肝素对肝纤维化大鼠肝星状细胞增殖及分泌的影响及其作用机制

Effects of heparin on proliferation and incretion of hepatic stellate cells in fibrotic rats

目的观察肝素在体外对肝纤维化大鼠肝星状细胞增殖及分泌的影响,并探讨其作用机制。方法应用Nycodenz分离肝纤维化大鼠的肝星状细胞置培养板中,并分为三组。A、B组分别加入1000μg/ml及2000μg/ml肝素,C组不用药。应用MTT比色法检测各组肝星状细胞的增殖状况,免疫细胞化学检测各组肝星状细胞中α-平滑肌肌动蛋白(α-SMA)、转化生长因子-β1(TGF-β1)、层粘连蛋白(LN)的表达。结果OD值A组为0.0626±0.0137,B组为0.0746±0.0131,C组为0.1106±0.0198,A、B组均显著低于C组(P均<0.05),B组低于A组(P>0.05);C组肝星状细胞胞质中可见α-SMA、TGF-β1和LN明显表达,A、B组各指标表达均低于C组(P分别<0.01和<0.05),A组表达低于B组(P<0.05)。结论肝素可抑制肝纤维化大鼠肝星状细胞的增殖及胶原分泌,作用机制可能是抑制肝星状细胞表达TGF-β1。

[Objective：] To evaluate the effects of heparin on the proliferation and secretion of hepatic stellate cells （HSCs） ,and explore the mechanism. [Methods] HSCs were isolated by one-step density gradient centrifuge with nycodenz and divided into 3 groups. A and B group were given 1 000 μg/ml and 2 000 μg/ml heparin respectively. HSCs proliferation was detected by MTT colorimetric assay. The expressions of α-smooth muscle actin （α-SMA）, transforming growth factor,beta 1 （TGF-β1） and laminin （LN） were measured with immunocytochemistry. [Results] The OD values of HSCs in A,B and C group were 0. 0626±0. 0137,0. 0746±0. 0131 and 0. 1106±0. 0198,respectively. The OD value of HSCs in A and B group was lower than that in C group significantly （P〈0. 05） ,and the OD value of HSCs in B group was lower than that in A group. The expression of α-SMA,TGF-β1 and LN in HSCs of A and B group was weaker than that in HSCs of group C. [Conclusion] Heparin can inhibit the proliferation of HSCs and the collagen synthesis,it may be related with the inhibition of TGF-β1 expression in HSCs.