本文選題：NALP3 + MAPK��； 參考：《山西醫(yī)科大學》2012年碩士論文

【摘要】：目的通過大鼠腎臟組織局部轉染NALP3特異性小干擾質粒（siRNA）后制作腎臟缺血再灌注損傷（IRI）模型，探討病原體相關分子構象識別受體NALP3對大鼠腎臟IRI的影響及其作用機制。 方法(1)將健康SD大鼠隨機分為假手術（sham）組、缺血再灌注（I/R）不同時間1h、2h、4h、8h、16h、24h組、空質粒+sham組、空質粒+I/R組和siRNA質粒+I/R組，摘除右腎一周后夾閉左腎動脈制作腎臟IRI模型，轉染組摘除右腎后超聲微泡造影技術完成轉染一周后同樣方法制作IRI模型，各組10大鼠，留取血清及腎組織標本。(2)對腎臟組織進行病理損傷半定量分析；檢測血清Scr、BUN含量的變化。(3) western印跡法半定量分析腎組織NALP3的表達變化、ERK、JNK、P38磷酸化程度變化；RT PCR檢測NALP3mRNA表達變化。 結果(1)大鼠缺血再灌注不同時間均發(fā)生不同程度急性腎小管壞死，各組病理評分中以缺血再灌注4h組病理損傷最重（P＜0.01）。(2)大鼠缺血再灌注不同時間血清Scr、BUN水平均有不同程度升高，以4h組最高。(3)與假手術組比較，NALP3在缺血再灌注1h、2h、4h、8h、16h、24h表達顯著增高（P＜0.05），4h達到最高，之后趨于穩(wěn)定。(4)與單純缺血再灌注4h組比較，，siRNA質粒+I/R組腎組織病理損傷明顯減輕，JNK、P38磷酸化程度明顯下降(P＜0.01)，ERK磷酸化程度仍然較高。 結論1)NALP3在大鼠腎臟缺血再灌注損傷中表達增加，下調其表達后腎臟缺血再灌注損傷明顯改善。2) MAPK通路在大鼠腎臟缺血再灌注損傷中磷酸化程度明顯增強，下調NALP3后磷酸化程度明顯減少，提示下調NALP3對大鼠腎臟缺血再灌注損傷有明顯保護作用，并且是通過抑制JNK、P38的活化實現的。
[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.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R363

【參考文獻】

相關期刊論文 前1條

1 姜勇,龔小衛(wèi);MAPK信號轉導通路對炎癥反應的調控[J];生理學報;2000年04期

本文編號：2096601