【摘要】 目的　探讨内毒素 (LPS)及其刺激的Kupffer细胞对大鼠肝星状细胞 (HSC)增殖的影响。方法　用链酶蛋白酶和胶原酶原位灌流 ,Nycodenz密度梯度离心分离大鼠肝脏HSC和Kupffer细胞 ,制备LPS刺激的Kupffer细胞培养上清液 (KCCM ) ,并以MTT法观察LPS及KCCM对HSC增殖的效应。结果　无LPS刺激和浓度为 1μg/ml的LPS刺激所得的KCCM对HSC有显著增殖作用 (P <0 0 1) ,且两组之间有差异 (P <0 0 5 ) ;浓度为 10 μg/ml的LPS刺激所得的KCCM对HSC无影响 (P >0 0 5 ) ;加入兔抗人转化生长因子 β1(TGF β1)可抑制低浓度LPS刺激的KCCM对HSC的增殖作用 (P <0 0 1) ;LPS浓度为 1μg/ml和 10 μg/ml对HSC直接作用均无增殖效应 (P >0 0 5 )。 结论　内毒素刺激的KCCM可促进HSC增殖 ,与肝纤维化的发生可能有关更多还原

【Abstract】 Objective To study the effects of Kupffer cell conditioned medium(KCCM)derived from lipopolysaccharides(LPS)treatment on proliferation of rat hepatic stellate cells(HSC).Methods HSC and Kuppffer cells were isolated from liver of Wistar rats by in situ perfusion with pronase and collagenase and density gradient centrifugation with Nycodenz and then cultured.KCCM was prepared and MTT colorimetric assay was used to detect HSC proliferation.Results HSC and Kupffer cells were isolated with high purity.KCCM derived from LPS (0μg/ml or 1μg/ml)treatment might significantly promote HSC proliferation( P< 0 01),and there was difference between them( P< 0 05); KCCM derived from LPS(10μg/ml)treatment couldn′t promote HSC proliferation( P> 0 05);Adding anti TGF β 1 might suppress the proliferation which was promoted by PLS(1μg/ml)treated KCCM ( P< 0 01); LPS(1μg/ml or 10μg/ml)couldn′t promote HSC proliferation directly( P> 0 05). Conclusion The technique for isolation of HSC and Kupffer cells described here is simple and reliable. KCCM stimulated by LPS may promote HSC proliferation and be associated with hepatic fibrogenesis.更多还原