肝龙胶囊联合水飞蓟素体外抗肝纤维化的实验研究  

Experimental study of anti-liver fibrosis of Ganlong capsules combined with silymarin in vitro

作　　者：邵明园 焦素月 李燕 赖泳 Ming-yuan Shao;Su-yue Jiao;Yan Li;Yong Lai(College of Chemistry and Pharmacy,Dali University,Dali,Yunnan 671003,Chain;Yunnan Provincial Key Laboratory of Entomological Biopharmaceutical Research and Development,Dali University,Dali,Yunnan 671003,China)

机构地区：[1]大理大学药学与化学学院,云南大理671003 [2]大理大学云南省昆虫生物医药研发重点实验室,云南大理671003

出　　处：《中国现代医学杂志》2018年第29期36-42,共7页China Journal of Modern Medicine

基　　金：云南省教育厅科学研究基金重大专项(No:ZD2014013)

摘　　要：目的观察肝龙胶囊联合水飞蓟素对人肝星状细胞系(LX-2)转化生长因子-β1(TGF-β1)、肝纤维化指标表达的影响,以及体外抗肝纤维化作用。方法体外培养LX-2细胞,将对数期细胞分为水飞蓟素用药组、肝龙胶囊联合水飞蓟素用药组、阴性组。采用酶联免疫吸附法检测TGF-β1、Ⅰ型胶原(Col-Ⅰ)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(Ⅳ-C)、层粘连蛋白(LN)、透明质酸(HA)水平的变化;逆转录聚合酶链反应(RT-PCR)检测Col-I、基质金属蛋白酶抑制剂1(TIMP-1)mRNA的表达。结果与水飞蓟素用药组相比,肝龙胶囊联合水飞蓟素用药组TGF-β1水平降低(P <0.05);24和48 h时,LX-2细胞LN、HA、Col-Ⅰ水平降低(P <0.05);24 h时,PCⅢ、Ⅳ-C水平降低(P <0.05)。与阴性组比较,肝龙胶囊联合水飞蓟素用药组在高浓度联合时可降低LN、HA、Col-Ⅰ、Ⅳ-C的含量(P<0.05),在低浓度时则增加LN、HA、Col-Ⅰ、Ⅳ-C的含量(P <0.05),对PCⅢ、TGF-β1的水平无影响(P>0.05)。与水飞蓟素用药组相比,肝龙胶囊联合水飞蓟素用药组降低TIMP-1 mRNA的表达(P <0.05),在高浓度联合并在72 h时降低Col-ⅠmRNA的表达(P <0.05),在低浓度联合时则增加Col-ⅠmRNA的表达(P <0.05)。结论肝龙胶囊联合水飞蓟素可通过减少细胞外基质的生成,增强水飞蓟素体外抗肝纤维化作用。Objective To observe the influence of Ganlong capsules combined with silymarin on expression of transforming growth factor beta 1(TGF-β1)in human hepatic stellate cells,hepatic fibrosis indexes and the antifibrosis effect in vitro.Methods Human hepatic stellate cell line(LX-2)was cultured in vitro.Enzyme-linked immunosorbent assay was used to detect the levels of TGF-β1,hepatic fibrosis indexes type I collagen(Col-I),type III procollagen(PCIII),type IV collagen(IV-C),laminin(LN)and hyaluronic acid(HA).RT-PCR was used to detect the expressions of Col-I and TIMP-1 mRNAs.Results Compared with the compound silymarin treatment group,the content of TGF-β1 significantly decreased(P<0.05),the content of LN,HA and Col-Ⅰin the LX-2 cells decreased at the 24th and 48th h(P<0.05),and the content of PCIII and IV-C decreased significantly at the 24th h(P<0.05)in the Ganlong capsules combined with silymarin treatment group.Compared with the negative group,Ganlong capsules combined with silymarin treatment in high concentration could reduce the content of LN,HA,Col-I and IV-C(P<0.05),and increased the content of LN,HA,Col-I and IV-C with low concentration(P<0.05),did not have effect on the content of PCIII or TGF-β1(P>0.05).Compared with the silymarin treatment group,Ganlong capsules combined with silymarin treatment significantly decreased the expression of TIMP-1 mRNA(P<0.05),and in high combined concentration significantly decreased the expression of Col-I mRNA at the 72th h(P<0.05),at low combined concentration increased the expression of Col-I mRNA(P<0.05).Conclusions Ganlong capsules can enhance the anti-fibrosis effect of silymarin by reducing the generation of extracellular matrix in vitro.

关 键 词：肝龙胶囊 水飞蓟素 肝纤维化 人肝星状细胞 

分 类 号：R575.2[医药卫生—消化系统]