BMP9通过ALK1/2信号通路诱导肝祖细胞成熟分化的作用研究  

BMP9 Induces Mature Differentiation of Liver Progenitor Cells through ALK1/2 Signaling Pathway

作　　者：刘代江 万晓强 陈佳[1] 李雯[1] 王银光[2] LIU Daijiang;WAN Xiaoqiang;CHEN Jia;LI Wen;WANG Yinguang(Department of Gastroenterology,Chongqing University Central Hospital(Chongqing Emergency Medical Center),Chongqing 400010,China;Department of Proctology,Chongqing Traditional Chinese Medicine Hospital,Chongqing 400021,China)

机构地区：[1]重庆大学附属中心医院(重庆市急救医疗中心)消化内科,重庆400010 [2]重庆市中医院肛肠科,重庆400021

出　　处：《中国细胞生物学学报》2022年第7期1311-1318,共8页Chinese Journal of Cell Biology

基　　金：重庆市渝中区基础研究与前沿探索项目(批准号:20210172)资助的课题。

摘　　要：该研究探讨了骨形态发生蛋白9(BMP9)通过ALK1/2信号通路诱导肝祖细胞成熟分化的作用。通过腺病毒介导siALK1和siALK2感染肝祖细胞14-19(hepatic progenitor cell 14-19,HP14-19),Real-time PCR及Western blot分别检测ALK1及ALK2的表达,荧光素酶报告基因检测ALB-Gluc活性,Real-time PCR检测肝脏相关基因AFP、ALB、CK18、ApoB的mRNA水平表达,PAS染色和ICG摄取实验检测肝细胞的代谢及糖原合成功能。结果显示,腺病毒介导的siALK1和siALK2可特异性抑制HP14-19细胞内ALK1和ALK2的表达,BMP9可诱导HP14-19的成熟分化,细胞形态呈现多角形铺路石样,ALB-Gluc读数显著增加,肝干细胞标志物AFP下调,成熟肝细胞标志物ALB、CK18及ApoB表达显著上调,ICG和PAS染色阳性细胞数增多,Ad-siALK1和Ad-siALK2组,肝细胞标志物表达下降,解毒代谢及糖原合成能力下降。总之,BMP9可通过ALK1/2信号通路诱导肝祖细胞成熟分化。This study aimed to investigate the effect of BMP9(bone morphogenetic protein 9)on induction of mature differentiation of liver progenitor cells through ALK1/ALK2 signaling pathway.HP14-19(hepatic progenitor cell 14-19)was infected with adenovirus-siALK1 and siALK2.The expressions of ALK1 and ALK2 are detected by Real-time PCR and Western blot.Relative acitvity ALB-Gluc was detected by luciferase reporter assay and the mRNA expression of liver related genes AFP,ALB,CK18 and ApoB were detected by Real-time PCR.PAS staining and ICG uptake assay were used to detect the metabolism and glycogen synthesis of hepatocytes.The results showed that Ad-siALK1 and Ad-siALK2 infection could specifically inhibit the expression of ALK1 and ALK2 in HP14-19 cells,and BMP9 could induce the mature differentiation and hepatic function of HP14-19 cells.With BMP9 induction,cell morphology showed polygonal paving stone shape,ALB-Gluc activity was significantly increased,and the expression of liver stem cells marker AFP was down-regulated,while the expressions of hepatic related markers ALB,CK18 and ApoB in mature hepatocytes were significantly up-regulated,and the number of ICG and PAS staining positive cells increased.In AdsiALK1 and Ad-siALK2 treated groups,the expressions of hepatic mature markers,detoxification metabolism and glycogen synthesis decreased.In conclusion,BMP9 might induce mature differentiation of liver progenitor cells through ALK1/ALK2 signaling.

关 键 词：骨形态发生蛋白9 活化素受体样激酶1/活化素受体样激酶2 肝祖细胞 分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]