瑞芬太尼预处理对鼠肝缺血再灌注损伤时肾细胞凋亡及Bcl-2、caspase-3的影响

[目的]探讨瑞芬太尼预处理对大鼠肝脏缺血再灌注(IR)时远隔脏器肾脏细胞凋亡及 Bcl-2、半胱天冬酶家族3(caspase-3)的影响.[方法]健康雄性 SD 大鼠72只,随机分为假手术组(S组)、缺血再灌注组(IR组)、瑞芬太尼组(RPC 组),每组24只.采用Pringle''s maneuver法建立肝IR模型,检测各组缺血30 min(T1)、再灌注时间1 h(T2)、3 h(T3)、6 h(T4)时血清谷丙转氨酶(ALT)、肌酐(Cr)水平,再灌6 h取肾组织光镜观察肾脏病理损伤,同时TUNEL法检测肾脏细胞的凋亡,Western blot检测各组肾组织中B淋巴细胞瘤-2(Bcl-2)和caspase-3蛋白的表达水平.[结果]IR组、RPC组从缺血30 min开始各时点血清ALT、Cr依次升高,均显著高于S组,但RPC组又明显低于IR组,其差异均有统计学意义(P<0.05).S组大鼠肾组织结构正常;IR组可见肾脏组织不同程度的病理损伤:肾小管上皮细胞肿胀、结构欠清、核浓缩、肾小管内有上皮坏死脱落细胞的形成,间质炎性细胞浸润及充血;RPC组损伤明显改善.S组仅有极少量的凋亡阳性细胞,IR组、RPC组凋亡阳性细胞均显著高于S组(P<0.05),且IR组明显高于RPC组,其差异均有统计学意义(P<0.05).与S组比较,肾组织Bcl-2、caspase-3表达水平显著增高(P<0.05);与IR组比较,RPC组Bcl-2水平增高(P<0.05),caspase-3表达量明显下降(P<0.05).[结论]瑞芬太尼对肝IR时肾脏损伤有保护作用,而其保护机制可能与调控肾细胞凋亡和Bcl-2、Caspase-3蛋白的表达有关.

Effect of Remifentanil Preconditioning on the Apoptosis of Renal Cells and Expression of Bcl-2 and Caspase3 after Hepatic Ischemia Reperfusion in Rats

[Objective]To evaluate the effect of remifentanil preconditioning on the apoptosis of renal cells and expressions of Bcl-2 and Caspase3 after hepatic ischemia reperfusion in rats.[Methods]Seventy-two adult male SD rats were randomly divided into three equal groups (n=24): the sham operation group (group S),ischemia reperfusion group (group I/R),and remifentanil group (group RPC).According to 4 different time points of ischemia and reperfusion [ischemia for 30 min (T1),reperfusion for 1h (T2),3h (T3) and 6h (T4)],the rats were further subdivided into 4 subgroups within each group: S1,S2,S3,S4;IR1,IR2,IR3,IR4,;and RPC1,RPC2,RPC3,RPC4.The Hepatic ischemia reperfusion model in rats was established using Pringle's maneuver method.The serum levels of alanine aminotransferase (ALT) and creatinine (Cr) were examined at each time point mentioned above.After 6 hours of reperfusion,the pathological changes in the kidneys were observed.TUNEL staining was used to examine cell apoptosis,and Western blot was employed to examine protein expressions of Bcl-2 and Caspase3 in renal tissues of each group.[Results]Serum ALT and creatinine levels in the I/R group and RPC group increased after 30 min of ischemia-these changes were significantly higher than that in the S group (P<0.05).However,levels in RPC group were lower than those in the I/R group (P<0.05).The renal tissue structure of the rats in group S was normal.In the IR group,there were different degrees of pathological damage of renal tissue: renal tubular epithelial cell swelling,lack of structure,nucleus condensation,necrotic exfoliated epithelial cells,and inflammatory cell infiltration.The pathological damage was obviously improved following remifentanil preconditioning in RPC group.TUNEL staining revealed that there were significantly lower apoptotic cells in the S group than those in the I/R group and the RPC group (P<0.05),and that the degree in the I/R group was much higher than that in the RPC group (P<0.05).Western blot detection of apoptotic protein showed that the expression of Bcl-2 and Caspase3 in the I/R group were higher than those in the S group (P<0.05).After Remifentanil preconditioning in RPC group,the expression of Bcl-2 increased but Caspase-3 expression decreased (P<0.05).[Conclusion]Remifentanil preconditioning has a protective effect on renal injury after hepatic ischemia reperfusion,and the protective mechanism may be involved in the regulation of renal cell apoptosis and the expression of Bcl-2 and Caspase-3.