大鼠肝纤维化肝组织中抗衰老标志蛋白和抗增殖蛋白的表达研究  

Gene expression changes of regucalcin and prohibitin in cirrhotic rat liver and the related effects of compound glutathione inosine injection intervention

作　　者：刘洋[1] 吴君[2] 梁跃东 陆爽[4] 程明亮[4] 刘俊英[1] 

机构地区：[1]周口市中心医院消化科一病区,河南466000 [2]贵阳医学院附属白云医院 [3]贵阳市第五人民医院 [4]贵阳医学院附属医院

出　　处：《中华肝脏病杂志》2014年第11期826-830,共5页Chinese Journal of Hepatology

基　　金：中国肝炎防治基金会王宝恩肝纤维化研究基金（20100034）

摘　　要：目的 研究猪血清所致肝纤维化大鼠肝组织中抗衰老标志蛋白（RGN）、抗增殖蛋白（PHB）表达及谷胱甘肽复方注射液（CGII）对其的干预. 方法 健康雄性Wistar大鼠40只,随机分为正常组、造模组.正常组予等渗盐水腹腔注射0.5 ml/只,造模组予腹腔注射无菌猪血清0.5ml/只,每周两次,连续8周,随机处死造模大鼠4只行病理学检查,验证造模成功后,停止注射猪血清,剩余造模组大鼠随机分为纤维化组和CGII干预组.CGII干预组大鼠给予浓度60 mg/mlCGII肌肉注射;正常组和纤维化组均予等单位体质量容积的等渗盐水肌肉注射,每日一次,连续6周.取大鼠肝组织行HE及Masson染色.RT-PCR和免疫组织化学检测肝组织中RGN、PHB mRNA及蛋白质表达. 计量资料采用单因素方差分析,两两比较采用LDS检验,病理学半定量结果采用秩和检验分析. 结果 纤维化组大鼠肝组织RGN、PHB mRNA相对表达量分别为75.99±12.8、64.54±12.35较正常组的182.09±17.84、192.20±17.12明显减低,F值分别为105.646、347.232,P值均＜0.001,差异均有统计学意义;RGN、PHB相对蛋白表达量分别为10.85±2.81、14.5±2.75较正常组的61.08±7.10、55.86±4.23表达均明显降低,F值分别为226.343、525.889,P值均＜ 0.001,差异均有统计学意义.CGII干预后大鼠肝纤维化程度较纤维化组明显减轻（P值均＜ 0.001）,肝组织RGN、PHB的mRNA和蛋白表达均较纤维化组明显增高,差异有统计学意义（P值均＜ 0.001）.结论 RGN及PHB在猪血清所致大鼠肝纤维化肝组织表达降低.Objective To study expression of regucalcin （RGN） and prohibitin （PHB） genes in cirrhotic rat liver and to investigate the related effects of compound glutathione inosine injection （CGII） intervention.Methods Forty male Wistar rats were randomly divided into a control group （n=12） and a model group （n=28）.The model was established by injecting sterile porcine serum （0.5 mL） into the rat abdominal cavity,twice weekly for 8 consecutive weeks; the control group rats were treated with physiological saline injection （0.5 mL） into the abdominal cavity with the same frequency and time span.During the modeling period,four rats from the model group were randomly selected at different time points to examine changes in liver pathology.Upon pathology confirmation of liver cirrhosis,the porcine serum injection was terminated.The remaining 24 rats in the model group were randomly divided into a fibrosis group and a CGII treatment group.The CGII group received CGII （intramuscular injection of 0.018 mL 100g-1 body weight） once a day for 6 continuous weeks; the fibrosis rats were treated with the same dosage of physiological saline with the same frequency and time span.Liver tissue morphology was examined by both hematoxylin-eosin and Masson＆#39;s staining.RGN and PHB expression at the mRNA and protein levels in liver tissues were detected by real time RT-PCR and immunohistochemical staining,respectively.Results Both the mRNA and protein expression levels of RGN and PHB were significantly lower in the liver tissues of the fibrosis group than in the control group.CGII intervention led to significant alleviation of the liver fibrosis severity; moreover,the mRNA and protein expression levels of RGN and PHB were significantly higher than those in the fibrosis group.Conclusion Down-regulation of regucalcin and prohibitin gene expression might contribute to the pathogenesis of liver cirrhosis.

关 键 词：肝硬化 谷胱甘肽 抗衰老标志蛋白 抗增殖蛋白 

分 类 号：R575.2[医药卫生—消化系统]