磷酸化p38丝裂原活化蛋白激酶在急性肝衰竭小鼠模型及HBV相关慢加急性肝衰竭患者肝组织中的表达及意义  

Study on p-p38 expression and its significance in liver tissues of mice with acute liver failure and patients with HBV-related acute-on-chronic liver failure

作　　者：陈婧[1] 刘晓燕[1] 杨昊臻[1] 黄坤[1] 胡瑾华[1] 辛绍杰[1] 

机构地区：[1]解放军第三〇二医院肝衰竭诊疗与研究中心,北京100039

出　　处：《临床肝胆病杂志》2015年第4期556-559,共4页Journal of Clinical Hepatology

基　　金：国家自然科学基金面上项目(81171641);首都临床特色应用研究(Z131107002213157)

摘　　要：目的研究磷酸化后的p38丝裂原活化蛋白激酶(p-p38MAPK)在氨基半乳糖(D-Gal N)或脂多糖(LPS)诱导的急性肝衰竭小鼠模型以及HBV相关慢加急性肝衰竭(ACLF)患者肝脏中的表达及其意义。方法采用D-Gal N/LPS诱导C57BL/6小鼠构建急性肝衰竭模型,分别在给药0、0.5、1、2、4、6、8 h设立实验组,每组4只,处死小鼠取肝组织标本进行HE染色观察肝组织结构的病理变化,分别运用蛋白免疫印迹技术半定量检测及免疫组化染色定位检测肝组织中p-p38MAPK的表达;同时对照研究pp38MAPK在HBV-ACLF、乙型肝炎肝硬化、慢性乙型肝炎(CHB)患者肝组织中的表达情况。组间比较采用独立样本t检验。结果蛋白免疫印迹法检测p-p38MAPK在肝衰竭小鼠肝组织匀浆中的表达随时间变化持续增高,给药6 h组半定量分析表达量显著高于正常对照组,差异具有统计学意义(t=-2.727,P=0.034)。免疫组化染色结果显示随存活时间的延长,小鼠肝组织炎症程度逐渐加重,炎症早期主要为窦细胞表达p-p38MAPK,随着肝组织损害加重,肝细胞表达p-p38MAPK渐多,坏死肝组织附近分布大量p-p38MAPK阳性肝细胞;正常人肝组织中p-p38MAPK的表达量很低,CHB患者肝组织内可见浸润淋巴细胞及肝细胞表达pp38MAPK,并且随病程进展呈增多趋势,与临床观察病变程度逐渐加重相一致。结论 D-Gal N/LPS诱导的小鼠急性肝衰竭模型中,p-p38MAPK表达随肝组织损害加重,表明其在肝衰竭病变过程中占重要地位;在HBV-ACLF患者致病过程中,p-p38MAPK信号通路可能发挥重要作用。Objective To investigate the expression and significance of phosphorylated p38mitogen-activated protein kinase （p-p38MAPK） in mice with acute liver failure induced by D-galactosamine （D-GalN）/lipopolysaccharide （LPS）and patients with HBV-related acute-on-chronic liver failure （HBV-ACLF）.Methods C57BL/6 mice induced by D-GalN/LPS were used to establish an acute liver failure model,and experimental groups were set up at 0,0.5,1,2,4,6,and 8 hours （n=4 for each group）.All mice were sacrificed and the samples of liver tissues were given HE staining to observe pathological changes.Western blotting and immunohistochemical staining were used to perform semiquantitative analysis and detect the expression of p -p38MAPK in liver tissues.Meanwhile,the expression of p -p38MAPK in patients with HBV-ACLF,hepatitis B cirrhosis,and chronic hepatitis B was also observed.Independent-samples t test was used to draw comparison between groups.Results Western blotting showed that the expression of p-p38MAPK in liver tissue homogenate increased with time.The semiquantitative analysis showed that the expression was significantly higher in the 6 h group than in the control group （t=-2.727,P=0.034）.Immunohistochemical staining showed that liver inflammation aggravated with the survival time,and p-p38MAPK was expressed by sinus cells at early stage,and then by liver cells with the damage continued in mice,and there were lots of p-p38MAPK-positive liver cells around the necrotic liver tissues.Expression of p-p38MAPK was very low in normal human liver tissues;however, tumor-infiltrating lymphocytes and p-p38MAPK-positive liver cells could be seen in liver tissues of patients with chronic hepatitis B.The expression of p-p38MAPK was increasingly observed with the course of the disease,which was consistent with the aggravation of the dis-ease.Conclusion The expression of p-p38MAPK is found positively correlated with the liver tissue damage in the mouse model of acute liver failure induced by D-GalN/LPS,implying that p-p38

关 键 词：肝炎病毒 乙型 肝功能衰竭 P3 8丝裂原活化蛋白激酶类 疾病模型 动物 

分 类 号：R575.3[医药卫生—消化系统] R-332[医药卫生—内科学]