摘要
[目的]探讨经验方二陈汤与苓桂术甘汤对高脂饲料诱导大鼠非酒精性脂肪性肝炎(non-alcoholic steatohepatitis,NASH)分子作用机制。[方法]以高脂饲料喂养40只雄性SD大鼠8周,随机分为正常组、模型组、罗格列酮组、二陈汤组、苓桂术甘汤组,于第9周起分别给予罗格列酮剂量1mg·kg-1、二陈汤4.52g·kg-1、苓桂术甘汤组3.31g·kg-1治疗4周。12周后将大鼠处死并观察药物对大鼠肝功能的影响,并对肝组织进行常规苏木精-伊红染色,观察病理改变;Western blot检测二陈汤与苓桂术甘汤以及两方含药血清对NASH大鼠肝组织以及经软脂酸刺激肝细胞的肿瘤坏死因子TNF-α与核转录因子NF-κB蛋白表达的影响。MTT法检测不同浓度软脂酸对大鼠肝细胞增殖的影响,以及二陈汤与苓桂术甘汤含药血清对软脂酸的拮抗作用。[结果]与模型组比较,罗格列酮和苓桂术甘汤可有效降低NASH大鼠血清ALT、AST(P<0.05),同时改善肝脂肪变性的严重程度和降低TNF-α和NF-κB蛋白的表达(P<0.05)。与模型组比较,虽然二陈汤治疗后NASH大鼠肝组织TNF-α和NF-κB蛋白表达显著性下降(P<0.05),但是血清ALT、AST并未显著性降低(P>0.05),苓桂术甘汤含药血清能明显改善软脂酸对肝细胞增殖的抑制(P<0.05)。与模型组比较,二陈汤与苓桂术甘汤含药血清均能抑制软脂酸刺激的肝细胞TNF-α和NF-κB蛋白表达(P<0.05)。[结论]罗格列酮、二陈汤、苓桂术甘汤能改善大鼠肝脏脂肪变性,降低TNF-α和NF-κB蛋白表达。软脂酸可以明显抑制肝细胞增殖率,罗格列酮与苓桂术甘汤含药血清能拮抗软脂酸对肝细胞增殖的抑制。苓桂术甘汤通过抑制NF-κB蛋白表达,减少TNF-α表达,从而减轻肝脏炎性损伤,进而达到治疗NASH的作用。
[Objective]To observe the effects and mechanism of Erchen and Lingguizhugan decoctione on non-alcoholic steatohepatitis（NASH）rats induced by high fat diet.[Methods]Forty male rats were randomly divided into five groups：the Blank control group,Model group,Rosiglitazone group,Erchen decoctione group,and Lingguizhugan decoctione group.Rats of all groups except the blank control group were fed a high-fat diet for eight weeks to induce NASH.The Rosiglitazone group,Erchen decoctione group,Lingguizhugan decoctione group were respectively treated with Rosiglitazone（1mg·kg-1）,Erchen decoctione（4.52g·kg-1）and Lingguizhugan decoctione（3.31g·kg-1）for four weeks.All rats were killed at 12 weeks and assessed for hepatic histopathological changes by HE.Western blot was used to detect TNF-αand NF-κB protein expression in NASH rat liver and hepatic cells stimulated by palmitic acid.MTT assay was used to detect proliferation of rat hepatocytes stimulated by different concentrations of palmitic acid,and serum containing Erchen and Lingguizhugan decoction＇s antagonism towards palmitic acid.[Results]Compared with the model group,Rosiglitazone and Lingguizhugan decoctione decreased serum ALT and AST levels in NASH rats（P〈0.05）,and also reduced the severity of hepatic steatosis and protein expression of TNF-α/and NF-κB（P〈0.05）.Compared with the model group,although Erchen decoctione decreased protein expression of TNF-αand NF-κB（P〈0.05）,it didn＇t decrease the level of ALT and AST in serum（P〈0.05）.The serum containing Lingguizhugan decoctione improved the hepatocyte proliferation rate that was significantly injured by palmitic acid（P〈0.05）.Compared with the model group,the serum containing Erchen and Lingguizhugan decoction inhibited protein expression of TNF-αand NF-κB of hepatocyte stimulated by palmitic acid（P〈0.05）.[Conclusion]Rosiglitazone,Erchen decoctione and Lingguizhugan decoctione all could reduce the liver steatosis of NASH rats,and also could decrease protein expression of TNF-αand NF-κB.Palmitic acid could obviously inhibit hepatocyte proliferation rate,and the serum containing Rosiglitazone and Lingguizhugan decoctione could protect hepatocyte proliferation injured by palmitic acid in vitro.Lingguizhugan decoction could reduce inflammatory lesions through inhibiting the expression of NF-κB proteins and then decreasing TNF-αexpression.
出处
《中国中西医结合消化杂志》
CAS
2015年第8期525-530,共6页
Chinese Journal of Integrated Traditional and Western Medicine on Digestion
关键词
二陈汤
苓桂术甘汤
非酒精性脂肪性肝炎
大鼠
肿瘤坏死因子-Α
Erchen decoctione
Lingguizhugan decoctione
non-alcoholic steatohepatitis
rat
tumor necrosis factor-a
作者简介
张会存，男，博士，助理研究员，主要从事中医药防治肝病的研究
通讯作者：李军祥，E—mail：Iijx970508@sohu．com
