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目的:通过检测肝龙胶囊联合复方甘草酸苷用药后人肝星状细胞系(LX-2)中I型胶原(Col-I),III型前胶原(PCIII),IV型胶原(IV-C),层粘蛋白(LN),透明质酸(HA),转化生长因子-β1(TGF-β1)的含量及Col-I,基质金属蛋白酶抑制剂1(TIMP-1)mRNA的表达变化,探讨肝龙胶囊联合复方甘草酸苷对LX-2细胞的抗肝纤维化作用。方法:体外培养LX-2细胞,将对数期细胞分为复方甘草酸苷用药组(3.125,1.562,0.781 g·L~(-1)),空白组,肝龙胶囊(0.938,0.469,0.235 g·L~(-1))联合复方甘草酸苷(3.125,1.562,0.781 g·L~(-1))交叉用药组,采用酶联免疫吸附测定(ELISA)法检测肝纤维化四项指标PCIII,IVC,LN,HA和Col-I,TGF-β1的含量;逆转录PCR(RT-PCR)检测Col-I和TIMP-1 mRNA的表达水平。结果:与复方甘草酸苷用药组比较,联合用药组在24,48 h时均可使HA,Col-I的含量降低(P<0.05),在72 h可使LN,HA,Col-I,PCIII,TGF-β1的含量降低(P<0.05);与空白组比较,联合用药组可使TGF-β1含量明显降低(P<0.05,P<0.01);与复方甘草酸苷用药组比较,联合用药组明显降低TIMP-1 mRNA的表达量(P<0.05),且呈现时间依赖性;与空白组比较,联合用药组对Col-I mRNA在24 h呈现抑制作用(P<0.05),在72 h时却使其表达量增加。结论:肝龙胶囊联合复方甘草酸苷用药组可使LX-2细胞LN,HA,Col-I,TGF-β1的含量降低,抑制Col-I和TIMP-1 mRNA的表达,效果优于复方甘草酸苷用药组。肝龙胶囊增强了复方甘草酸苷的体外抗肝纤维化作用。
OBJECTIVE: To detect the expression of type I collagen, type III procollagen (PCIII), type IV collagen (IV-C) in human hepatic stellate cell line (LX-2) ), Laminin (LN), hyaluronic acid (HA), transforming growth factor-β1 (TGF-β1) and the expression of Col-I and TIMP- Anti - hepatic fibrosis effect of liver capsule combined with compound glycyrrhizin on LX-2 cells. Methods: The LX-2 cells were cultured in vitro and the logarithmic phase cells were divided into compound glycyrrhizin group (3.125,1.562,0.781 g · L -1), blank group, liver capsule (0.938,0.469,0.235 g · L -1) combined with compound glycyrrhizin (3.125,1.562,0.781 g · L -1) were used to detect the four indexes of liver fibrosis by enzyme-linked immunosorbent assay (ELISA) PCIII, IVC, LN, HA, Col-1 and TGF-β1 were detected by RT-PCR. The expression of Col-I and TIMP-1 mRNA was detected by RT- Results: Compared with the compound glycyrrhizin group, the content of HA and Col-I in the combination group decreased (P <0.05) at 24 h and 48 h, and the levels of LN, HA, Col-I, (P <0.05, P <0.05). Compared with the blank group, the content of TGF-β1 in the combination group was significantly decreased (P <0.05, P <0.01); Compared with the compound glycyrrhizin group, Compared with the blank group, the combined treatment group inhibited the expression of Col-I mRNA at 24 h (P <0.05), while the expression of TIMP-1 mRNA in the treated group decreased significantly (P <0.05) h, but its expression increased. Conclusion: Combination of compound capsules and glycyrrhizin can reduce the content of LN, HA, Col-I and TGF-β1 in LX-2 cells and inhibit the expression of Col-I and TIMP-1 mRNA, Glucoside medication group. Liver capsule enhances compound glycyrrhizin in vitro anti-liver fibrosis.