盐酸戊乙奎醚预处理对大鼠肝脏缺血-再灌注损伤的影响

Effects of penehyclidine hydrochloride preconditioning on hepatic ischemia-reperfusion injury in rats

目的评价盐酸戊乙奎醚预处理对大鼠肝脏缺血-再灌注损伤（HIRI）的影响。方法健康雄性SD大鼠54只,体重230-250g,采用随机数字表法将其均分为三组：假手术组（S组）、HIRI组（HR组）和盐酸戊乙奎醚预处理组（PHC组）。S组剖腹后仅牵拉肝十二指肠韧带;HR组用无创血管夹钳夹供应肝中叶和左叶的门静脉和肝动脉分支,造成70%肝脏缺血模型,45min后松开血管钳;PHC组的手术步骤与HIRI相同,PHC组术前30min肌肉注射0.45mg/kg的盐酸戊乙奎醚。分别于再灌注2h（T1）、4h（T2）和24h（T3）时每组随机取6只大鼠经腹主动脉采血,测定血清转氨酶（ALT）、天门冬氨酸转氨酶（AST）水平,并采用酶联免疫吸附法（ELISA）测定血清TNF-α和IL-1β浓度。采血结束后取肝组织,采用免疫组化法检测内皮型一氧化氮合酶（eNOS）及缺氧诱导因子（HIF-1α）表达水平。结果 T1-T3时HR组和PHC组血清ALT、AST、TNF-α、IL-1β、HIF-1α和eNOS表达水平明显高于S组（P〈0.05）;与HR组比较,T1-T3时PHC组血清ALT、AST、TNF-α和IL-1β表达明显降低（P〈0.05）。T1、T2时HR和HPC组HIF-1α、eNOS表达水平明显高于S组,但HR组明显低于HPC组（P〈0.05）。结论盐酸戊乙奎醚预处理可减轻大鼠HIRI,其机制可能与盐酸戊乙奎醚上调eNOS、HIF-1α表达和抗炎症作用有关。

Objective To investigate the effects of penehyclidine hydrochloride（PHC）preconditioning on hepatic ischemia-reperfusion injury（HIRI）in rats.Methods Fifty-four male SpragueDawley rats,weighing 230-250 g,were randomly divided into three groups（n=18each）：sham operation group（group S）,HIRI group（group HR）and penehyclidine hydrochloride group（group PHC）.In group S,the hepatoduodenal ligaments of rats were only pulled and separated,then abdomens were closed.In group HR,an atraumatic vascular clip was placed on the vessels blocking the portal venous blood supply to the median and left lateral lobes of the liver for 45 minutes,which resulted in approximately 70%rat liver ischemia injury.In group PHC,the rats were treated with 0.45mg/kg penehyclidine hydrochloride at 30 minutes before the portal venous and hepatic arterial were blocked like group HR.Animals were killed at 2h（T1,n=6）,4h（T2,n=6）,24h（T3,n=6）after HIRI or sham surgery.Liver tissues and blood samples were taken for analysis.The serum concentration of ALT and AST were measured as the markers of hepatic functional damage.The TNF-a and IL-1βconcentration were measured by the enzyme linked immunosorbent assay（ELISA）technique.The endothelial nitric oxide synthase（eNOS）and hypoxia-inducible factor（HIF-1α）expression were measured by immunohistochemical staining.Results Compared with group S,the expressions of ALT,AST,TNF-αand IL-1βin groups HR and PHC increased at T1-T3（P〈0.05）,which were similar to the change trend of HIR-1αand eNOS expressions（P〈0.05）.Compared with group HR,the expression of ALT,AST,TNF-αand IL-1βin group PHC decreased at T1-T3（P〈0.05）.Nevertheless expression of HIF-1αand eNOS of groups HR and HPC were increased than that of group S,and experssion of HIF-1αand eNOS of group HPC were increased than that of group HR at T1 and T2（P〈0.05）.Conclusion PHC preconditioning can protect the liver from HIRI.The mechanism may be associated with the up-regulation of eNOS and HIF-1α,as well as reducing the inflammatory response.