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Knockout of α-calcitonin gene-related peptide attenuates cholestatic liver injury by differentially regulating cellular senescence of hepatic stellate cells and cholangiocytes

Laboratory Investigation (2019) | Download Citation

Abstract

α-Calcitonin gene-related peptide (α-CGRP) is a 37-amino acid neuropeptide involved in several pathophysiological processes. α-CGRP is involved in the regulation of cholangiocyte proliferation during cholestasis. In this study, we aimed to evaluate if α-CGRP regulates bile duct ligation (BDL)-induced liver fibrosis by using a α-CGRP knockout (α-CGRP-/-) mouse model. α-CGRP-/- and wild-type (WT) mice were subjected to sham surgery or BDL for 7 days. Then, liver fibrosis and cellular senescence as well as the expression of kinase such as p38 and C-Jun N-terminal protein kinase (JNK) in mitogen-activated protein kinases (MAPK) signaling pathway were evaluated in total liver, together with measurement of cellular senescence in cholangiocytes or hepatic stellate cells (HSCs). There was enhanced hepatic expression of Calca (coding α-CGRP) and the CGRP receptor components (CRLR, RAMP-1 and RCP) in BDL and in both WT α-CGRP-/- and BDL α-CGRP-/- mice, respectively. Moreover, there was increased CGRP serum levels and hepatic mRNA expression of CALCA and CGRP receptor components in late-stage PSC samples compared to healthy control samples. Depletion of α-CGRP reduced liver injury and fibrosis in BDL mice that was associated with enhanced cellular senescence of hepatic stellate cells and reduced senescence of cholangiocytes as well as decreased activation of p38 and JNK MAPK signaling pathway. Cholangiocyte supernatant from BDL α-CGRP-/- mice inhibited the activation and increased cellular senescence of cultured human HSCs (HHSCs) compared to HHSCs stimulated with BDL cholangiocyte supernatant. Taken together, endogenous α-CGRP promoted BDL-induced cholestatic liver fibrosis through differential changes in senescence of HSCs and cholangiocytes and activation of p38 and JNK signaling. Modulation of α-CGRP/CGRP receptor signaling may be key for the management of biliary senescence and liver fibrosis in cholangiopathies.

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Present address for Drs. Bernuzzi and Invernizzi: Program for Autoimmune Liver Diseases, International Center for Digestive Health, Department of Medicine and Surgery, University of Milan-Bicocca, Milano, Italy.

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Acknowledgements

This work was supported by the Dr. Nicholas C. Hightower Centennial Chair of Gastroenterology from Baylor Scott & White, a VA Research Career Scientist Award and a VA Merit award to Dr. Alpini (5I01BX000574), a VA Merit Award (1I01BX001724) to Dr. Meng, and the NIH grants DK058411, DK076898, DK107310, DK110035, DK062975, AA025997, and AA025157 to Drs. Alpini, Meng, and Glaser. Portions of this work were supported by (i) a VA Merit Award (1I01BX003031) from the United States Department of Veteran’s affairs, Biomedical Laboratory Research and Development Service and an R01 from NIH NIDDK (DK108959) to Dr. Francis (ii) and a Nature Science foundation of China (No.81873563) to Dr. Wan. This material is the result of work supported with resources and the use of facilities at the Central Texas Veterans Health Care System, Temple, Texas.

Author information

Author notes

  1. Ying Wan and Ludovica Ceci share the first authorship.

  2. Drs. Gianfranco Alpini, Fanyin Meng, and Shannon Glaser share the senior authorship.

Affiliations

  1. Department of Pathophysiology, Southwest Medical University, Luzhou, 646000, China

    • Ying Wan
  2. Department of Medical Physiology, Texas A&M University Health Science Center, Temple, TX, USA

    • Ludovica Ceci
    • , Nan Wu
    • , Tianhao Zhou
    • , Lixian Chen
    • , Julie Venter
    • , Heather Francis
    • , Konstantina Kyritsi
    • , Paul Baker
    • , Gianfranco Alpini
    • , Fanyin Meng
    •  & Shannon Glaser
  3. Research, Central Texas Veterans Health Care System, Temple, TX, USA

    • Heather Francis
    • , Gianfranco Alpini
    •  & Fanyin Meng
  4. Digestive Disease Research Center, Baylor Scott & White Health, Temple, TX, USA

    • Heather Francis
    • , Gianfranco Alpini
    • , Fanyin Meng
    •  & Shannon Glaser
  5. Humanitas Clinical and Research Center, Rozzano (MI), Italy

    • Francesca Bernuzzi
    •  & Pietro Invernizzi
  6. Department of Pathophysiology, Key Lab for Shock and Microcirculation Research of Guangdong Province, Southern Medical University, Guangzhou, China

    • Qiaobing Huang
  7. Department of Nutrition and Food Science, Texas A&M University, College Station, TX, 77843, USA

    • Chaodong Wu
  8. Department of Pathology, Microbiology, and Immunology, Vanderbilt University School of Medicine, Nashville, TN, USA

    • Amelia Sybenga
  9. Research Foundation, Baylor Scott & White Health, Temple, TX, USA

    • Fanyin Meng

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Conflict of interest

The authors declare that they have no conflict of interest.

Corresponding author

Correspondence to Gianfranco Alpini.

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DOI

https://doi.org/10.1038/s41374-018-0178-5