肿瘤坏死因子α单克隆抗体治疗大鼠肝肺综合征的分子机制  

Molecular mechanism of tumor necrosis factor-α monoclonal antibody in hepatopulmonary syndrome in rats

作　　者：黄学[1] 刘丽[2] 刘楠[3] 

机构地区：[1] 广西贵港市人民医院消化内科 [2] 河北医科大学第二医院消化内科,石家庄050000 [3] 石家庄市第一人民医院

出　　处：《中华肝脏病杂志》2015年第6期458-463,共6页Chinese Journal of Hepatology

基　　金：河北省医学科学研究重点资助课题（08080）

摘　　要：目的 探讨肿瘤坏死因子α单克隆抗体（TNFαMcAb）治疗大鼠肝肺综合征的分子机制. 方法 采用胆总管结扎（CBDL）致大鼠肝肺综合征模型.将60只雄性SD大鼠随机分为假手术组、模型组和干预组.假手术组：仅分离胆总管不结扎（6只大鼠）;模型组：分别于结扎术后1、2、3、4周和5周末麻醉动物取材（每个时间点6只大鼠）;干预组：于结扎术后第2周始于腹腔每2d注射TNF oα McAb 0.1 g/kg,干预后1、2、3周和4周末麻醉动物取材（每个时间点6只大鼠）;假手术组与5周模型组同批麻醉,留取各组动物的肝组织、肺组织.采用HE和Masson染色观察肝组织病理变化;采用HE染色观察肺组织病理变化;采用免疫组织化学方法检测黏着斑激酶（FAK）和磷酸化（p）-FAK在肺组织中的表达定位与表达程度;采用Western blot方法检测FAK、p-FAK和10号染色体同源丢失性磷酸酶与张力蛋白（PTEN）在肺组织中的蛋白表达情况.多组间的均数比较采用单因素方差分析,两组间的比较采用SNK-q检验（方差齐）或Games-Howell检验（方差不齐）. 结果 TNF αMcAb干预组大鼠肝组织的炎症和纤维化程度较模型组明显减轻;干预组大鼠肺组织炎症反应较模型组轻.模型组大鼠肺组织中FAK和p-FAK蛋白含量在术后第1周均高于假手术组,差异具有统计学意义（P值均＜ 0.05）;随着手术时间延长,其蛋白表达量整体呈升高趋势,于术后第3周达到高峰值,各手术时间点与假手术组比较的差异均具有统计学意义（P值均＜ 0.05）.TNFα McAb干预组与假手术组比较,FAK和p-FAK蛋白表达量均高于假手术组,但是低于同手术时间点的模型组,其峰值也出现在术后第3周,免疫组织化学方法得出在干预治疗第1、2、3周时干预治疗组FAK和p-FAK蛋白表达较模型组均降低,差异具有统计学意义（P值均＜0.05）;4周时干预组p-FAK的蛋白含量（0.035±0.005）明显低于模Objective To investigate the molecular mechanism of tumor necrosis factor-alpha （TNF-α） monoclonal antibody （McAb） in hepatopulmonary syndrome using a rat model.Methods Sixty adult male Sprague-Dawley rats,weighing 250±25 g,were randomized to a sham operation group,a common bile duct ligation （CBDL） group,or a CBDL＋TNF-αt McAb treatment group.The CBDL operation group was further divided into five subgroups,and the CBDL＋TNF-α McAb treatment group was further divided into four subgroups.After the experimental period,all rats were sacrificed for excision of lung and liver tissues.Hematoxylin-eosin （HE） and Masson staining were performed to observe the extent of liver fibrosis,and HE staining was used to histopathologically assess changes in the lung tissue.Immunohistochemistry and western blotting were used to investigate the changes in expression levels of FAK,p-FAK and PTEN in lung.Results The extent of inflammatory responses and fibrosis in the liver was significantly lower in the CBDL＋TNF-α McAb treatment group as compared to those in the CBDL group.The inflammatory responses in the lung were also significantly lower in the CBDL＋TNF-α McAb treatment group as compared to that in the CBDL group.The CBDL＋TNF-α McAb treatment group also showed less extensive distribution of FAK and p-FAK protein in lung tissues,but more extensive distribution of PTEN protein.Conclusion FAK and PTEN are associated with hepatopulmonary syndrome in rats.The therapeutic effect of TNF-α McAb may involve modulation of the expression of FAK and PTEN.

关 键 词：肿瘤坏死因子 肝肺综合征 局部黏着斑激酶 10号染色体同源丢失性磷酸酶与张力蛋白 

分 类 号：R392[医药卫生—免疫学]