【摘要】 目的探讨诱生型一氧化氮合酶(iNOS)在阻塞性黄疸肝损害中的作用及果糖的防治机制。方法采用胶原酶原位肝灌注法获取大鼠肝细胞,行原代培养,用iNOS抑制剂SMT作用于肝细胞,再用50μmol/L甘氨鹅脱氧胆酸钠(glycochenodeoxycholate,GCDC)作用后行FCM及TUNEL检测肝细胞凋亡情况;使用不同浓度果糖(Fructose)作用于肝细胞,100μM的GCDC作用后行FCM及TUNEL检测;结扎大鼠胆总管,有和无果糖喂养后3d、7d、14d及21d处死大鼠,分别用TUNEL技术及SABC法检测阻塞性黄疸大鼠肝脏组织细胞凋亡状态及iNOS蛋白的表达。结果随SMT浓度的增加,肝细胞的凋亡明显增加。经果糖作用后,100μM的GCDC致肝细胞的凋亡率明显减少,且随果糖作用浓度的增加肝细胞凋亡率减少;大鼠胆总管结扎后,无果糖作用时,随结扎时间的延长细胞凋亡指数(AI)增加,结扎14d后AI达高峰,iNOS蛋白表达越强,AI就越高。有果糖作用时,iNOS蛋白在14d、21d表达较无果糖作用时明显减少,凋亡指数较无果糖作用时均明显减少。结论果糖通过抑制iNOS的表达在阻塞性黄疸肝损伤中起保护作用。iNOS在阻塞性黄疸肝损害的发生和发展中起重要作用。更多还原

【Abstract】 [Objective] To explore the roles of iNOS in hepatic injury with obstructive jaundice and the cytoprotecting mechanism of Fructose.[Methods] Rat hepatocytes were isolated by in situ collagenase perfusion and primary culture.(1)Heptocytes were pretreated with various concentrations of iNOS inhibitor SMT for 20 min.After pretretment,50 μmol/L GCDC was added for an additional 24 h.Cells were next detected by FCM and TUNEL.(2)After hepatocytes were added with different concentration of Fructose and 100 μM GCDC,cells were evaluated by FCM and TUNEL.(3)Experimental obstructive jaundice(OJ)was induced by double ligation of the bile duct.OJ for 3,7,14,21 days with Fructose and without Fructose.We detected apoptotic status in livers of alle rats with TUNEL and iNOS protein in liver of OJ with immunohistochemistry method.[Results](1)SMT increased GCDC-induced apoptosis in a concentration-dependent manner.(2)Adding with different concentration of Fructose and 100 μM GCDC,the apoptotic rate was decreased and was related to concentration with Fructose.(3)The apoptotic rate of liver is related to times of OJ.Expression of iNOS and apoptosis index(AI)were highest from 14-day bile duct ligation without Fructose.AI and expression of iNOS were decreasing from 14-day bile duct ligation with Fructose.[Conclusions] Fructose is cytoprotective during bile salt-induced apoptosis of hepatocytes by regulating iNOS.iNOS takes part in the regulation and the occurrence and progression of hepatic injury in OJ.更多还原