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目的探讨脂多糖(LPS)对肝脏Kupffer细胞增殖和分泌功能及超微结构的影响。方法将小鼠原代Kupffer细胞培养扩增后,随机分为LPS组和正常对照组。2组细胞培养24 h后,LPS组加入LPS,继续培养6 h,甲基噻唑基四唑法(MTT)测定细胞增殖情况,流式细胞仪检测细胞周期变化,透射电镜观察细胞超微结构;并收集细胞上清液,检测肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、白介素-6(IL-6)水平。结果 LPS组吸光度值高于正常对照组,G0/G1期细胞分布低于正常对照组,S+G2/M期细胞分布高于正常对照组(P<0.01)。LPS组细胞上清液中TNF-α、IL-1β和IL-6水平高于正常对照组(P<0.01)。LPS组Kupffer细胞内可见大量空泡及自噬体形成,正常对照组个别细胞胞质内仅见少量空泡。结论 LPS能激活肝脏Kupffer细胞的增殖和分泌功能,并可引发肝脏Kupffer细胞自噬和超微结构改变。
Objective To investigate the effects of lipopolysaccharide (LPS) on the proliferation, secretion and ultrastructure of hepatic Kupffer cells. Methods Primary murine Kupffer cells were cultured and expanded, then randomly divided into LPS group and normal control group. After 24 h of culture, LPS group was added with LPS for 6 h. MTT assay was used to detect cell proliferation. Cell cycle was analyzed by flow cytometry. Cell ultrastructure was observed by transmission electron microscopy. The cell supernatants were collected and the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL- Results The absorbance of LPS group was higher than that of normal control group. The cell distribution in G0 / G1 phase was lower than that in normal control group. The cell distribution in S + G2 / M phase was higher than that in control group (P <0.01). The levels of TNF-α, IL-1β and IL-6 in the supernatant of LPS group were higher than those in the normal control group (P <0.01). A large number of vacuoles and autophagosomes were found in Kupffer cells in LPS group, while only a few vacuoles were found in the cytoplasm of normal cells in normal control group. Conclusion LPS can activate the proliferation and secretion of hepatic Kupffer cells and induce autophagy and ultrastructural changes in liver Kupffer cells.