缺血后处理对肾缺血再灌注损伤保护作用的基因表达谱研究

发布时间：2018-10-17 09:59

【摘要】：目的建立肾缺血再灌注损伤(ischemia-reperfusion injury, TRI)缺血后处理(ischemic postconditioning, IPO)大鼠模型,研究IPO对缺血再灌住损伤肾基因表达谱的影响,探讨其保护机制。方法选用8-12周成年健康SD雄性大鼠60只,随机分为6组：对照组(S组)、缺血再灌注组IR缺血后处理A、B、C、D组,通过测定血尿素氮(BUN)和肌酐(SCr),肾组织结构损伤程度评分,筛选出成功的IPO模型。然后选取成年健康SD雄性大鼠6只分为2组：缺血再灌注组和IPO组。缺血再灌注组：结扎右侧肾蒂,夹闭左侧肾蒂60min后恢复灌注。IPO组：在肾脏缺血60min后进行6次开放10s+阻断10s的循环然后充分开放灌注。饲养24h后迅速切取左肾,分别将左肾组织放入液氮低温保存。进行RNA抽提、探针制备和芯片杂交,杂交后立即进行扫描检测信号收集图像。应用Softfire Decisionsite8.0软件分析某基因是否有表达,以及两组表达程度是否有差异。根据基因编码蛋白的功能对差异表达超过两倍的基因进行初步分析。结果缺血后处理组与缺血再灌注组肾脏组织比较共有70条基因表达有差异,基因表达上调的有48条,基因表达下调的基因有22条。这些基因主要涉及细胞代谢与损伤、凋亡调控相关蛋白、信号转导和转录、炎症相关因子、氧化应激相关因子等。结论成功建立了肾缺血再灌注损伤IPO大鼠模型,IPO可能作用于多个基因靶点,减轻肾脏缺血再再灌注损伤。
[Abstract]:Objective to establish a model of renal ischemia-reperfusion injury (ischemia-reperfusion injury, TRI) in (ischemic postconditioning, IPO) rats, to study the effect of IPO on renal gene expression profile of ischemia-reperfusion injury and to explore its protective mechanism. Methods Sixty male adult SD rats aged 8-12 weeks were randomly divided into 6 groups: control group (S group) and ischemic reperfusion group (group D) treated with IR after ischemia. Blood urea nitrogen (BUN) and creatinine (SCr), (SCr),) were measured. The successful IPO model was screened out. Then 6 male adult SD rats were divided into two groups: ischemia reperfusion group and IPO group. In the ischemia-reperfusion group, the right renal pedicle was ligated, the left renal pedicle 60min was clipped and the perfusion was restored. In the IPO group, the circulation was blocked for 10 s for 6 times and then the perfusion was fully opened after 10 s of renal ischemia 60min. After feeding for 24 hours, the left kidney was quickly removed and stored in liquid nitrogen. RNA extraction, probe preparation and chip hybridization were performed immediately after hybridization. Softfire Decisionsite8.0 software was used to analyze whether a gene was expressed and whether there were differences between the two groups. According to the function of gene encoding protein, the differentially expressed genes were preliminarily analyzed. Results there were different expression of 70 genes in renal tissues between the post-ischemic group and the ischemia-reperfusion group. There were 48 up-regulated genes and 22 down-regulated genes. These genes are mainly involved in cell metabolism and injury, apoptosis-related proteins, signal transduction and transcription, inflammatory related factors, oxidative stress related factors and so on. Conclusion IPO rat model of renal ischemia-reperfusion injury was successfully established. IPO may act on multiple gene targets to alleviate renal ischemia-reperfusion injury.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692

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