摘要
目的探讨骨形态发生蛋白9(BMP-9)在胆道闭锁(BA)肝纤维化中的作用机制。方法选取14例BA患儿肝组织标本为BA组,5例胆总管囊肿(CBD)患儿肝组织标本为CBD组,HE染色对肝组织进行肝纤维化评估,免疫组化染色检测BMP-9、p-SMAD1/5表达情况,实时荧光定量逆转录聚合酶链反应(qPCR)检测肝组织BMP-9及DNA结合抑制因子1(ID1)mRNA表达水平。培养人肝星状细胞LX-2,用重组转化生长因子(rTGF)-β1与重组BMP(r BMP)-9处理,蛋白质印迹法检测细胞中SMAD1/5、p-SMAD1/5、ID1蛋白及α-平滑肌肌动蛋白(α-SMA)表达情况,qPCR检测细胞中α-SMA及ID1 mRNA表达情况。结果BA组肝组织中BMP-9及p-SMAD1/5蛋白、BMP-9及ID1mRNA表达水平均高于CBD组;在BA组中重度肝纤维化患儿BMP-9蛋白、BMP-9及ID1 m RNA的表达高于轻度肝纤维化患儿(P<0.05)。LX-2细胞经rTGF-β1、rBMP-9处理后,α-SMA蛋白及α-SMA m RNA表达水平均升高(P<0.05)。LX-2细胞加入不同剂量rBMP-9之后,其下游通路中p-SMAD1/5、SMAD1/5、ID1、α-SMA蛋白,及ID1、α-SMAmRNA表达均较0μg/L组升高(P<0.05)。结论在BA患儿肝组织中BMP-9、p-SMAD1/5、ID1表达水平较CBD患儿增高,与肝纤维化严重程度有关。BMP-9可通过SMAD/ID1信号通路激活人肝星状细胞LX-2,促进纤维化进程。
Objective To explore the mechanism of BMP-9 in biliary atresia(BA)liver fibrosis.Methods Fourteen samples of BA cases were selected as the BA group and 5 samples of congenital biliary dilatation(CBD)cases were selected as the CBD group.Liver fibrosis was evaluated by HE staining,immunohistochemical staining was used to detect the expression levels of BMP-9 and p-SMAD1/5,and qPCR"was used to detect the expression levels of BMP-9 and ID1 mRNA in liver.Human hepatic stellate cells(LX-2)were cultured and treated with rTGF-β1 and different concentrations of rBMP-9.The protein expression levels ofα-SMA,SMAD1/5,p-SMAD1/5 and ID1 in cells were detected by Western blot assay,and the mRNA expressions ofα-SMA and ID1 in cells were detected by qPCR.Results The expression levels of BMP-9 and p-SMAD1/5 proteins,BMP-9 and ID1 mRNA in liver were higher in the BA group than those of the CBD group.In the BA group,the expressions levels of BMP-9 protein,BMP-9 and ID1 mRNA were significantly higher in children with moderate and severe liver fibrosis than those in children with mild liver fibrosis(P<0.05).After treatment with rTGF-β1 and rBMP-9,the expression levels ofα-SMA protein andα-SMA mRNA were increased in LX-2 cells(P<0.05).The expressions of p-SMAD1/5,SMAD1/5,ID1,α-SMA protein and ID1,α-SMA mRNA in the downstream pathway-were increased after adding different concentrations of rBMP-9 in LX-2 cells compared with those of 0μg/L group(P<0.05).Conclusion The expression levels of BMP-9,p-SMAD1/5 and ID1 in the liver of BA children are increased than those of CBD children,which are positively correlated with the degree of liver fibrosis.BMP-9 can activate LX-2 cells through SMAD/ID1 signaling pathway and promote the progress of fibrosis.
作者
葛亮
苟庆云
赵金凤
张聪
陈灵芝
胡晓丽
詹江华
GE Liang;GOU Qing-yun;ZHAO Jin-feng;ZHANG Cong;CHEN Ling-zhi;HU Xiao-li;ZHAN Jiang-hua(Department of General Surgery,Tianjin Children's Hospital,Tianjin 300134,China;Graduate School,Tianjin Medical University;Department of Pathology,Tianjin Children's Hospital)
出处
《天津医药》
CAS
北大核心
2021年第10期1020-1025,共6页
Tianjin Medical Journal
基金
新疆维吾尔自治区自然科学基金资助项目(2019D01A12)
天津市研究生科研创新项目资助(2019YJSS192)。
作者简介
葛亮(1986),男,博士,住院医师,主要从事小儿肝胆疾病方面研究。E-mail:geliangjiayou@163.com;通信作者:詹江华,E-mail:zhanjianghuatj@163.com。
