四氯化碳腹腔注射致大鼠肝纤维化机理探究.docVIP

四氯化碳腹腔注射致大鼠肝纤维化机理探究 　 作者：叶国荣，舒建昌，何雅军，吕霞 【摘要】 目的 观察四氯化碳腹腔注射制作大鼠肝纤维化模型时相关细胞因子的变化，初步探讨四氯化碳腹腔注射致大鼠肝纤维化的机理。方法 以四氯化碳腹腔注射的方法[四氯化碳0.025 mL (用花生油1∶6稀释)/次，每周3次，共14周]制作大鼠肝纤维化模型，设立正常、溶剂对照组，检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、透明质酸(HA)、层黏连蛋白(LN)、Ⅲ型前胶原(PCⅢ)含量;ELISA法检测转化生因子β1(TGFβ1)、肿瘤坏死因子(TNFα);化学法检测血清氧化亚氮(NO)。肝组织行HE和Masson胶原染色，光镜下观察病理学改变，并按SSS计分系统进行评分。免疫组化染色观察肝组织血小板源生长因子BB(PDGFBB)的变化，专业图像分析软件进行图像分析。结果 与正常对照组、溶剂对照组比较，模型组血清ALT、AST、HA、LN、PCⅢ、NO、TGFβ1 、TNFα明显升高(Plt;0.05)，大鼠肝组织肝纤维化评分明显升高(Plt;0.05)，肝组织PDGFBB表达明显增多(Plt;0.05)。结论 促进相关细胞因子的表达可能是四氯化碳腹腔注射致大鼠肝纤维化的机理之一。 【关键词】 四氯化碳;肝纤维化;细胞因子 Abstract:Objective To investigate the variation of correlated cytokine when the rat models of hepatic fibrosis were induced by intraperitoneal injection of carbon tetrachloride,and to the relevant molecular mechanism. Methods Rat models of hepatic fibrosis were made by intraperitoneal injection of carbon tetrachloride at 0.025 mL(diluted 1∶6 with peanut oil),three times a week,a total of 14 weeks. The normal and dissolvent groups were prepared as controls. Serum levels of ALT,AST,HA,LN,PCⅢ were detected; serum levels of TGFβ1 and TNFα were detected by ELISA method. Serum levels of NO were detected by chemical method. HE and Masson staining were conducted in hepatic tissues to observe pathological variations. Grades of hepatic fibrosis were evaluated according to semiquantitative histological scoring system (SSS system). Immunohistochemical staining was executed for detecting PDGFBB in liver,and professional software for image analysis was used. Results Carbon tetrachloride could increase obviously serum levels of ALT,AST,HA,LN,and PCⅢ (Plt;0.05),which were higher than the normaland dissolvent groups. The score of hepatic fibrosis in the model group increased significantly (Plt;0.05). Carbon tetrachloride could increase the expression of PDGFBB (Plt;0.05). Conclusion The correlated cytokines was increased when the rat models of hepatic fibrosis were induced by intraperitoneal injection of car