急性肝衰竭大鼠高迁移率族蛋白B1的变化和意义  

Changes in expression of high mobility group protein B1 in acute liver failure in rats

作　　者：张宇[1] 王刚[1] 祝文彩[2] 陈钟[1] 

机构地区：[1]南通大学附属医院肝胆外科,江苏省南通市226000 [2]南通大学附属医院检验科,江苏省南通市226000

出　　处：《世界华人消化杂志》2015年第9期1402-1410,共9页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.81070360;江苏省研究生创新计划基金资助项目;No.CXZZ13_0876;南通大学研究生创新计划基金资助项目;No.ykc13074~~

摘　　要：目的:比较高迁移率族蛋白B1(high mobility group protein B1,HMGB1)与其他炎症因子在急性肝衰竭(acute liver failure,ALF)大鼠肝组织和血清中出现时间及持续时间,探讨其来源及作用.方法:采用腹腔内注射D-氨基半乳糖(D-galactosamine,D-Gal)和脂多糖(lipopolysaccharide,LPS)制备大鼠ALF模型,以腹腔内注射生理盐水作为对照组.于注射后3、6、12、24、48、72 h检测血清中肝功能的变化,观察肝组织病理,实时荧光定量PCR检测肝组织中HMGB1、白介素1β(interleukin 1β,IL-1β)、IL-6、肿瘤坏死因子-α(tumor necrosis factor-α,T N F-α)m R N A变化,E L I S A法检测血清中HMGB1、IL-1β、IL-6、TNF-α水平变化,免疫组织化学检测肝组织中H M G B 1表达变化.使用重组高迁移率族蛋白B1(recombinant high mobility group protein 1,r HMGB1)单独尾静脉注射或联合D-G a l、L P S腹腔内注射,观察大鼠的一般情况并计算生存率.结果:采用D-Gal和LPS成功建立大鼠ALF模型.ALF大鼠肝组织中HMGB1 m RNA表达和血清中HMGB1水平高峰均较IL-1β、I L-6、T N F-α出现晚,但其高峰持续时间更长.D-Gal和LPS注射后的3 h,免疫组织化学可见肝细胞中HMGB1由细胞核转移到细胞质中;注射后24-48 h,肝脏正常组织结构消失,HMGB1蛋白自坏死的肝细胞被动释放.添加外源性r HMGB1使大鼠死亡时间提前,ALF大鼠在相同时间点死亡率升高.结论:大鼠ALF过程中HMGB1由坏死的肝细胞被动释放,其出现时间较其他炎症因子为晚.HMGB1与其他炎症因子相互作用,可能进一步促进ALF中的炎症反应.AIM：To observe the changes in the expression of high mobility group protein B1（HMGBl）and other inflammatory cytokines in acute liver failure（ALF） in rats.METHODS：D-galactosamine（D-Gal） and lipopolysaccharide（LPS） were used to establish a model of ALF by intraperitoneal injection.Rats were treated with normal saline alone in a control group.Serum and liver tissues were collected at different time points（3,6,12,48,72 and 96 h）.Serum biochemical indicators were detected,and HMGBl expression in liver tissue was observed by immunohistochemical analysis.HE staining was performed to evaluate the severity of liver damage.The changes of HMGBl,interleukin1β（IL-1β）,IL-6,and tumor necrosis factor-α（TNF-α） mRNA expression were determined by real-time fluorescent quantitative PCR,and the levels of HMGBl,IL-ip,IL-6 and TNF-a were measured using ELISA kits.rHMGBl was injected via the tail vein alone or combined with D-Gal and LPS by intraperitoneal injection,and the changes of symptoms and signs and survival rates of rats were observed.RESULTS：ALF was successfully induced in rats using D-Gal and LPS.In ALF rats,the peak of HMGBl gene expression and serum levels of HMGBl were later but lasted longer than IL-ip,IL-6 and TNF-a.Cytoplasmic translocation of HMGBl was observed as early as 3 h after D-Gal and LPS administration.In 24-48 h,normal liver tissue structures disappeared,and HMGBl was overflowed from necrotic liver cells and showed diffused yellow staining,full of the entire field of vision.Administration of exogenous rHMGBl reduced the time to death and increased mortality rates.CONCLUSION：HMGB1 may be passively leaked by necrosis hepatic cells,which appeared later compared with other inflammatory cytokines.The interaction of HMGB1 and other inflammatory cytokines can promote the inflammation progression in ALF.

关 键 词：高迁移率族蛋白B1 急性肝衰竭 炎症因子 全身炎症反应综合症 

分 类 号：R575.3[医药卫生—消化系统]