五味子油联合莪术油防治肝纤维化的作用及机制研究  

Effect and mechanism of Schisandra chinensis oil combined with Zedoary Turmeric oil on liver fibrosis

作　　者：侯晓荣 赵靖 赵佳 丁凯欣 刘文龙[3] 肖小河 湛小燕 柏兆方 HOU Xiao-rong;ZHAO Jing;ZHAO Jia;DING Kai-xin;LIU Wen-long;XIAO Xiao-he;ZHAN Xiao-yan;BAI Zhao-fang(Department of Hepatology,The Fifth Medical Center of Chinese PLA General Hospital,Beijing 100039,China;School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;School of Pharmacy,Hunan University of Traditional Chinese Medicine,Changsha 410208,China;China Military Institute of Chinese Materia,The Fifth Medical Centre of Chinese PLA General Hospital,Beijing 100039,China)

机构地区：[1]中国人民解放军总医院第五医学中心肝病医学部,北京100039 [2]成都中医药大学药学院,四川成都611137 [3]湖南中医药大学药学院,湖南长沙410208 [4]中国人民解放军总医院第五医学中心全军中医药研究所,北京100039

出　　处：《中草药》2022年第4期1059-1067,共9页Chinese Traditional and Herbal Drugs

基　　金：国家“重大新药创制”科技重大专项(2017ZX09301022);国家自然科学基金重点项目(81930110)。

摘　　要：目的研究五味子油联合莪术油防治肝纤维化的作用及机制。方法采用蛋氨酸和胆碱缺乏及铁添加氨基酸饲料喂养小鼠构建非酒精脂肪肝炎伴肝纤维化模型,将112只C57BL/6小鼠分为对照组、模型组、扶正化瘀胶囊(585 mg/kg)组、五味子油单独给药组、莪术油单独给药组以及五味子油联合莪术油给药组;造模成功后,对照组和模型组ig 0.5%羧甲基纤维素钠溶液,各给药组ig相应药物,连续6周;采用酶标仪测定小鼠血清中丙氨酸转氨酶(alanine aminotransferase,ALT)和天冬氨酸转氨酶(aspartate aminotransferase,AST)活性;采用苏木素-伊红(HE)染色法观察各组小鼠肝组织病理变化;采用ELISA法检测小鼠血清中转化生长因子-β1(transforming growth factor-β1,TGF-β1)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和肝组织中羟脯氨酸(hydroxyproline,Hyp)含量;采用qRT-PCR法检测小鼠肝组织中I型胶原(Collagen I)、TGF-β1和Smad3的m RNA表达情况。人肝星形LX-2细胞给予药物预处理,再加入TGF-β1因子,采用Western blotting法检测细胞Collagen I和α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)蛋白表达情况。结果体内实验结果显示,与模型组相比,各给药组血清中ALT和AST活性均显著降低(P<0.05、0.01、0.001),其中五味子油与莪术油联合给药高剂量组抑制小鼠ALT活性更为显著;肝脏颜色、光泽和质地都有明显改善,肝组织中脂肪空泡化改善、炎性细胞浸润减少;肝组织Hyp和血清TGF-β1、TNF-α含量均显著降低(P<0.05、0.01、0.001),其中联合给药高剂量组改善小鼠肝组织Hyp含量更为显著;肝组织Collagen I、TGF-β1和Smad3 mRNA表达水平显著降低(P<0.05、0.01、0.001)。体外实验结果显示,五味子油与莪术油联合给药组能下调TGF-β1诱导的LX-2细胞Collagen I和α-SMA蛋白表达。结论五味子油和莪术油均具有抗肝纤维化作用,且五味子油与莪术油联用的抗肝纤维化效应明�Objective To explore the effect and mechanism of Schisandra chinensis oil combined with Zedoary Turmeric oil on prevention and treatment of liver fibrosis.Methods A mice model of nonalcoholic steatohepatitis(NASH)with liver fibrosis was constructed by methionine and choline deficiency and iron-supplemented amino acid diet.Totally 112 C57BL/6 mice were randomly divided into control group,model group,Fuzheng Huayu Capsules(585 mg/kg)group,S.chinensis oil group,Zedoary Turmeric oil group and S.chinensis oil combined with Zedoary Turmeric oil group.After successful modeling,mice in control and model group were ig 0.5%CMCNa,and other groups were ig corresponding drug for six weeks.The activities of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in serum were analyzed by microplate analyzer.The pathological changes of liver tissue were observed by HE staining.The contents of transforming growth factor-β1(TGF-β1),tumor necrosis factor-α(TNF-α)in serum and hydroxyproline(Hyp)in liver tissue were determined by ELISA.The mRNA expressions of Collagen I,TGF-β1 and Smad3 in liver tissue were determined by qRT-PCR.LX-2 cells were pretreated with drugs,and then TGF-β1 factor was added,protein expressions of Collagen I andα-smooth muscle actin(α-SMA)were detected by Western blotting.Results The results of in vivo experiments showed that compared with model group,ALT and AST activities in serum of mice in each administration group were significantly reduced(P<0.05,0.01,0.001),in which high-dose S.chinensis oil combined with Zedoary Turmeric oil group decreased ALT activity more significantly.The liver color,luster,texture and fat vacuolization of liver tissue in each treatment group were significantly improved,accompanied with the reduction of inflammatory cell infiltration.The contents of Hyp in liver tissue and TGF-β1,TNF-αin serum in each administration groups were decreased(P<0.05,0.01,0.001),among which the combination of high dose group improved Hyp content in liver tissues of mice more significan

关 键 词：五味子油 莪术油 肝纤维化 LX-2细胞 转化生长因子-β/Smad3通路 

分 类 号：R285.5[医药卫生—中药学]