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目的 观察乙醇对体外培养枯否细胞(KCs)吞噬功能、微丝表达和膜流动性影响.方法 经门静脉插管胶原酶灌流消化肝脏,密度梯度离心分离KCs;体外培养KCs中加入不同剂量乙醇(1、2、4g/L)作用3、24 h,采用聚苯乙烯乳珠吞噬实验、荧光染色法和荧光光度法检测KCs的吞噬功能、微丝表达、肌动蛋白含量及膜流动性.结果 1、2、4g/L乙醇组作用3、24 h KCs吞噬聚苯乙烯乳珠数量分别为(16.05 ±3.69)、(15.86±4.50)、(13.86 ±4.69)和(13.95±4.51)、(14.35±6.52)、(9.71±3.34)个,明显低于对照组[分别为(20.13 ±5.26)、(20.30 ±8.57)个](P＜0.05);4g/L乙醇组(作用24 h)KCs吞噬功能明显低于1、2 g/L乙醇组(P＜0.05);各剂量乙醇组(作用3h)KCs微丝排列紊乱,但肌动蛋白含量无明显变化;与对照组(5.85±1.13)比较,2、4g/L乙醇组KCs膜流动性[(2.30±1.08)、(1.62±0.30)]明显降低(P＜0.05),呈剂量依赖性.结论 乙醇可降低体外KCs的吞噬功能,其机制可能与乙醇致KCs膜流动性下降和微丝排列紊乱有关.“,”Objective To observe effects of ethanol on phagocytosis,actin expression,and membrane fluidity in Kupffer cells (KCs) in vitro.Methods KCs were isolated from rats with collagenase perfusion and density ladder centrifugadon technique.The isolated KCs were cultured with ethanol (1,2,and 4 g/L) for 3 or 24 hours;then,phagocytosis,actin expression,and membrane fluidity of the KCs were determined with beads phagocytosis test,fluorescence staining,and fluorescence polarization.Results The number of phagocytosed beads for the KCs treated with ethanol at 1,2,and4 g/L were 16.05 ±3.69,15.86 ±4.50,and 13.86 ±4.69 at 3-hour and 13.95 ±4.51,14.35 ±6.52,and 9.71 ± 3.34 at 24-hour,which were significantly lower than those of the control group (20.13 _± 5.26 and 20.3 ± 8.57;P ＜ 0.05 for all);the phagocytic function of the KCs treated with 4 g/L ethanol for 24 hours was significantly lower than that of KCs treated with 1 and 2 g/L ethanol (both P ＜ 0.05).The microfilaments of KCs in all ethanol treatment groups were arranged in a disorderly manner,but there was no significant difference in actin expression among the groups (P ＞ 0.05).Compared with that of the control group (5.85 ± 1.13),the membrane fluidity of KCs in 2 and 4 g/L ethanol treatment groups (2.30 ± 1.08 and 1.62 ± 0.30) was decreased in a dose-dependant manner significantly (both P ＜ 0.05).Conclusion Ethanol could decrease the phagocytosis of KCs in vitro,and the effect may be related to the changes in membrane fluidity and the expression of microfilaments of in KCs.