结缔组织生长因子诱导大鼠前体细胞(WB-F344)向肝脏实质细胞分化  

Differentiation of rat hepatic progenitor cells(WB-F344 cells) into hepatic parenchymal cells induced by connective tissue growth factor

作　　者：杨爱婷[1] 王萍[1] 刘天会[1] 丛敏[1] 白艳锋[1] 丛瑞[1] 吴鹏[1] 尤红[1] 

机构地区：[1]首都医科大学附属北京友谊医院肝病中心,100050

出　　处：《传染病信息》2010年第3期144-147,158,共5页Infectious Disease Information

基　　金：国家自然科学基金(30972602);北京市卫生系统高层次卫生技术人才项目(2009-3-06)

摘　　要：目的观察结缔组织生长因子(connective tissue growth factor,CTGF)对大鼠肝脏前体细胞(WB-F344)分化的影响,进一步观察细胞外基质的变化。方法采用MTT法检测CTGF对WB-F344细胞活性的影响。采用Western blotting方法分别检测CTGF诱导WB-F344细胞后胆管细胞标志物CK-19及肝细胞标记物AFP、ALB的变化。采用Western blotting和实时荧光定量PCR检测细胞外基质相关指标金属蛋白酶组织抑制剂(tissue inhibitor of metallo proteinase,TIMP)-1在mRNA水平和蛋白水平的变化。结果作用于WB-F344细胞24h后,CTGF组的细胞活性大于转化生长因子-β组(P<0.05)。5ng/ml CTGF作用于WB-F344细胞24h后,可以明显改变WB-F344细胞表面标记物在蛋白水平的表达。幼稚肝细胞标记物AFP的表达是对照组的0.38倍(P=0.002),成熟肝细胞标记物ALB和胆管细胞标记物CK-19的表达均增加,分别为对照组的3.5倍和1.58倍(P分别为0.000、0.002)。同时,5ng/ml CTGF可以上调WB-F344细胞TIMP-1在mRNA水平和蛋白水平的表达,分别为对照组的2.47倍和3.36倍(P=0.000、0.002)。结论 5ng/ml CTGF有诱导肝脏前体细胞向肝脏实质细胞分化的趋势,并伴有WB-F344细胞外基质相关指标TIMP-1表达的上调。Objective To observe the influence of connective tissue growth factor （CTGF） on the differentiation of rat hepatic progenitor cells （WB-F344 cells）, and the changes of extracellular matrix during this procedure. Methods MTT assay was used to evaluate the influence of CTGF on the viability of WB-F344 cells. Western blotting was used to detect the expressions of cytokeratin 19 （CK- 19 ）, α-fetoprotein （AFP） and albumin （ALB） after WB-F344 cells were induced by CTGF. Western blotting and real-time fluorescent quantitative PCR were used to detect mRNA and protein expressions of tissue inhibitor of metalloproteinase （TIMP）-1. Results Activity of WB-F344 cells stimulated by CTGF for 24 hours was higher than that stimulated by transforming growth factor-β （P〈0.05）. After WB-F344 cells had been stimulated by CTGF at the concentration of 5 ng/ml for 24 hours, the protein expressions of the phenotype markers of WB-F344 cells changed significantly. The expression of AFP, the fetal hepatocyte marker, was 3.8 times that of the control group （P=0.002）, while the expressions of ALB, the mature hepatocyte marker, and CK- 19, the cholangiocyte marker, 3.5 and 1.58 times those of the control group （P=0.000, 0.002）. Meanwhile, the expressions of TIMP-1 both in mRNA level and protein level were up-regulated when WB-F344 cells were stimulated by CTGF at the concentration of 5 ng/ml, 2.47 and 3.36 times compared with the control group, respectively （P=0.000, 0.002）. Conclusions CTGF at the concentration of 5 ng/ml can induce the differentiation of WB-F344 cells into hepatic parenchymal cells, while the expression of TIMP-1 is found to be up-regulated.

关 键 词：结缔组织生长因子 转化生长因子Β 肝细胞 免疫组织化学 

分 类 号：R333.4[医药卫生—人体生理学] R364[医药卫生—基础医学]