Establishment of a mouse model of vascular endothelial-mesenchymal transdifferentiation genetic tracing and its role in liver fibrosis studies

Hao XU; Bai RUAN; Zhiwen LI; Zhiqiang FANG; Lin WANG; Kefeng DOU

doi:10.3969/j.issn.1001-5256.2022.04.018

Volume 38 Issue 4

Apr. 2022

Turn off MathJax

Article Contents

Article Navigation > Journal of Clinical Hepatology > 2022 > 38(4): 832-836

PDF( 4480 KB)

Establishment of a mouse model of vascular endothelial-mesenchymal transdifferentiation genetic tracing and its role in liver fibrosis studies

DOI: 10.3969/j.issn.1001-5256.2022.04.018

Hao XU¹,
Bai RUAN^{1, 2},
Zhiwen LI¹,
Zhiqiang FANG¹,
Lin WANG¹,
Kefeng DOU^{1
,
,
,}

1.
Department of Hepatobiliary, Pancreatic and Splenic Surgery, Xijing Hospital, Air Force Medical University, Xi'an 710032, China
2.
Center of Clinical Aerospace Medicine, School of Aerospace Medicine, Air Force Medical University, Xi'an 710032, China

Research funding:

The National Key Research and Development Program of China (2016YFA0102100);

National Natural Science Foundation of China (81770560);

National Natural Science Foundation of China (81800533)

More Information

Corresponding author: DOU Kefeng, doukef@fmmu.edu.cn (ORCID: 0000-0002-8857-5663)
Received Date: 2021-10-26
Accepted Date: 2021-11-28
Published Date: 2022-04-20

Abstract

Abstract

Objective To establish Cdh5-Cre^ERT/Acta2-tdTomato-STOP^floxed-eGFP knockin genetic tracing mice, and to investigate its application in studies on vascular endothelial cell transition in liver fibrosis. Methods Cdh5-Cre^ERT mice were mated with Acta2-KI mice, and the Cdh5-Cre^ERT/Acta2-KI genetic tracing mice were obtained and identified by PCR genotyping. Primary liver sinusoid endothelial cells (LSECs) were isolated and cultured, and a model of CCl₄-induced liver fibrosis was established. LSECs and liver tissue were collected for immunofluorescent staining to observe the expression of the fluorescent proteins tdTomato and eGFP. Results After being induced by tamoxifen, LSECs and liver tissue of Cdh5-Cre^ERT/Acta2-KI genetic tracing mice expressed eGFP under the conditions for epithelial-mesenchymal transdifferentiation established in vivo and in vitro, while the control group without induction expressed tdTomato alone. Conclusion The successfully established Cdh5-Cre^ERT/Acta2-KI genetic tracing mice can realize the effective labeling of epithelial-mesenchymal transdifferentiation, which provides a genetic tracing basis for the diverse sources of mesenchymal myofibroblasts in liver fibrosis.
- Liver Cirrhosis,
- Epithelial-Mesenchymal Transition,
- Endothelial Cells,
- Mice

FullText(HTML)

References(15)

References

[1]	LEE UE, FRIEDMAN SL. Mechanisms of hepatic fibrogenesis[J]. Best Pract Res Clin Gastroenterol, 2011, 25(2): 195-206. DOI: 10.1016/j.bpg.2011.02.005.
[2]	MEDERACKE I, HSU CC, TROEGER JS, et al. Fate tracing reveals hepatic stellate cells as dominant contributors to liver fibrosis independent of its aetiology[J]. Nat Commun, 2013, 4: 2823. DOI: 10.1038/ncomms3823.
[3]	RAY K. Liver: hepatic stellate cells hold the key to liver fibrosis[J]. Nat Rev Gastroenterol Hepatol, 2014, 11(2): 74. DOI: 10.1038/nrgastro.2013.244.
[4]	PIERA-VELAZQUEZ S, MENDOZA FA, JIMENEZ SA. Endothelial to mesenchymal transition (EndoMT) in the pathogenesis of human fibrotic diseases[J]. J Clin Med, 2016, 5(4): 45. DOI: 10.3390/jcm5040045.
[5]	SUN X, NKENNOR B, MASTIKHINA O, et al. Endothelium-mediated contributions to fibrosis[J]. Semin Cell Dev Biol, 2020, 101: 78-86. DOI: 10.1016/j.semcdb.2019.10.015.
[6]	RUAN B, DUAN JL, XU H, et al. Capillarized liver sinusoidal endothelial cells undergo partial endothelial-mesenchymal transition to actively deposit sinusoidal ECM in liver fibrosis[J]. Front Cell Dev Biol, 2021, 9: 671081. DOI: 10.3389/fcell.2021.671081.
[7]	THOMSON JG, RUCKER EB 3rd, PIEDRAHITA JA. Mutational analysis of loxP sites for efficient Cre-mediated insertion into genomic DNA[J]. Genesis, 2003, 36(3): 162-167. DOI: 10.1002/gene.10211.
[8]	KOS CH. Cre/loxP system for generating tissue-specific knockout mouse models[J]. Nutr Rev, 2004, 62(6 Pt 1): 243-246. DOI: 10.1301/nr2004.jun243-246.
[9]	JIANG ZJ, YUE ZS, YANG YC, et al. Improvement method of isolation of mouse liver Sinusoidal endothelial cell[J]. Prog Mod Biomed, 2018, 18(6): 1034-1039. DOI: 10.13241/j.cnki.pmb.2018.06.007. 蒋子剑, 岳振生, 杨毅聪, 等. 小鼠肝血窦内皮细胞分离与鉴定新方法[J]. 现代生物医学进展, 2018, 18(6): 1034-1039. DOI: 10.13241/j.cnki.pmb.2018.06.007.
[10]	WANG S, FRIEDMAN SL. Hepatic fibrosis: A convergent response to liver injury that is reversible[J]. J Hepatol, 2020, 73(1): 210-211. DOI: 10.1016/j.jhep.2020.03.011.
[11]	TERKELSEN MK, BENDIXEN SM, HANSEN D, et al. Transcriptional dynamics of hepatic sinusoid-associated cells after liver injury[J]. Hepatology, 2020, 72(6): 2119-2133. DOI: 10.1002/hep.31215.
[12]	TRAUTWEIN C, FRIEDMAN SL, SCHUPPAN D, et al. Hepatic fibrosis: Concept to treatment[J]. J Hepatol, 2015, 62(1 Suppl): S15-S24. DOI: 10.1016/j.jhep.2015.02.039.
[13]	ZEISBERG EM, TARNAVSKI O, ZEISBERG M, et al. Endothelial-to-mesenchymal transition contributes to cardiac fibrosis[J]. Nat Med, 2007, 13(8): 952-961. DOI: 10.1038/nm1613.
[14]	ZEISBERG EM, POTENTA SE, SUGIMOTO H, et al. Fibroblasts in kidney fibrosis emerge via endothelial-to-mesenchymal transition[J]. J Am Soc Nephrol, 2008, 19(12): 2282-2287. DOI: 10.1681/ASN.2008050513.
[15]	HASHIMOTO N, PHAN SH, IMAIZUMI K, et al. Endothelial-mesenchymal transition in bleomycin-induced pulmonary fibrosis[J]. Am J Respir Cell Mol Biol, 2010, 43(2): 161-172. DOI: 10.1165/rcmb.2009-0031OC.

Relative Articles

Supplements(0)

Cited By

Proportional views

Proportional views

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Figures(4) / Tables(1)

Get Citation

PDF

XML

Article Metrics

Article views (899) PDF downloads(57)

Establishment of a mouse model of vascular endothelial-mesenchymal transdifferentiation genetic tracing and its role in liver fibrosis studies

DOI: 10.3969/j.issn.1001-5256.2022.04.018

Abstract

References

Proportional views

Catalog

通讯作者: 陈斌, bchen63@163.com

Article Metrics

Proportional views

Related

Establishment of a mouse model of vascular endothelial-mesenchymal transdifferentiation genetic tracing and its role in liver fibrosis studies

DOI: 10.3969/j.issn.1001-5256.2022.04.018

Abstract

References

Proportional views

Catalog

通讯作者: 陈斌, bchen63@163.com

Article Metrics

Proportional views

Related

About Journal

Submission Guideline

Journal Online

Hepatobiliary College

Guidelines

Export File

Citation

Format

Content