激活素A对肝星状细胞细胞外基质合成的影响

Effect of Activin on extracelluar matrix secretion in isolated rat hepatic stellate cell

目的:探讨激活A(activin A,ACTA)对肝星状细胞(hepatic stellate cell,HSC)细胞外基质(extracellular matrix,ECM)合成的影响。方法:采用原位酶灌注法和密度梯度离心法分离HSC,初次传代后,HSC随机分为8组:空白对照组(A组)、ACTA1μg/L组(B组)、ACTA10μg/L组(C组)、ACTA100μg/L组(D组)、TGFβ_1 10μg/L组(E组)、ACTA1μg/L+TGFβ_1 10μg/L组(F组)、ACTA 10μg/L+TGF β_1 10μg/L组(G组)、ACTA 100μg/L+TGF β_1 10μg/L组(H组),加药后24h,采用放射免疫法检测培养液Ⅲ型前胶原、Ⅳ型胶原含量及半定量RT-PCR方法测定细胞Ⅲ型胶原mRNA表达。结果:ACTA能刺激体外培养HSC分泌ECM,在一定浓度范围内(1-100μg/L)呈剂量依赖关系;100μg/L ACTA刺激HSC分泌ECM的能力与10μg/L TGF-β_1相当,而且ACTA能协同TGF-β_1发挥这一生物学效应。结论:激活素A参与肝纤维化形成。

AIM: To investigate the effect of activin A on the extracelluar matrix secretion of rat hepatic stellate cell. METHODS: Hepatic stellate cels were isolated and purified from normal male Sprague-Dawley rat liver by a combination of pronase-collagenase perfusion and density gradient centrifugation. Passaged hepatic stellate cells were divided randomly into eight groups: control group(A group), ACTA1 μg/L group (B group), ACTA 10 μg/L group(C group), ACTA 100 μg/L group (D group), TGF β_1 10 μg/L group(E group), TGF β_1 10 μg/L plus ACTA 1 μg/L group(F group), TGF β, 10 μg/L plus ACTA 10 μg/L group(G group), TGF β_1 10 μg/L plus ACTA 100 μg/L group(H group). 24 h after incubation secretion of procollagen Ⅲ, collagen Ⅳ and mRNA of collagen Ⅲ in hepatic stellate cells were detected by radioimmunoassays and semi-quantitative RT-PCR method respectively. RESULTS: Extracellular matrix secretion in passaged hepatic stellate cells was enhanced by activin A according to its concentration, the capacity of extracellular matrix secretion by 100 μg/L activin A was equal to that of 10 μg/L TGF β_1, extracellular matrix secretion and type Ⅲ collagen mRNA expression in passaged hepatic stellate cells was enhanced by activin A and TGFβ_1 in a synergistic manner. CONCLUSION: Activin A may contribute to hepatic flbrogenesis.