NALP3在大鼠肾缺血再灌注损伤中的表达及作用研究

发布时间：2018-07-04 16:01

本文选题：NALP3 + MAPK　；参考：《山西医科大学》2012年硕士论文

【摘要】：目的通过大鼠肾脏组织局部转染NALP3特异性小干扰质粒（siRNA）后制作肾脏缺血再灌注损伤（IRI）模型，探讨病原体相关分子构象识别受体NALP3对大鼠肾脏IRI的影响及其作用机制。方法(1)将健康SD大鼠随机分为假手术（sham）组、缺血再灌注（I/R）不同时间1h、2h、4h、8h、16h、24h组、空质粒+sham组、空质粒+I/R组和siRNA质粒+I/R组，摘除右肾一周后夹闭左肾动脉制作肾脏IRI模型，转染组摘除右肾后超声微泡造影技术完成转染一周后同样方法制作IRI模型，各组10大鼠，留取血清及肾组织标本。(2)对肾脏组织进行病理损伤半定量分析；检测血清Scr、BUN含量的变化。(3) western印迹法半定量分析肾组织NALP3的表达变化、ERK、JNK、P38磷酸化程度变化；RT PCR检测NALP3mRNA表达变化。结果(1)大鼠缺血再灌注不同时间均发生不同程度急性肾小管坏死，各组病理评分中以缺血再灌注4h组病理损伤最重（P＜0.01）。(2)大鼠缺血再灌注不同时间血清Scr、BUN水平均有不同程度升高，以4h组最高。(3)与假手术组比较，NALP3在缺血再灌注1h、2h、4h、8h、16h、24h表达显著增高（P＜0.05），4h达到最高，之后趋于稳定。(4)与单纯缺血再灌注4h组比较，，siRNA质粒+I/R组肾组织病理损伤明显减轻，JNK、P38磷酸化程度明显下降(P＜0.01)，ERK磷酸化程度仍然较高。结论1)NALP3在大鼠肾脏缺血再灌注损伤中表达增加，下调其表达后肾脏缺血再灌注损伤明显改善。2) MAPK通路在大鼠肾脏缺血再灌注损伤中磷酸化程度明显增强，下调NALP3后磷酸化程度明显减少，提示下调NALP3对大鼠肾脏缺血再灌注损伤有明显保护作用，并且是通过抑制JNK、P38的活化实现的。
[Abstract]:Objective to investigate the effects of NALP3 specific small interference plasmid (siRNA) on renal ischemia reperfusion injury (IRI) and its mechanism. Methods (1) healthy Sprague-Dawley rats were randomly divided into sham-operated (sham) group, ischemia reperfusion (I / R) group, empty plasmid sham group, empty plasmid I / R group and siRNA plasmid I / R group. In the transfection group, the IRI model was made one week after transfection with the right post-renal ultrasound microbubble technique. Serum and renal tissue samples were collected from 10 rats in each group. (2) Semi-quantitative analysis of renal injury was carried out. (3) the changes of NALP3 expression in renal tissue were detected by western blotting. (3) the changes of phosphorylation degree of ERKN JNKP P38 in renal tissue were detected by RT-PCR and the expression of NALP3 mRNA was detected by reverse transcription-polymerase chain reaction (RT PCR). Results (1) Acute tubular necrosis occurred at different time after ischemia reperfusion in rats. In the pathological score of each group, the pathological injury was the most serious in the 4 h ischemia reperfusion group (P < 0. 01). (2). The level of serum Scr-bun was increased in different time after ischemia reperfusion in rats. The highest expression of NALP3 was observed in the 4h group (P < 0.05). (3) the expression of NALP3 in the ischemia reperfusion group was significantly higher than that in the sham operation group (P < 0.05), and the expression of NALP3 was significantly increased at 1h, 2h, 4h, 8h, 16h and 24h (P < 0.05). (4) compared with the ischemia reperfusion group for 4 h, the pathological damage of renal tissue in the siRNA plasmid I / R group was significantly reduced (P < 0.01), and the phosphorylation degree of P38 was still higher in the siRNA plasmid I / R group than in the ischemia reperfusion group (P < 0.01). Conclusion 1) the expression of NALP3 in renal ischemia-reperfusion injury was increased, and the renal ischemia-reperfusion injury was improved significantly after down-regulation of NALP3. 2) the phosphorylation of MAPK pathway in renal ischemia-reperfusion injury was significantly increased in rats. The decrease of phosphorylation after down-regulation of NALP3 suggests that down-regulation of NALP3 has an obvious protective effect on renal ischemia-reperfusion injury in rats, and it is achieved by inhibiting the activation of JNKP38.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363

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