黄芩甙抗肝星状细胞增殖及胶原合成的作用

Inhibitory effects of Baicalin on proliferation of hepatic stellate cells and synthesis of extracellular matrix

目的研究黄芩甙对大鼠肝星状细胞活化、增殖及细胞外基质合成的作用,探讨黄芩甙抗肝纤维化的机制。方法采用胶原酶循环灌流法和密度梯度离心法分离肝星状细胞。不同浓度黄芩甙作用后,通过观察细胞贴壁及细胞形态变化情况来检测细胞活化;采用MTT法检测细胞增殖;免疫细胞化学法检测Ⅰ、Ⅲ型胶原及层粘连蛋白的合成。结果黄芩甙(75～1 200μg/mL)可抑制HSC活化,MTT法示75μg/mL、150μg/mL、300μg/mL、600μg/mL、1 200μg/mL黄芩甙作用于HSC的A值分别为(0.060±6.53)×10-3、(0.052±7.38)×10-3、(0.036±1.26)×10-3、(0.023±1.72)×10-3、(0.013±4.01)×10-3,空白对照组A值为(0.065±1.32)×10-4,F值=1 147.611,P<0.05。黄芩甙可抑制活化的HSC增殖,减少HSCⅠ、Ⅲ型胶原及层粘连蛋白合成。结论黄芩甙抗肝纤维化作用主要机制可能为黄芩甙抑制肝星状细胞活化、增殖,抑制细胞胶原蛋白及糖蛋白等细胞外基质合成。

Objective To study the effects of Baicalin on cells activation, proliferation, collagen and laminin synthesis of cultured rat hepatic stellate cells （HSCs） and to investigate its mechanism of reducing liver fibrosis. Methods Hepatic stellate cells were isolated by in stiu collagenase perfusion and density gradient eentrifugation. HSCs were incubated with various concentrations of Baicalin for 12, 24 and 48 hours. Cell activation was determined by observing cell adherence and morphology. Cells proliferation was measured by MTT colorimetric assay. Collagen and laminin synthesis were detected by immunoeytochemieal staining. Results Baicalin significantly inhibited activation and proliferation of HSCs. The absorbance values of the MTTassay were （0.065 ±1.32） ×10^-4, （0.060±6.53）×10^-3, （0.052 ±7.38） ×10^-3, （0.036±1.26） ×10^-3, （0.023 ±1.72） ×10^ 3, （0.013±4.01） ×10^-3, respectively for the blank control and the concentration 75 μg/mL, 150 μg/mL, 300 μg/mL, 600 μg/mL, 1200 μg/mL of Baicalin （F = 1 147. 611, P 〈0.05）. Moreover, Baicalin decreased the synthesis of type Ⅰ , Ⅲ collagen and laminin synthesis in aetived HSCs, Conclusion Baicalin has a remarkable antifibrotic effect for the liver. The mechanisms are maily due to its effect of inhibiting the activation and proliferation of HSCs, suppressing the synthesis of extraeellular matrix such as collagen and glyeoprotein.