Contribution of Mature Hepatocytes to Biliary Regeneration in Rats with Acute and Chronic Biliary Injury

Ya-Hui Chen; Hui-Ling Chen; Chin-Sung Chien; Shang-Hsin Wu; Yi-Tian Ho; Chun-Hsien Yu; Mei-Hwei Chang

doi:10.1371/journal.pone.0134327

Contribution of Mature Hepatocytes to Biliary Regeneration in Rats with Acute and Chronic Biliary Injury

PLoS One. 2015 Aug 26;10(8):e0134327. doi: 10.1371/journal.pone.0134327. eCollection 2015.

Authors

Ya-Hui Chen¹, Hui-Ling Chen², Chin-Sung Chien³, Shang-Hsin Wu³, Yi-Tian Ho², Chun-Hsien Yu⁴, Mei-Hwei Chang⁵

Affiliations

¹ Department of Pediatrics, Taipei Tzu-Chi Hospital, The Buddhist Tzuchi Medical Foundation, Taipei, Taiwan; Department of Pediatrics, Buddhist Tzu-Chi University College of Medicine, Hualien, Taiwan; Graduate Institute of Clinical Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan.
² Hepatitis Research Center, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan.
³ Graduate Institute of Clinical Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan.
⁴ Department of Pediatrics, Taipei Tzu-Chi Hospital, The Buddhist Tzuchi Medical Foundation, Taipei, Taiwan; Department of Pediatrics, Buddhist Tzu-Chi University College of Medicine, Hualien, Taiwan.
⁵ Graduate Institute of Clinical Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan; Hepatitis Research Center, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan; Department of Pediatrics, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan.

Abstract

Whether hepatocytes can convert into biliary epithelial cells (BECs) during biliary injury is much debated. To test this concept, we traced the fate of genetically labeled [dipeptidyl peptidase IV (DPPIV)-positive] hepatocytes in hepatocyte transplantation model following acute hepato-biliary injury induced by 4,4'-methylene-dianiline (DAPM) and D-galactosamine (DAPM+D-gal) and in DPPIV-chimeric liver model subjected to acute (DAPM+D-gal) or chronic biliary injury caused by DAPM and bile duct ligation (DAPM+BDL). In both models before biliary injury, BECs are uniformly DPPIV-deficient and proliferation of DPPIV-deficient hepatocytes is restricted by retrorsine. We found that mature hepatocytes underwent a stepwise conversion into BECs after biliary injury. In the hepatocyte transplantation model, DPPIV-positive hepatocytes entrapped periportally proliferated, and formed two-layered plates along portal veins. Within the two-layered plates, the hepatocytes gradually lost their hepatocytic identity, proceeded through an intermediate state, acquired a biliary phenotype, and subsequently formed bile ducts along the hilum-to-periphery axis. In DPPIV-chimeric liver model, periportal hepatocytes expressing hepatocyte nuclear factor-1β (HNF-1β) were exclusively DPPIV-positive and were in continuity to DPPIV-positives bile ducts. Inhibition of hepatocyte proliferation by additional doses of retrorsine in DPPIV-chimeric livers prevented the appearance of DPPIV-positive BECs after biliary injury. Moreover, enriched DPPIV-positive BEC/hepatic oval cell transplantation produced DPPIV-positive BECs or bile ducts in unexpectedly low frequency and in mid-lobular regions. These results together suggest that mature hepatocytes but not contaminating BECs/hepatic oval cells are the sources of periportal DPPIV-positive BECs. We conclude that mature hepatocytes contribute to biliary regeneration in the environment of acute and chronic biliary injury through a ductal plate configuration without the need of exogenously genetic or epigenetic manipulation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aniline Compounds / pharmacology
Animals
Bile Ducts / cytology
Bile Ducts / drug effects
Bile Ducts / injuries*
Bile Ducts / physiology*
Cell Transdifferentiation / drug effects
Cell Transplantation
Dipeptidyl Peptidase 4 / metabolism
Galactosamine / pharmacology
Hepatocytes / cytology*
Hepatocytes / drug effects
Hepatocytes / metabolism
Pyrrolizidine Alkaloids / pharmacology
Rats
Regeneration* / drug effects

Substances

Aniline Compounds
Pyrrolizidine Alkaloids
Galactosamine
Dipeptidyl Peptidase 4
4,4'-diaminodiphenylmethane
retrorsine

Grants and funding

This study was supported by grants from the National Science Council of the Republic of China (NSC97-2321-B-002-021-MY2 (HLC), NSC99-2314-B-002-041 (MHC), NSC99-2628-B-002-069-MY3 (HLC), NSC 101-2314-B-303 -022-1 (CHY), NSC102-2314-B-002-129 (HLC)), Taipei Tzu-Chi Hospital (TCRD-TPE-103-RT-8) (CHY), and the National Health Research Institute (NHRI-EX103-10001BI) (MHC). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.