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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 role of phosphorylated p38 mitogen-activated protein kinase (p-p38MAPK) in acute liver failure induced by galactosamine (D-Gal N) or lipopolysaccharide (LPS) (ACLF) in patients with liver expression and its significance. Methods C57BL / 6 mice were induced by D-Gal N / LPS to establish acute liver failure model. Experimental groups were established at 0, 0.5, 1, 2, 4, 6, The liver tissue samples were taken for HE staining to observe the pathological changes of hepatic tissue structure. The expression of p-p38MAPK in liver tissues was detected by semi-quantitative Western blotting and immunohistochemical staining respectively. Meanwhile, the expression of pp38MAPK in HBV-ACLF, Hepatitis cirrhosis, Chronic hepatitis B (CHB) in patients with liver tissue expression. Comparison between groups using independent samples t test. Results Western blotting showed that the expression of p-p38MAPK in liver homogenates of mice with liver failure continued to increase over time, and the expression of p-p38MAPK in 6 h group was significantly higher than that of normal control group (P <0.05). The difference was statistically significant t = -2.727, P = 0.034). The results of immunohistochemistry showed that with the prolongation of survival time, the severity of liver inflammation in mice gradually increased. The expression of p-p38MAPK was mainly in sinusocytes in the early stage of inflammation. The expression of p-p38MAPK was gradually increased in liver cells with necrosis of the liver A large number of p-p38MAPK positive hepatocytes were distributed around the tissue. The expression of p-p38MAPK in normal human liver tissue was very low. The expression of pp38MAPK in infiltrating lymphocytes and hepatocytes in the liver tissue of CHB patients showed an increasing tendency with clinical progression Observe the degree of lesion gradually aggravate. Conclusions The expression of p-p38MAPK in D-Gal N / LPS-induced murine model of acute liver failure is increased with the damage of liver tissue, indicating that it plays an important role in the pathogenesis of liver failure. In the pathogenesis of HBV-ACLF, p -p38MAPK signaling pathway may play an important role.