雷公藤内酯醇对肾缺血再灌注大鼠凋亡诱导因子及细胞间黏附分子-1表达的影响

Effects of Triptolide on Expression of AIF and ICAM-1 in Rat's Kidney Tissure with Renal Ischemia Reperfusion Injury

目的:探讨雷公藤内酯醇(TP)对肾缺血再灌注(I/R)大鼠的肾保护作用,及对凋亡诱导因子、细胞间黏附分子-1的影响。方法:48只雄性Wistar大鼠随机分为假手术组、I/R模型组(I/R组)、TP高、中、低剂量干预组、泼尼松对照组(Pred组)。采用夹闭双侧肾动脉30 min,再灌注18 h的方法制作肾I/R大鼠模型。检测血肌酐(Scr)、尿素氮(BUN),原位末端标记法检测肾小管上皮细胞凋亡,观察肾病理改变,计算肾小管损伤(ATN)评分。Western印迹和RT-PCR分别检测AIF、ICAM-1蛋白和基因表达。结果:(1)I/R组血Scr、BUN、ATN评分及细胞凋亡指数较假手术组显著升高(P<0.01);与I/R组比较,TP各组以上指标均改善(P<0.01),pred组血Scr、BUN、ATN评分改善(P<0.01),但细胞凋亡指数无明显变化(P>0.05)。(2)I/R组大鼠肾组织AIF、ICAM-1较假手术组高表达(P<0.01);与I/R组比较,TP各组二者表达均减弱(P<0.01),pred组ICAM-1表达减弱(P<0.01)但AIF表达差异无统计学意义(P>0.05)。结论:TP对肾I/R大鼠具有肾保护作用,其部分机制可能与抑制肾小管上皮细胞过度凋亡及AIF、ICAM-1表达有关。

Objective： To explore protective effect of Triptolide （TP） on rat＇ s kidney from renal ischemia reperfusion （ 1/ R） injury and interference effect on expression level of apoptosis - inducing factor （AIF） and intercellular adhesion molecule - 1 and （ ICAM - 1 ）. Methods ：48 male Wistar rats were randomly divided into six groups ： sham operation group, ischemia reperfusion injury group （I/R group）, low TP group, medium TP group, high TP group, prednisone group （Pred group）. The renal I/R rat model was duplicated by clamping Bilateral renal artery for 30 minutes and then artery reperfusion for 18 hours. The levels of serum creatinine （Scr） and Blood urea nitrogen （Bun） were detected. Apoptosis of tubular epithelial cell was determined by terminal deoxynucleotidy transferase dUTP nick end labeling （TUNEL）. Histological changes of kidney were observed and damage score of renal tubule were computed. The expression of AIF/ICAM - 1 protein and mRNA were detected by Western blotting and real - time PCR respectively. Results：（ 1 ）Scr, BUN, ATN score and the apoptotic index in I/R group were significantly increased compared to sham operation group （P 〈0.01 ）. Compared with I/R group them were significantly reduced in all riP groups （P 〈0.01 ） ,Scr, BUN and ATN score were significantly reduced in pred group （ P 〈 0.01 ）, but the apoptotie index not significantly difference in pred group （ P 〉 0.05 ）. （2） The expression of ICAM - 1 and AIF in I/R group were significantly up - regulated compared to sham operation group （ P 〈 0.01 ）. Compared with I/R group them were significantly down- regulated in all TP groups（P 〈0.01 ）, The expression of ICAM -1 was significantly down - regulated in pred group （ P 〈 0.01 ） , but the expression of AIF was simil level in pred group （ P 〉 0.05 ）. Conclusion：TP has protective effect on kidney from renal ischemia reperfusion injury. The mechanism maybe was associated with the down - regulated the apoptosis of renal tubule epithelial cell and the expressions of AIF and ICAM - 1.