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

艾麦提·牙森; 金鑫; 陈梓昕; 王伟; 李德卫

doi:10.16016/j.1000-5404.201711122

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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

基础医学 | 更新时间：2026-01-09

- 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
- TGF-β1 signaling mediated hepatic stellate cells promote hepatic progenitor cells differentiation into cholangiocytes
- 第三军医大学学报 2018年40卷第9期页码：746-752
- 作者机构：
  
  400016 重庆, 重庆医科大学附属第一医院肝胆外科, 实验研究中心
- 作者简介：
  
  [李德卫, E-mail:lidewei406@sina.com LI Dewei, E-mail:lidewei406@sina.com
- 基金信息：
  
  国家自然科学基金面上项目(81470898)
- DOI：10.16016/j.1000-5404.201711122
  中图分类号： R322.47;R329-33;R329.26
- 收稿：2017-11-15，
  
  修回：2018-02-28，
  
  纸质出版：2018-05-15
- 稿件说明：
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艾麦提·牙森, 金鑫, 陈梓昕, 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(9):746-752.

AI MAITI·Ya Sen, Xin JIN, Zixin CHEN, et al. TGF-β1 signaling mediated hepatic stellate cells promote hepatic progenitor cells differentiation into cholangiocytes[J]. Journal of Third Military Medical University, 2018, 40(9): 746-752.
艾麦提·牙森, 金鑫, 陈梓昕, 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(9):746-752. DOI： 10.16016/j.1000-5404.201711122.

AI MAITI·Ya Sen, Xin JIN, Zixin CHEN, et al. TGF-β1 signaling mediated hepatic stellate cells promote hepatic progenitor cells differentiation into cholangiocytes[J]. Journal of Third Military Medical University, 2018, 40(9): 746-752. DOI： 10.16016/j.1000-5404.201711122.

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(

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(

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 (

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 (

0.01).

Conclusion

TGF-β1-activated hepatic stellate cells induce hepatic progenitor cells differentiation into cholangiocytes.

关键词

Keywords

references

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