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WB-F344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于空白对照组(P<0.01),而0.75、2.25mmol/L组与空白对照组无明显差异;3.75、4.5mmol/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。
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.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2007年第1期32-35,共4页
Academic Journal of Second Military Medical University
