全反式维甲酸促进骨形态发生蛋白9诱导肝祖细胞成熟分化的作用研究  

The Effect of All-Trans Retinoic Acid on BMP9 Induced Mature Differentiation of Hepatic Progenitor Cells

作　　者：龚梦嘉[1] 惠慧[1] 崔洁洁 毕杨[1] 何昀[1,2] 

机构地区：[1]重庆医科大学附属儿童医院儿科研究所干细胞实验室儿童发育疾病研究教育部重点实验室儿童发育重大疾病国家国际科技合作基地儿科学重庆市重点实验室,重庆400014 [2]重庆医科大学附属儿童医院胃肠新生儿外科,重庆400014

出　　处：《中国细胞生物学学报》2018年第1期17-24,共8页Chinese Journal of Cell Biology

基　　金：重庆市科委基础科学与前沿技术研究项目(批准号:cstc2016jcyjA0228)资助的课题~~

摘　　要：该文研究了全反式维甲酸(all-trans retinoic acid,ATRA)促进骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)诱导肝祖细胞14-19(hepatic progenitor cell 14-19,HP14-19)成熟分化的作用。重组腺病毒Ad-BMP9和ATRA单独及联合作用诱导肝祖细胞HP14-19成熟分化,荧光素酶报告基因检测ALB-Gluc表达情况,Real-time PCR检测肝脏相关基因DLK、AFP、ALB和TAT的mRNA水平,Western blot及免疫荧光检测AFP、ALB、CK18和UGT1A蛋白质水平,PAS染色和ICG摄取实验检测成熟分化后的功能。Ad-BMP9组和ATRA组的ALB-Gluc活性较对照组增高,而AdBMP9+ATRA组ALB-Gluc活性又显著高于Ad-BMP9组和ATRA组。Ad-BMP9+ATRA组的ALB和TAT mRNA水平以及ALB、CK18和UGT1A蛋白质水平均高于Ad-BMP9组和ATRA组,但肝干细胞标志DLK、AFP的表达均降低(P<0.05)。Ad-BMP9+ATRA组的ICG摄取及PAS染色阳性细胞数显著高于Ad-BMP9组和ATRA组。ATRA和Ad-BMP9均诱导肝祖细胞HP14-19成熟分化,联合作用强于单独作用。This study was to investigate the effect of ATRA on Ad-BMP9 induced mature differentiation of hepatic progenitor cells（HP14-19）. Ad-BMP9 and ATRA induced the differentiation of HP14-19, individually or jointly. Luciferase reporter gene was used to detect ALB promoter-driven Gluc activity. Real-time PCR detected the level of mRNA of liver related genes and protein level was detected by Western blot and immunofluorescence. PAS staining and ICG uptake assay were used to detect the function of mature differentiation. Ad-BMP9＋ATRA group could promote the differentiation. The activity of ALB-Gluc, mRNA expression level of ALB and TAT, and protein levels of ALB, CK18 and UGT1 A were higher than that of Ad-BMP9 group and ATRA group. The levels of DLK and AFP were decreased（P〈0.05）, liver function improved significantly. ATRA and Ad-BMP9 induced the mature differentiation of HP14-19, and the combined effect stronger than that of the single effect.

关 键 词：全反式维甲酸 骨形态发生蛋白9 肝祖细胞 分化 

分 类 号：R575.3[医药卫生—消化系统]