL-17 was significantly enhanced, focal segmental glomerular sclerosis accompanied by obvious, even glomerular sclerosis and renal insufficiency (compared with normal control group, P0.05). Compared with the same time point of adriamycin nephropathy group, adriamycin nephropathy Huaiqihuang treatment group were significantly lower 24h urinary protein, serum albumin increased, the pathological change of kidney tissue and renal function damage reduction (P0.05).
Conclusion: sophorum huyfcii may reduce the expression of IL-17 in renal tissue of adriamycin nephrotic mice, reduce the local inflammatory reaction of kidney and play the protective role of kidney.
Objective: To investigate the effect of interleukin-17 (IL-17) on the proliferation of mesangial cells in vitro, affect the synthesis and secretion of extracellular matrix and effects of PS-T.
Methods: in vitro cultured human mesangial cells were divided into normal control group, IL-17 group, IL-17+ PS-T group, PS-T control group. Trypan blue to detect cytotoxicity of PS-T staining method, measuring mesangial cell proliferation by MTT method, the supernatant of fibronectin in cultured ELISA cells (detection the level of FN).
Results: (1) the experimental concentration of PS-T had no obvious cytotoxic effect; (2) compared with the normal control group, different concentration of IL-17 can promote the proliferation of mesangial cells (P0.05), no significant difference between the concentration; different concentrations of Huaier cream can inhibit the proliferation of mesangial cells induced by IL-17, and IL-17 (10ng/ml) group had significant difference (P0.05); (3) PS-T inhibited IL-17 induced mesangial cell synthesis and secretion of FN, and may have a certain amount of time effect relationship.
Conclusion: IL-17 can induce the proliferation of mesangial cells, promote mesangial cell synthesis and secretion of fibronectin. PS-T through inhibition of IL-17 stimulation, to prevent renal function, thereby delaying the progression of glomerular sclerosis.

【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R285.5;R726.9

【参考文献】