培养肝内胆管上皮细胞摄取大肠杆菌脂多糖并表达肿瘤坏死因子-α-mRNA

Uptake of Escherichia coli lipopolysaccharide and expression of tumor necrosis factor α mRNA by isolated rat intrahepatic bile duct epithelial cells

目的观察分离培养肝内胆管上皮细胞对大肠杆菌脂多糖的摄取作用及其肿瘤坏死因子－α－ｍＲＮＡ的表达。方法利用免疫荧光、荧光原位杂交及激光共聚焦扫描技术。结果无血清状态下，培养肝内胆管上皮细胞加入Ｓ型脂多糖（Ｓ－ＬＰＳ）培养１５分钟后，其胞浆内抗脂多糖荧光强度显著高于空白对照水平；与Ｓ－ＬＰＳ培养３小时后其胞浆中肿瘤坏死因子－α－ｍＲＮＡ含量增强，６小时达高峰。胞核中亦有肿瘤坏死－α－ｍＲＮＡ增多，但增加幅度低于胞浆。结论结果提示，分离培养的肝内胆管上皮细胞能摄取脂多糖入胞浆，且强烈表达肿瘤坏死因子－α－ｍＲＮＡ。

Objecitve\ To study the uptake of E.coli lipopolysaccharide(LPS)and expression of tumor necrosis factor α mRNA in isolated intrahepatic bile duct epithelial cells.Methods\ Fluorescent immunohistochemical and in situ hybridization techniques and observations with aconfocal laser scanning microscope.Results\ Positive reactions to LPS were found in the cytoplasm of isolated intrahepatic bile duct epithelial cells after incubation with Stype LPS(S LPS)for15minutes,and the FITC fluorescent intensity against LPS was significantly higher than that of controls.After incubation with S LPS for3 hours,FITC fluorescent intensities to the expression of tumor necrosis factor(TNF) α mRNA by flurescent in situ hybridization in the cytoplasm and nuclei of cultured bile duct epithelial cells were sig nificantly higher germinal layers were studied.Tumor like lesions,4 types,112 cases(3.5%);benign tumors,10 types,499 cases(15.8%);malignant tumors,11types, 2 549 cases(80.7%).Of the three groups,the most common being inflammatory pseudotumors,ac counting for 73.2% of tumor like lesions,cavernous hemangioma,accounting for 74.3% of benign tumors and hepatocellular carcinoma,accounting for 96.8% of malignant tumors respectively.The association of cirrhosis in small hepatocellular carcinoma(<3cm)was as high as 83.2%. Conclusion\ A clos relationship between HBV,liver cirrhosis and hepatocellular carcinoma exists.