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

摘要

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 TP groups ( P <0. 01 ) ,Scr, BUN and ATN score were significantly reduced in pred group ( P < 0. 01 ), but the apoptotic 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.