培养肝细胞,肝窦壁内皮细胞及贮脂细胞对大肠杆菌脂多糖的摄入作用

目的和方法：应用免疫荧光及激光共聚焦扫描技术，观察原代培养大鼠肝细胞、肝脏贮脂细胞及窦壁内皮细胞对不同分子结构大肠杆菌脂多糖的摄取作用。结果：无血清条件下，肝细胞、贮脂细胞、窦壁内皮细胞加入Ｓ型脂多糖（Ｓ－ＬＰＳ）及Ｒｄ型脂多糖（Ｒｄ－ＬＰＳ）培养５ｍｉｎ后，胞浆内均出现强烈抗脂多糖荧光显色；胞浆抗脂多糖荧光强度显著高于空白对照水平；且Ｒｄ－ＬＰＳ培育后肝细胞、窦壁内皮细胞胞浆内荧光强度增高幅度显著高于Ｓ－ＬＰＳ培养后的增高幅度。抗脂多糖显色局限于胞浆，胞核内未见明显着色，但经与Ｒ型多糖培养后，肝细胞核内亦出现强烈抗脂多糖荧光显色。结论：分离培养的大鼠肝细胞、贮脂细胞及肝窦壁内皮细胞对细菌脂多糖具有摄入作用，且脂多糖与这些细胞的相互作用依脂多糖分子结构不同而异

Uptake of Escherichia coli lipopolysaccharides by isolated rat hepatocytes, sinusoidal endothelial and fat storing cells

AIM and METHODS:By using a monoclonal antibody specific to core lipid A region of lipopolysaccharides (LPS) and immunofluorescent technique, the uptake process of different types of LPS by isolated hepatocytes, sinusoidal endothelial and fat storing cells were observed with a cofocal laser scanning microscope. RESULTS:Fluorescent intensities of LPS in the cytoplasm of isolated hepatocytes, fat storing cells and sinusoidal endothelial cells increased significantly after 5 min incubation with wild form LPS(S-LPS) or rough form LPS(Rd-LPS) in serum-free medium. The increase of fluorescent intensity of LPS in the cytoplasm of hepatocytes and sinusoidal endothelial cells after incubation with S-LPS was much higher than that of Rd-LPS. Positive fluorescent reactions of LPS were only flund in the cytoplasm of the cells. However fluorescent reactions against LPS were also found in nuclei of hepatocytes after incubation with R-LPS. CONCLUSION:Isolated hepatocytes, sinusoidal endothelial and fat storing cells could take up LPS into the cytoplasm, and that interactions of LPS with hepatocytes might be different according to different chemical structure of LPS.