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

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（PC Ⅲ）、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，PC Ⅲ、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PC Ⅲ、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- Ⅰ mRNA 的表达（P <0.05），在低浓度联合时则增加Col- Ⅰ mRNA 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。

Abstract:

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-Ⅰ in 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.