花生四烯酸刺激Kupffer细胞对肝星状细胞增殖的影响

Effect of Kupffer Cells Stimulated by Arachidonic Acid on Proliferation of Hepatic Stellate Cells in Rats

目的探讨花生四烯酸及其刺激的Kupffer细胞（KC）对大鼠肝星状细胞（HSC）增殖的影响。方法用链霉蛋白酶和胶原酶原位灌流，Nycodenz密度梯度离心分离大鼠HSC和KC，并进行鉴定;制备花生四烯酸及其刺激的KC培养的上清液（KCCM），用MTT法观察花生四烯酸及其刺激的KCCM对HSC增殖效应。结果25μg／ml花生四烯酸对HSC增殖有促进作用（P＜0.01），50～100μg／ml花生四烯酸对HSC增殖有抑制作用，显示细胞毒性作用；与正常和KCCM组相比，KCCM+花生四烯酸组对HSC增殖有促进作用（P＜0．01）。结论花生四烯酸在一定浓度下可促进HSC增殖，高浓度对HSC有毒性作用，花生四烯酸及其刺激的KCCM可促进HSC增殖，其可能通过脂质过氧化作用而与脂肪肝肝纤维化的发生有关。

Objective To study the effect of arachidonic acid and kupffer cell- conditioned medium(KCCM) derived from arachidonic acid on proIiferation of hepatic stellatecells(HSC) in rats. Methods HSCs were isolated and cultured from the liver of Wistar ratsby in situ per fusion with pronase and collagenase, as well as by density gradientcentrifugation with Nycodenz. HSC and KC were identificated by immunohistochemistry,endocytosis, and ultrastructure, ect; KCCM were prepared, and HSC proliferation wasdetected by MTT colorimentric assay. Res'Its Arachidonic acid had an effect on prol-iferati0n of HSC; 25pg/ml of arachidonic acid promoted HSC proliferation(P<0.01),but 5O,100pg/ml arachidonic acid had an inhibitory effect, and showed cytotoxity on HSC under theinverted microscope; Compared with the control and KCCM, KCCM+arachidonic acid mightpromote HSC proliferation (P<0.01). KCCM was greater in HSC proliferation than control,but there was no significance statistically (P>0.O5). ConcIusion Arachidonic acid maypromote HSC proliferation, but increased concentration can have cytotoxity. Ara-chidonic acid and KCCM stimulated by it may promote HSC proliferation and be ass0ciatedwith hepatic fibrogenesis by lipid peroxidation.