过度内质网应激在大鼠慢性环孢素A肾毒性模型细胞凋亡中的作用机制

发布时间：2018-10-22 18:57

【摘要】：目的：探讨过度内质网应激在大鼠慢性环孢素A (Cy-closporine A, CsA)肾毒性模型细胞凋亡中的作用机制。方法：将Sprague-Dawley (SD)大鼠随机分为正常对照组和慢性CsA肾毒性组,分别给予皮下注射橄榄油(1mL/kg/d,对照组)和CsA (15mg/kg/d,毒性组)1周及4周后处死大鼠。通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法(TUNEL染色法)及Masson染色评估肾小管间质纤维化和细胞凋亡：使用免疫组化学染色和免疫印迹依次检测分子免疫球蛋白结合蛋白(Bip)、磷酸化的真核细胞翻译起始因子2α(elF2α)、生长抑制DNA损伤基因153(GADD15)、细胞凋亡相关基因caspase-12及caspase-3蛋白表达。结果：与对照组比较,CsA注射1周观察不到肾小管间质纤维化和TUNEL阳性细胞,而给予CsA注射4周大鼠表现为明显的肾小管间质纤维化[(38.9±3.3)%vs.(0.0±0.0)%,P0.01]和大量TUNEL阳性细胞[(89±9)%vs.(7±2)%,pO.01]。在分子水平上,与对照组比较,CsA注射组Bip和caspase-12蛋白的表达1周达到高峰,4周后降至正常水平；反之,elF2α、 GADD153、caspase-3蛋白表达呈时间依赖性的增加。结论：在慢性CsA肾毒性中,过度的内质网应激耗尽Bip,凋亡前途径中e1F2α、GADD153、caspase-12, caspase-3的激活导致肾小管上皮细胞凋亡参与肾小管间质损伤。
[Abstract]:Aim: to investigate the mechanism of excessive endoplasmic reticulum stress (ERS) on apoptosis of chronic cyclosporine A (Cy-closporine A, CsA) nephrotoxic model in rats. Methods: Sprague-Dawley (SD) rats were randomly divided into normal control group and chronic CsA nephrotoxicity group. The rats were killed after 1 week and 4 weeks of subcutaneous injection of olive oil (1 mL / kg / d, control group) and CsA (15 mg / kg / d, toxicity group) respectively. Evaluation of tubulointerstitial fibrosis and apoptosis by terminal deoxynucleotidyl transferase-mediated dUTP Nick end labeling (TUNEL staining) and Masson staining: molecular analysis using immunohistochemical staining and Western blotting Immunoglobulin binding protein (Bip),) phosphorylation of eukaryotic cell translation initiation factor 2 伪 (elF2 伪), growth inhibition of DNA damage gene 153 (GADD15), apoptosis-related genes caspase-12 and caspase-3 protein expression. Results: compared with the control group, tubulointerstitial fibrosis and TUNEL positive cells were not observed in rats injected with CsA for 1 week, but significant renal tubulointerstitial fibrosis (38.9 卤3.3)% vs., P 0.01) and TUNEL positive cells (89 卤9)% vs. (7 卤2)%, pO.01) were observed in rats treated with CsA for 4 weeks. At the molecular level, the expression of Bip and caspase-12 protein in CsA injection group reached the peak at 1 week and decreased to normal level after 4 weeks, whereas the expression of elF2 伪 and GADD153,caspase-3 protein increased in a time dependent manner. Conclusion: in chronic CsA nephrotoxicity, the activation of e1F2 伪 and GADD153,caspase-12, caspase-3 in the preapoptotic pathway of Bip, induced by excessive endoplasmic reticulum stress leads to the apoptosis of renal tubular epithelial cells involved in tubulointerstitial injury.
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692

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