| 摘要: |
| 目的：观察不同浓度丁酸钠对大鼠肝卵圆细胞系WB-F344细胞生长、分化的影响，探讨丁酸钠体外诱导WB-F344细胞分化为胆管上皮细胞的条件及分化规律。方法：不同浓度丁酸钠（0．75、2．25、3．75、4．5mmol／L）作用于WB-F344细胞（丁酸钠处理组），观察细胞形态学的变化，MTT法检测细胞生长情况，3．75mmol／L丁酸钠作用WB-F344细胞后，免疫组化观察细胞角蛋白19（CK19）蛋白表达的变化；RT-PCR观察CK19、β4-整合蛋白、γ-谷胺酰转肽酶（GGT）以及甲胎蛋白（AFP）、白蛋白（ALB）mRNA表达的变化。以同期常规培养未作处理的WB-F344细胞作为空白对照组。结果：丁酸钠处理后，各组WBF344细胞生长均受到抑制，3．75、4．5mmol／L丁酸钠处理组光密度值明显低干空白对照组（P〈0．01），而0．75、2．25mmol／L组与空白对照组无明显差异；3．75、4．5retool／L丁酸钠诱导后细胞变大变圆，细胞核增大，核质比减小，而0．75、2．25mmol／L组细胞形态无显著变化。3．75mmol／L丁酸钠处理组WB-F344细胞CK19阳性率为（92．3±1．1）％，明显高于空白对照组（1．3±0．2）％（P〈0．01）。3．75mmol／L丁酸钠处理组可见β4-integrin表达，而空白对照组未见表达；GGT、CK19表达较空白对照组增强；空白对照组见AFP表达，而3．75mmol／L丁酸钠处理组未见表达；ALB在两组均未见表达。结论：3．75mmol／L丁酸钠适合体外诱导大鼠肝卵圆细胞系WB-F344细胞向胆管上皮细胞分化 |
| 关键词: WB-F344细胞 丁酸钠 胆管 上皮细胞 细胞分化 |
| DOI：10.3724/SP.J.1008.2007.00032 |
|
| 基金项目: |
|
| Sodium butyrate induces rat liver oval cells WB-F344 differentiating into billiary epithelium cells in vitro |
| TANG Qing-he, YANG Wen, TAN Ye-xiong, ZHOU Wei-ping |
| (1. Department of Hepatic Surgery Ⅲ, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai 200438, China; 2. International Cooperation Laboratory on Signal Transduction, Eastern Hepatobiliary Surgery Institute) |
| Abstract: |
| Objective：To investigate the effect of sodium butytate （different concentrations） on the growth and proliferation of rat liver oval cell line WB-F344, and to discuss the conditions and rules for sodium butyrate-inducd WB-F344 cells differentiation into biliary lineage in vitro. Methods： WB-F344 cells were treated with sodium butyrate （0.75, 2.25, 3.75, 4.5 mmol/L） and the cell growth and morphological changes were observed; routinely cultured WB-F344 cells were taken as control. The changes of CK19 protein expression were examined immunohistochemically after WB-F244 cells were treated with 3.75% sodium butyrate; and the expression of phenotypic markers, such as 7-glutamyltransferase （GGT） ,β4-integrin, CK19, AFP and ALB at mRNA level were determined by RT-PCR. Untreated WB-F344 cells were used as blank control. Results： We found that sodium butyrate inhibited the growth of WB-F344 cells. The optical densities were significantly decreased in 3.75 and 4. 5 mmol/L groups compared with that in control group（P〈0.01）; but no significant difference was found between 0. 75, 2.25 mmol/L groups with control group. WB-F344 cells treated with 3. 75, 4. 5 mmol/L sodium butyrate became larger and round, with increased nuclei and decreased nucleus to cytoplasm ratio; those treated with 0.75, 2.25 mmol/L had no obvious changes. Immunohistochemical results showed that sodium butyrate significantly increased CK19 expression compared with control group（92.3±1.1]% vs [1. 3±0. 2]% , P〈0.01）. RT-PCR showed increased expression of β4-integrin in sodium butyrate treated groups, but not in control group; the expression of GGT and CK19 was higher than that of control group. Alpha-fetoprotein （AFP） expression was observed in blank control group, but not in sodium butyrate treated cells. Albumin expression was not detected in the 2 groups. Conclusion.. Sodium butyrate at 3. 75 mmol/L is suitable for inducing WB-F344 cells differentiate into the biliary lineage in vitro |
| Key words: WB-F344 cells sodium butyrate bile ducts epithelial cells cell differentiation |
