[1]艾麦提·,牙森,金鑫,等.TGF-β1信号介导的肝星状细胞促进胚胎肝前体细胞向胆管细胞方向分化[J].第三军医大学学报,2018,40(09):746-752.
 AI MAITI·,Ya Sen,JIN Xin,et al.TGF-β1 signaling mediated hepatic stellate cells promote hepatic progenitor cells differentiation into cholangiocytes [J].J Third Mil Med Univ,2018,40(09):746-752.
点击复制

TGF-β1信号介导的肝星状细胞促进胚胎肝前体细胞向胆管细胞方向分化(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第09期
页码:
746-752
栏目:
基础医学
出版日期:
2018-05-15

文章信息/Info

Title:
TGF-β1 signaling mediated hepatic stellate cells promote hepatic progenitor cells differentiation into cholangiocytes
 
作者:
艾麦提·牙森金鑫陈梓昕王伟李德卫
重庆医科大学附属第一医院肝胆外科,实验研究中心
Author(s):
AI MAITI·Ya Sen JIN Xin CHEN Zixin WANG Wei LI Dewei

Department of Hepatobiliary Surgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China

关键词:
胚胎肝前体细胞肝星状细胞TGF-&beta1共培养
Keywords:
hepatic progenitor cells hepatic stellate cells TGF-&beta1 co-culture
分类号:
R322.47; R329-33; R329.26
文献标志码:
A
摘要:

目的     探讨转化生长因子β1 (transforming growth factor β1, TGF-β1)激活的小鼠肝星状细胞(mouse hepatic stellate cells, mHSCs)对小鼠胚胎肝前体细胞(mouse hepatic progenitor cells, mHPCs)分化的影响。方法    mHSCs细胞株加入10 ng/mL TGF-β1刺激48 h,并分为激活组(mHSCs-TGF-β1)和对照组(mHSCs)。qRT-PCR和Western blot法检测mHSCs激活情况。采用荧光激活细胞筛选法(fluorescence-activated cell sorter, FACS)分选DLK1(delta-like1 homologue)表面抗原阳性的原代mHPCs, 分选的细胞利用免疫荧光染色法观察甲胎蛋白(alpha-fetoprotein, AFP)、白蛋白(albumin, ALB)及细胞角蛋白19(cytokeratin 19, CK19)抗原表达情况。mHPCs与mHSCs Transwell共培养6 d后, 分为mHPCs+mHSCs-TGF-β1组、mHPCs+mHSCs组和mHPCs组。细胞免疫荧光技术法和qRT-PCR法检测分化标志物表达情况。结果    激活组mHSCs中TGF-β1、α-SMA、TGF-βR1、Jagged1、VEGF、HGF等mRNA和TGF-β1、a-SMA、TGF-β-R1、Jagged1等蛋白的表达量均较对照组明显增加(P<0.01);FACS新分选的mHPCs胞质中表达AFP和少量ALB,但不表达CK19;mHPCs+mHSCs-TGFβ1组mHPCs胞质中大量表达胆管细胞标志物,并且CK19、SOX9、Hes1等mRNA表达量均较其他两组明显增加,而其他两组mHPCs中ALB、AFP等的表达量明显增加(P<0.01)。结论    TGF-β1激活的肝星状细胞诱导胚胎肝前体细胞向胆管细胞分化。

Abstract:

Objective    To investigate the effects of mouse hepatic stellate cells (mHSCs) activated by transcription factor β1 (TGF-β1) on mouse hepatic progenitor cells (mHPCs) differentiation. Methods    After the mHSCs were treated with 10 ng/mL TGF-β1 for 48 h, the cells were assigned into activation group (mHSCs-TGF-β1), and those without treatment served as control group (mHSCs). Activation of mHSCs was detected by qRTPCR and Western blotting. mHPCs were isolated from fetal livers based on the cell surface antigen deltalike protein 1 (Dlk1) by a fluorescence-activated cell sorter (FACS), and the expression of alpha-fetoprotein (AFP), albumin (ALB), and cytokeratin 19 (CK19) was observed by immunocytochemical assay. Then the obtained mHPCs and mHSCs were co-cultured in a Transwell system for 6 d. Immunocytochemical assay and qRTPCR were used to detect the expression of differentiation markers in mHPCs+mHSCs-TGF-β1 group, mHPCs+mHSCs group and mHPCs group. Results    The mRNA levels of a-SMA, TGF-β1, TGF-β-R1, Jagged1, HGF and VEGF, and protein levels of TGF-β1, a-SMA, TGF-β-R1 and Jagged1 were significantly higher in the activation group (P<0.01); Freshly sorted mHPCs by FACS expressed AFP and low level of ALB but not CK19 in the cytoplasm. In the mHPCs+mHSCs-TGF-β1 group, mHPCs highly expressed ductspecific markers in the cytoplasm, and the mRNA levels of CK19, SOX9 and Hes1 were obviously higher than the other 2 groups of cells.  But the ALB and AFP expression was more obvious in the 2 groups (P<0.01). Conclusion    TGF-β1-activated hepatic stellate cells induce hepatic progenitor cells differentiation into cholangiocytes.

参考文献/References:

[1]KAMIYA A, INAGAKI Y. Stem and progenitor cell systems in liver development and regeneration[J]. Hepatol Res, 2015, 45(1): 29-37. DOI:10.1111/hepr.12349.
[2]SANCHO-BRU P, NAJIMI M, CARUSO M, et al. Stem and progenitor cells for liver repopulation: can we standardise the process from bench to bedside[J]. Gut, 2009, 58(4): 594-603. DOI:10.1136/gut.2008.171116.
[3]PINTILIE D G, SHUPE T D, OH S H, et al. Hepatic stellate cells’ involvement in progenitormediated liver regeneration[J]. Lab Invest, 2010, 90(8): 1199-1208. DOI:10.1038/labinvest.2010.88.
[4]GORDILLO M, EVANS T, GOUON-EVANS V. Orchestrating liver development[J]. Development, 2015, 142(12): 2094-2108. DOI:10.1242/dev.114215.
[5]SUZUKI K, TANAKA M, WATANABE N, et al. p75 neurotrophin receptor is a marker for precursors of stellate cells and portal fibroblasts in mouse fetal liver[J]. Gastroenterol, 2008, 135(1): 270-281.e3. DOI:10.1053/j.gastro.2008.03.075.
[6]TANIMIZU N, SAITO H, MOSTOV K, et al. Long-term culture of hepatic progenitors derived from mouse Dlk+ hepatoblasts[J]. J Cell Sci, 2004, 117(Pt 26): 6425-6434. DOI:10.1242/jcs.01572.
[7]WEIS S M, CHERESH D A. Tumor angiogenesis: molecular pathways and therapeutic targets[J]. Nature Med, 2011, 17(11): 1359-1370. DOI:10.1038/nm.2537.
[8]FURUYAMA K, KAWAGUCHI Y, AKIYAMA H, et al. Continuous cell supply from a Sox9-expressing progenitor zone in adult liver, exocrine pancreas and intestine[J]. Nat Genet, 2010, 43(1): 34-41. DOI:10.1038/ng.722.
[9]HABIBULLAH C M, SYED I H, QAMAR A, et al. Human fetal hepatocyte transplantation in patients with fulminant hepatic failure[J]. Transplantation, 1994, 58(8): 951-952. DOI:10.1097/00007890-199410270-00016.
[10]ISHII T, YASUCHIKA K, MACHIMOTO T, et al. Transplantation of embryonic stem cell-derived endodermal cells into mice with induced lethal liver damage[J]. Stem Cells, 2007, 25(12): 3252-3260. DOI:10.1634/stemcells.20070199.
[11]KALINICHENKO V V, BHATTACHARYYA D, ZHOU Y, et al. Foxf1 +/- mice exhibit defective stellate cell activation and abnormal liver regeneration following CCl4 injury[J]. Hepatology, 2003, 37(1): 107-117. DOI:10.1053/jhep.2003.50005.
[12]LORENZINI S, BIRD T G, BOULTER L, et al. Characterisation of a stereotypical cellular and extracellular adult liver progenitor cell niche in rodents and diseased human liver[J]. Gut, 2010, 59(5): 645-654. DOI:10.1136/gut.2009.182345.
[13]BAI S, KOPAN R, ZOU W, et al. NOTCH1 regulates osteoclastogenesis directly in osteoclast precursors and indirectly via osteoblast lineage cells[J]. J Biol Chem, 2008, 283(10): 6509-6518. DOI:10.1074/jbc.M707000200.
[14]HU K F, KONG X Y, ZHONG M C, et al. Brucine inhibits bone metastasis of breast cancer cells by suppressing Jagged1/Notch1 signaling pathways[J]. Chin J Integr Med, 2017, 23(2): 110-116. DOI:10.1007/s1165501626472.
[15]肖斌, 陈川宁, 陶忠桦, 等. Notch通路对乙醛激活大鼠肝星状细胞株的增殖及转分化的影响[J]. 中国老年学杂志, 2014, 34(3): 710-713.
XIAO B, CHEN C N, TAO Z H, et al. The role of notch pathway in hepatic stellate cell strain [J]. Chin J Gerontol, 2014, 34(3):710-713.
[16]ANTONIOU A, RAYNAUD P, CORDI S, et al. Intrahepatic bile ducts develop according to a new mode of tubulogenesis regulated by the transcription factor SOX9[J]. Gastroenterology, 2009, 136(7): 2325-2333. DOI:10.1053/j.gastro.2009.02.051.
[17]KOBAYASHI T, MIZUNO H, IMAYOSHI I, et al. The cyclic gene Hes1 contributes to diverse differentiation responses of embryonic stem cells[J]. Gene Develop, 2009, 23(16): 1870-1875. DOI:10.1101/gad.1823109.
[18]KAKINUMA S, NAKAUCHI H, WATANABE M. Hepatic stem/progenitor cells and stemcell transplantation for the treatment of liver disease[J]. J Gastroenterol, 2009, 44(3): 167-172. DOI:10.1007/s005350082297z.

相似文献/References:

[1]罗茜,王斌,雷增杰,等.肝星状细胞具有向肝细胞样细胞分化的干细胞潜能[J].第三军医大学学报,2013,35(07):604.
 Luo Xi,Wang Bin,Lei Zengjie,et al.Hepatic stellate cells might possess potential capabilities of stem cells to differentiate into hepatocyte-like cells in vitro[J].J Third Mil Med Univ,2013,35(09):604.
[2]呙琳琳,戴立里,唐静.丹参素对PDGF-BB刺激下肝星状细胞的抑制作用[J].第三军医大学学报,2011,33(15):1610.
 Guo Linlin,Dai Lili,Tang Jing.Salvianic acid A (Danshensu) inhibits proliferation and activation in rat hepatic stellate cells induced by platelet derived growth factor-BB[J].J Third Mil Med Univ,2011,33(09):1610.
[3]王军民,姚希贤,杨书良.丹参对AngⅡ刺激鼠HSCs活化Ca2+效应抑制作用的随机对照试验[J].第三军医大学学报,2005,27(08):756.
[4]刘戈云,何文华,李弼民,等.熊果酸对瘦素诱导的肝星状细胞JAK2-STAT3活化及活性氧产生的影响[J].第三军医大学学报,2011,33(19):2016.
 Liu Geyun,He Wenhua,Li Bimin,et al.Effect of ursolic acid on leptin-induced JAK2-STAT3 activation and reactive oxygen species generation in hepatic stellate cells[J].J Third Mil Med Univ,2011,33(09):2016.
[5]武鹏宇,戴立里,唐静,等.骨形态发生蛋白-7对人肝星状细胞转化生长因子β信号转导的影响[J].第三军医大学学报,2010,32(13):1433.
 Wu Pengyu,Dai Lili,Tang Jing,et al.Effect of bone morphogenetic protein-7 on transforming growth factor β signaling in hepatic stellate cells[J].J Third Mil Med Univ,2010,32(09):1433.
[6]叶平,杨波,唐文,等.p38MAPK信号通路对乙醛刺激的大鼠肝星状细胞增殖、凋亡及细胞周期的影响[J].第三军医大学学报,2010,32(20):2197.
 Ye Ping,Yang Bo,Tang Wen,et al.p38MAPK signal pathway modulates proliferation, apoptosis and cell cycle of rat hepatic stellate cells stimulated by acetaldehyde[J].J Third Mil Med Univ,2010,32(09):2197.
[7]唐静,戴立里,呙琳琳,等.肝纤维化中丹参素对TGFβ1/Smads/ERK信号通路的影响及其相互关系[J].第三军医大学学报,2011,33(11):1159.
 Tang Jing,Dai Lili,Guo Linlin,et al.Tanshinol inhibits TGFβ1/Smads/ERK signaling pathways in rat hepatic stellate cells[J].J Third Mil Med Univ,2011,33(09):1159.
[8]李冬,戴立里,余斌斌,等.丹参素对大鼠肝星状细胞增殖、凋亡及NF-κB活性的影响[J].第三军医大学学报,2009,31(08):724.
 LI Dong,DAI Li-li,YU Bin-bin,et al.Tanshinol induces proliferation, apoptosis and NF-κB activation in cultured rat hepatic stellate cells after IL-1β inducement[J].J Third Mil Med Univ,2009,31(09):724.
[9]覃浩,孙旭锐,欧仕玉,等.荔枝核总黄酮预防大鼠肝纤维化的初步研究[J].第三军医大学学报,2011,33(22):2353.
 Qin Hao,Sun Xurui,Ou Shiyu,et al.Total flavone from Litchi chinensis Sonn prevents hepatic fibrosis in rats[J].J Third Mil Med Univ,2011,33(09):2353.
[10]李俊峰,赵正斌,岳伟,等.维生素C和E对瘦素诱导的肝星状细胞增殖与TIMP-1上调的抑制作用[J].第三军医大学学报,2012,34(04):303.
 Li Junfeng,Zhao Zhengbin,Yue Wei,et al.Vitamins C and E suppress leptin-induced proliferation and TIMP-1 upregulation in hepatic stellate cells[J].J Third Mil Med Univ,2012,34(09):303.

更新日期/Last Update: 2018-05-10