核因子κB亚基65及细胞色素C氧化酶-Ⅱ在大鼠急性肝衰竭模型中的变化及意义  

Variations and significance of nuclear factor-κB p65 and cytochrome C oxidase-Ⅱ in acute liver failure rat models

作　　者：周莉[1] 陈立艳[2] 马英骥[3] 段钟平[1] 

机构地区：[1]首都医科大学附属北京佑安医院人工肝中心,100069 [2]哈尔滨医科大学附属第二医院感染二科,150000 [3]哈尔滨医科大学附属第四医院感染科,150000

出　　处：《传染病信息》2011年第3期136-139,155,共5页Infectious Disease Information

基　　金：国家重点基础研究发展计划(973计划)专项(2007CB5-12801);国家"十一五"科技重大专项(2008ZX100002-005-03);北京市丰台区科技项目(xm101210)

摘　　要：目的观察核因子κB亚基65(nuclear factor-κB,NF-κB p65)及细胞色素C氧化酶-Ⅱ(cytochrom eCoxidase-Ⅱ,COX-Ⅱ)在大鼠急性肝衰竭(acute liver failure,ALF)模型中的变化。方法 Sprague-Dawley雄性大鼠40只随机分为5组:对照组、ALF4h组、ALF8h组、ALF12h组和ALF24h组。ALF组采用D-氨基半乳糖(800mg/kg)和脂多糖(100g/kg)联合腹腔注射构建ALF模型,对照组注射同等体积的0.9%氯化钠溶液。分别检测各组的ALT、AST、TBIL、ALB、PT、PTA及观察各时间点肝脏病理变化;实时荧光定量PCR法检测各组大鼠肝脏内NF-κBp65的mRNA水平,分光光度法检测各组大鼠肝脏线粒体中的COX-Ⅱ活性。结果光镜结果显示,ALF8h组可见明显肝细胞凋亡,而12h组和24h组可见明显肝细胞坏死。ALF组中8h、12h及24h肝脏NF-κBp65的mRNA水平明显低于对照组(P均<0.001),24h最低。COX-Ⅱ的活性在ALF4h开始升高,8h达到高峰(与对照组比较,P=0.008),12h开始下降,24h下降更为明显。结论 ALF大鼠肝脏NF-κBp65的mRNA水平降低,肝脏线粒体COX-Ⅱ活性在凋亡阶段升高,后期降低。二者均参与了ALF中肝细胞凋亡的发生,同时NF-κBp65还可能抑制了ALF过程中肝细胞的再生。Objective To observe the variations of nuclear factor-kB p65 （NF-kB p65） and cytochrome C oxidase- Ⅱ （COX- Ⅱ） in rat models of acute liver failure （ALF）. Methods Forty male Sprague-Dawley rats were divided into 5 groups： control group, ALF 4-hour group, ALF 8-hour group, ALF 12-hour group, and ALF 24-hour group. All the ALF rats were given 800 mg/kg of D- galactosamine （D-GalN） and 100 g/kg of lipopolysaccharide （LPS） via intraperitoneal （i.p.） injection to establish the ALF model, and the rats in the control group were given 1 ml of 0.9% saline via i.p. injection. Serum alanine aminotransferase, aspartate aminotrans- ferase, total bilirubin, albumin, prothrombin time, prothrombin activity were examined and rat liver pathological changes were observed in each group. Hepatic NF-kB p65 mRNA levels were examined by real time fluorescent quantitative PCR method. Hepatic mitochondrial COX-H activities were examined by speetrophotometry. Results Light microscopic examination showed obvious hepatocyte apoptosis in ALF 8-hour group, and obvious hepatocyte necrosis in ALF 12-hour and 24-hour groups. Hepatic NF-KB p65 mRNA levels in ALF 8-hour, ALF 12-hour and ALF 24-hour groups were lower than those in the control group （P〈0.001）, and the levels in ALF 24-hour group were the lowest. Activities of COX- Ⅱ in ALF rats began to increase at 4 hour, reached a peak at 8 hour （P=0.008, compared with the control group）, began to decrease at 12 hour, and decreased more obviously at 24 hour. Conclusions In ALF rats, hepatic NF-KB p65 mRNA levels decrease, while hepatic mitochondrial COX-II activities increase in apoptosis phase and decrease later. Both NF-KB p65 and COX- Ⅱ participate in hepatocyte apoptosis in ALF, while NF-KB p65 possibly inhibits hepatocyte regeneration.

关 键 词：急性肝衰竭 核因子κB亚基65 细胞色素C氧化酶-Ⅱ 大鼠 

分 类 号：R575.3[医药卫生—消化系统]