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目的异氟醚预处理能对大鼠肾脏发挥急性期缺血保护作用,但急性期保护作用仅能维持2～3h。探讨异氟醚延迟性预处理是否能减轻肾脏缺血再灌注损伤,以及缺氧诱导因子1α(hypoxia inducible factor1α,HIF-1α)是否参与调节该保护作用。方法取雄性C57BL/6小鼠52只,体重20～25g,随机分为4组(n=13):对照组(A组)、PBS加异氟醚预处理组(B组)、无意义小干扰RNA(small interference RNA,siRNA)加异氟醚预处理组(C组)和HIF-1αsiRNA加异氟醚预处理组(D组)。气体暴露前24h C、D组小鼠经尾静脉分别给予含有50μg无意义siRNA或HIF-1αsiRNA的无RNA酶的PBS1mL;A、B组给予等体积PBS。B、C、D组给予含1.5%异氟醚和25%O2的气体持续吸入2h,A组仅吸入含25%O2的气体2h。气体暴露后24h,采用Western blot技术检测肾脏组织中HIF-1α和促红细胞生成素(erythropoietin,EPO)蛋白的表达;制备肾缺血再灌注损伤,于再灌注后24h检测血清肌酐(serum creatinine,SCr)和血尿素氮(blood urea nitrogen,BUN)水平,并观察肾组织病理学变化。结果与A组比较,B、C组HIF-1α和EPO蛋白表达明显增加(P<0.01),SCr和BUN水平以及肾小管评分明显降低(P<0.01);D组HIF-1α蛋白表达及SCr、BUN水平较A组明显下降(P<0.01)。与B、C组比较,D组HIF-1α和EPO蛋白的表达明显下降(P<0.01),SCr和BUN水平以及肾小管评分明显增加(P<0.01),肾组织损伤明显加重。结论异氟醚延迟性预处理能明显减轻肾缺血再灌注损伤,HIF-1α参与调控异氟醚的保护作用。
Purpose Isoflurane preconditioning can exert the acute phase of ischemic injury in rat kidneys, but the protective effect in acute phase can only be maintained for 2 ~ 3h. To investigate whether prolonged isoflurane preconditioning can reduce renal ischemia / reperfusion injury and whether hypoxia inducible factor 1α (HIF-1α) is involved in the protective effect. Methods 52 male C57BL / 6 mice weighing 20-25g were randomly divided into 4 groups (n = 13): control group (group A), PBS plus isoflurane preconditioning group (group B) (SiRNA) plus isoflurane preconditioning (group C) and HIF-1α siRNA plus isoflurane preconditioning (group D). 24 h before gas exposure, mice in group D were given 1 mL of RNase-free PBS containing 50 μg of siRNA or HIF-1α siRNA via caudal vein. Groups A and B received equal volume of PBS. B, C and D groups were given inhaled gas containing 1.5% isoflurane and 25% O2 for 2h. Group A only inhaled 25% O2 gas for 2h. The expression of HIF-1|Á and erythropoietin (EPO) in renal tissues was detected by Western blot 24h after gas exposure. Renal ischemia / reperfusion injury was induced and serum creatinine SCr) and blood urea nitrogen (BUN) were measured. The pathological changes of renal tissue were observed. Results Compared with group A, the expression of HIF-1α and EPO protein in group B and group C were significantly increased (P <0.01), while the levels of SCr and BUN and tubule score were significantly decreased (P <0.01). The protein expression of HIF-1α and SCr , BUN level was significantly lower than that of A group (P <0.01). Compared with group B and group C, the expression of HIF-1α and EPO in group D was significantly decreased (P <0.01), and the levels of SCr and BUN and tubule score were significantly increased (P <0.01). Conclusions Delayed preconditioning with isoflurane can significantly reduce renal ischemia-reperfusion injury, and HIF-1α is involved in the regulation of the protective effect of isoflurane.