血红素氧合酶-1通过p38MAPK途径抗老龄肾脏缺血再灌注损伤大鼠模型细胞凋亡作用的研究

发布时间：2018-07-01 15:31

  本文选题：缺血再灌注损伤 + 老年大鼠　； 参考：《中南大学》2012年硕士论文

【摘要】：目的： 探讨血红素氧合酶-1(Heme oxygenase-1)对老年大鼠缺血再灌注损伤(ischemia/reperfusion injury, IRI)'肾脏的抗凋亡作用及其机制。 方法： 25月龄健康雄性Wistar大鼠随机分为正常对照组、I/R组、CoPP组、ZnPP组和CoPP+SB203580组。成功建模后24h,取肾脏组织,分别用RT-PCR和Western blot检测HO-1蛋白的表达；免疫组化检测Caspase-3的表达；流式细胞术检测肾组织细胞凋亡情况；Western blot检测P38MAPK的磷酸化的情况。 结果： RT-PCR和Western blot结果显示：对照组肾组织中存在HO-1分子的mRNA和蛋白的表达；与对照组相比,I/R组HO-1的表达明显增加,HO-1诱导剂CoPP增加了HO-1的表达,HO-1抑制制剂ZnPP抑制了HO-1的表达。免疫组化结果显示I/R组Caspase-3表达水平明显升高,与I/R组比较,CoPP组肾组织Caspase-3表达水平显著降低,而ZnPP组和CoPP+SB203580组Caspase-3表达水平￡显著升高。流式细胞术结果显示,I/R组细胞凋亡率明显升高,与I/R组比较,CoPP组细胞凋亡率显著降低,而ZnPP组和CoPP+SB203580组细胞凋亡率明显升高；Western blot结果显示I/R组p38MAPK的磷酸化水平明显升高,HO-1诱导剂CoPP促进了p38MAPK的磷酸化,HO-1抑制剂ZnPP抑制了p38MAPK的磷酸化,SB203580同样抑制了p38MAPK的磷酸化。 结论： HO-1对老年大鼠缺血再灌注损伤的保护作用可能是通过激活P38MAPK的磷酸化,抑制凋亡因子的表达,减少细胞凋亡,从而减轻肾脏损伤及功能障碍来实现。
[Abstract]:Aim: to investigate the antiapoptotic effect of heme oxygenase-1 on ischemia/reperfusion injury-reperfusion injury (IRI) 'kidney in aged rats and its mechanism. Methods: healthy male Wistar rats aged 25 months were randomly divided into two groups: normal control group (I / R group) and Copp group (ZnPP group) and CoPP SB203580 group. The expression of HO-1 protein was detected by RT-PCR and Western blot, the expression of Caspase-3 was detected by immunohistochemistry, the apoptosis of renal tissue was detected by flow cytometry and the phosphorylation of P38 MAPK was detected by Western blot. Results: the results of RT-PCR and Western blot showed that the expression of HO-1 mRNA and protein existed in the control group. Compared with the control group, the expression of HO-1 in IPA / R group was significantly increased, and the expression of HO-1 was increased by CoPP, the inducer of HO-1, and the expression of HO-1 was inhibited by ZnPP, an inhibitor of HO-1. The results of immunohistochemistry showed that the expression of Caspase-3 was significantly increased in I / R group. Compared with I / R group, the expression level of Caspase-3 in renal tissue of Caspase-3 was significantly decreased, while that of Caspase-3 in ZnPP group and Copp SB203580 group was significantly higher than that in I / R group. The results of flow cytometry showed that the apoptosis rate of I- / R group was significantly higher than that of I- / R group, but the apoptosis rate of ZnPP group and Copp SB203580 group was significantly higher than that of I / R group. Western blot showed that the phosphorylation level of p38 MAPK was significantly increased in I / R group. Copp promoted the phosphorylation of p38 MAPK and inhibited the phosphorylation of p38 MAPK, SB203580 and p38 MAPK, as well as the phosphorylation of p38 MAPK. Conclusion: the protective effect of HO-1 on ischemia-reperfusion injury in aged rats may be achieved by activating phosphorylation of p38 MAPK, inhibiting the expression of apoptotic factor and reducing apoptosis, thereby reducing renal injury and dysfunction.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R-332;R692

【参考文献】

相关期刊论文 前3条

1 魏金星,武玉东,刘秉乾,杨俊福,高建光;氯化亚锡对大鼠肾缺血再灌注损伤的作用和血红素氧合酶-1表达的影响[J];郑州大学学报(医学版);2005年05期

2 赵学义,李国毅,何明艳,徐勇杰,李德谦,郭跃虎,田晋洪,宁林红;55岁以上活体亲属供肾移植相关问题探讨[J];中华外科杂志;2003年12期

3 周成;谢晋良;朱向荣;丁翔;王耀磊;;重组腺相关病毒介导的HO-1基因转染对大鼠移植肾缺血再灌注损伤的保护作用[J];中国现代医学杂志;2010年18期

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