一氧化氮在大鼠梗阻性黄疸继发肝功能损伤中的作用  

Effect of nitric oxide on hepatic dysfuction in rat obstructive jaundice

作　　者：涂巍[1] 智迎辉[1] 郭仁宣[1] 

机构地区：[1]中国医科大学附属第一医院普外二科,辽宁省沈阳市110001

出　　处：《世界华人消化杂志》2004年第4期875-879,共5页World Chinese Journal of Digestology

基　　金：中国医科大学博士后启动基金项目;No.2002-157~~

摘　　要：目的:探讨一氧化氮(NO)在梗阻性黄疸合并肝功能障碍中的作用. 方法:将56只Wistar大鼠随机分成假手术对照组和胆总管结扎组(实验组).应用胶原酶灌注法分离肝细胞并进行培养; 用白介素-1β(IL-1β)等细胞因子处理培养的肝细胞,应用Griess法和Western印迹分析法检测两组NO及诱导型一氧化氮合酶(iNOS)的产生;应用高效液相色谱法(HPLC)检测两组肝细胞5’三磷酸腺苷(ATP)的合成;应用酶法检测两组肝细胞酮体含量并计算酮体比率(乙酰乙酸盐/β-羟基丁酸盐) (KBR). 结果:实验组NO产生量明显高于对照组(207.99μmoL/L vs 78.57 μmoL/L P<0.01);NO的产生与IL-1β作用时间和作用剂量有关;实验组和对照组的iNOS产生量在蛋白水平没有差异;IL-1β降低了实验组培养肝细胞的ATP含量(15.94 nmoL/106 cells vs 20.21 nmoL/106cells,P<0.05), 对照组改变不明显;IL-1β降低了两组的KBR,实验组降低程度大于对照组;一氧化氮合酶(NOS)抑制剂NG-甲基- L-精氨酸(L-NMMA)抑制了NO的产生,使降低的肝细胞ATP含量、KBR得以恢复. 结论:梗阻性黄疸能够促进肝细胞产生NO,导致肝功能障碍.L-NMMA可以缓解由于大量NO导致的ATP合成障碍及KBR下降.对NO产生的调节可能会成为预防及治疗梗阻性黄疸继发肝功能损害的有效方法.AIM: To examine whether obstructive jaundice influences the production of nitric oxide and alters hepatic functions including energy metabolism. METHODS: Hepatocytes were isolated by perfusing the liver with collagenase and cultured from a rat model of obstructive jaundice or sham-control. Rat cultured hepa-tocytes were incubated with cytokines including IL-1β. The production of nitric oxide was measured with Griess reagent method and Western blot analysis. Adenine nucleotides (ATP, ADP and AMP) were measured by a high performance liqiud chromatography. Ketone bodies in the medium were measured enzymatically. RESULTS: Obstructive jaundice increased the production of nitric oxide by IL-1β time-and dose- dependently. Western blot analysis revealed that protein levels of iNOS were unchanged between two groups. IL-lp decreased the ATP content and KBR in obstructive jaundice but not that in sham-control. Addition of L-NMMA blocked the decreases of ATP content and KBR as well as nitric oxide production. CONCLUSION: Enhancement of nitric oxide production following obstructive jaundice is associated with the alteration of hepatic energy metabolism through mitochondrial dysfuntion, resulting in liver failure. Regulation of nitric oxide may be a useful therapy for preventing liver damage in obstructive jaundice.

关 键 词：一氧化氮 NO 大鼠 梗阻性黄疸 肝功能损伤 肝功能衰竭 

分 类 号：R575[医药卫生—消化系统] R-332[医药卫生—内科学]