瑞芬太尼对大鼠肾缺血再灌注时TLR2 mRNA表达的影响

Effect of remifentanil on Toll-like receptor 2 mRNA expression during renal ischemia-reperfusion in rats

目的 探讨瑞芬太尼对大鼠肾缺血再灌注时Toll样受体2（TLR2） mRNA表达的影响.方法 健康成年雄性SD大鼠54只,体重200～ 250 g,采用随机数字表法分为3组（n=18）：假手术组（S组）、肾缺血再灌注组（Ⅰ/R组）和瑞芬太尼组（R组）.Ⅰ/R组和R组采用夹闭双侧肾动脉45min恢复再灌注的方法制备大鼠肾缺血再灌注损伤模型.R组于缺血前15 min至再灌注30 min经尾静脉输注瑞芬太尼1.0 μg· kg-1·min-1,S组和Ⅰ/R组输注等容量生理盐水.于缺血前15 min、再灌注6和24 h时取肾组织,光镜下观察肾组织病理学结果,采用ELISA法检测肾组织TNF-α和IL-6的含量,流式细胞术双染法测定细胞凋亡率,RT-PCR法检测TLR2 mRNA的表达水平.结果 与S组比较,Ⅰ/R组和R组再灌注6和24 h时肾组织TLR2 mRNA表达上调,TNF-α和IL-6含量及细胞凋亡率升高（P＜0.01）;与Ⅰ/R组比较,R组再灌注6和24 h时肾组织TLR2 mRNA表达下调,TNF-α和IL-6含量及细胞凋亡率降低（P＜0.05或0.01）.R组肾组织病理学损伤较Ⅰ/R组减轻.结论 瑞芬太尼减轻大鼠肾缺血再灌注损伤的机制与其下调TLR2表达,降低其活性,进而抑制肾组织炎性反应和细胞凋亡有关.

Objective To evaluate the effect of remifentanil on Toll-like receptor 2 （TLR2） mRNA expression during renal ischemia-reperfusion （Ⅰ/R） in rats.Methods Fifty-four male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 3 groups （n=18 each） using a random number table：sham operation group （group S）,Ⅰ/R group and remifentanil group （group R）.Renal Ⅰ/R injury was produced by clamping the bilateral renal arteries for 45 min followed by reperfusion in Ⅰ/R and R groups.Bilateral renal arteries were only exposed but not clamped in group S.Remifentanil 1.0 μg · kg-1 · min-1 was infused via the tail vein starting from 15 min before ischemia until 30 min of reperfusion in group R,while the equal volume of normal saline was given instead in S and Ⅰ/R groups.The animals were sacrificed at 15 min before ischemia and 6 and 24 h of reperfusion,and the renal specimens were obtained for examination of the pathological changes （with light microscope） and for determination of the contents of tumor necrosis factor-alpha （TNF-α） and interleukin-6 （IL-6） （by ELISA） and expression of TLR2 mRNA （by RT-PCR） and cell apoptosis （by double staining and flow eytometry）.The apoptotic rate was calculated.Results Compared with group S,TLR2 mRNA expression was significantly up-regulated,and the contents of TNF-α and IL-6 and apoptotic rate were increased at 6 and 24 h of reperfusion in Ⅰ/R and R groups.Compared with group Ⅰ/R,TLR2 mRNA expression was significantly down-regulated,and the contents of TNF-α and IL-6 and apoptotic rate were decreased at 6 and 24 h of reperfusion in group R.The pathological changes were significantly attenuated in group R as compared with group Ⅰ/R.Conclusion The mechanism by which remifentanil reduces renal Ⅰ/R injury is related to down-regulation of TLR2 expression and decrease in TLR2 activity and inhibition of inflammatory responses in renal tissues and cell apoptosis in rats.