抗纤软肝颗粒对肝纤维化大鼠Hedgehog通路核转录因子Gli1的影响  

Effect of Kangxian Ruangan Granule on the Nuclear Transcription Factor Gli1 of Hedgehog Pathway in Rats with Hepatic Fibrosis

作　　者：张爽 江诗怡 高翔[2] 詹磊[2] 陆定波[2] ZHANG Shuang;JIANG Shiyi;GAO Xiang;ZHAN Lei;LU Dingbo(the First Clinical College,Hubei University of Chinese Medicine,Wuhan 430061,China;Department of Hepatology,Hubei Provincial Hospital of Traditional Chinese Medicine,Wuhan 430074,China)

机构地区：[1]湖北中医药大学第一临床学院,武汉430061 [2]湖北省中医院肝病科,武汉430074

出　　处：《中西医结合研究》2023年第1期24-29,共6页Research of Integrated Traditional Chinese and Western Medicine

基　　金：湖北省卫生厅中医药、中西医结合科研一般重点项目(No.2012Z-Y01)。

摘　　要：目的观察抗纤软肝颗粒对肝纤维化模型大鼠Hedgehog通路核转录因子Gli1的影响。方法将60只SPF级Wistar大鼠随机分成空白对照组,模型组,抗纤软肝颗粒低、中、高剂量组和环靶明对照组。除空白对照组外,所有实验大鼠均以四氯化碳(CCl4)复合因素法诱导肝纤维化模型8周。第9周开始给药干预,抗纤软肝颗粒低、中、高剂量组分别给予1.125 g/kg、2.5 g/kg、5 g/kg相应药物灌胃,环靶明对照组给10 mg/kg环靶明灌胃;均连续灌胃2周,灌胃期间造模继续。采用酶联免疫吸附试验检测各组大鼠肝功能指标及Ⅰ型胶原水平;采用Gomori银染法观察各组肝脏组织病理学改变;采用Real-time PCR法检测各组大鼠肝组织Gli1 mRNA表达水平,Western-blot法检测各组大鼠肝组织Gli1蛋白质表达水平,免疫组化法检测各组大鼠肝组织Gli1表达。结果Gomori银染显示,模型组大鼠肝组织中胶原纤维广泛增生,纤维间隔相互连接,假小叶形成;抗纤软肝颗粒各剂量组大鼠肝脏组织纤维化程度较模型组减轻,但不如环靶明对照组。与模型组比较,抗纤软肝颗粒中、高剂量组、环靶明组血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天冬氨酸氨基转移酶(aspartate aminotransferase,AST)水平显著降低(P<0.05),且高剂量组显著低于低剂量组(P<0.05);与模型组比较,抗纤软肝颗粒低、中、高剂量组Ⅰ型胶原含量明显降低(P<0.01),且高剂量组明显低于低、中剂量组(P<0.05);与环靶明对照组比较,抗纤软肝颗粒高剂量组ALT、AST水平及I型胶原含量差异无统计学意义(P>0.05)。与模型组比较,抗纤软肝颗粒各剂量组Gli1 mRNA及蛋白表达均降低(P<0.01);与环靶明对照组比较,抗纤软肝颗粒中、高剂量组Gli1 mRNA差异无统计学意义(P>0.05),而抗纤软肝颗粒各剂量组Gli1蛋白表达均增加(P<0.05)。免疫组化结果显示,抗纤软肝颗粒各剂量组大鼠肝脏组织Gli1的表达较模型Objective To observe the effect of Kangxian Ruangan granule on the nuclear transcription factor Gli1 of Hedgehog pathway in rats with hepatic fibrosis.Methods Sixty SPF Wistar rats were randomly divided into blank control group,model group,low,middle and high dose groups of Kangxian Ruangan granule and cyclopamine control group.Except the blank control group,all experimental rats were induced by carbon tetrachloride(CCl4)compound factor method for 8 weeks.At the 9th week,the drug administration intervention was started.The low,middle and high dose groups of Kangxian Ruangan granule were given corresponding drugs of 1.125 g/kg,2.5 g/kg and 5 g/kg respectively by gavage,and the cyclopamine group was given 10 mg/kg of cyclopamine by gavage.The rats were gavaged continuously for 2 weeks,and the modeling continued during gavage.The liver function indexes and the level of typeⅠcollagen were detected by enzyme-linked immunosorbent assay.Gomori silver staining was used to observe the histopathological changes of liver in each group.Real-time PCR method was used to detect the expression level of Gli1 mRNA in liver tissue of rats in each group.Western-blot method and immunohistochemistry were used to detect the expression of Gli1 protein in liver tissue of rats in each group.Results Gomori silver staining showed that the collagen fibers in the liver tissues of rats in the model group were extensively proliferated.Besides,fiber septa were interconnected,and pseudolobules were formed.The degree of fibrosis in the liver histology of rats in each dose group of Kangxian Ruangan granule was less than that in the model group,but not as good as that in the cyclopamine control group.Compared with the model group,the levels of serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in the middle and high dose groups of Kangxian Ruangan granule and the cyclopamine group were significantly decreased(P<0.05),and the high dose group was significantly lower than the low dose group(P<0.05).Compared with the model group,the

关 键 词：肝纤维化 抗纤软肝颗粒 HEDGEHOG通路 GLI1 

分 类 号：R575.2[医药卫生—消化系统] R285.5[医药卫生—内科学]