CORTISTATIN REGULATES FIBROSIS AND MYOFIBROBLAST ACTIVATION IN
EXPERIMENTAL HEPATOTOXIC- AND CHOLESTATIC-INDUCED LIVER INJURY
Raquel Benitez1, Marta Caro1,
Eduardo Andres-Leon1, Francisco
O’Valle2, Mario Delgado1
1Institute of Parasitology and Biomedicine
Lopez-Neyra, PT Salud, Granada, Spain
2Dept. of Pathology, School of Medicine, IBIMER and
IBS-Granada, University of Granada, Spain
Authors’ emails: Raquel Benitez:rbenitez@ipb.csic.es; Marta
Caro: martacm@ipb.csic.es;
Eduardo Andres-Leon:eduardo.andres@csic.es;
Francisco O’Valle: fovalle@ugr.es;
Mario Delgado:mdelgado@ipb.csic.es
Corresponding author: Mario Delgado, Institute of Parasitology
and Biomedicine Lopez-Neyra IPBLN-CSIC, PT Salud, Granada, Spain. email:
mdelgado@ipb.csic.es. ORCID: 0000-0003-1893-5982.
Word count: 4893, Introduction (493 words), Results (1360
words), Discussion (1041 words).
Data availability. The data supporting the findings of this
study are available from the corresponding author. Raw fastq files for
RNAseq are publicly available at Sequence Read Archive
(SRA): PRJNA714069. Upregulated and downregulated DEGs (FDR
p-value<0.05) between CST-/- andCST+/+ HSCs are listed in Table S5.
Acknowledgments. We thank J. Campos-Salinas and A. Barroso
(IPBLN-CSIC) for their assistance in designing PCR and RNAseq strategy,
and L. de Lecea (Stanford University, CA, USA) for providing theCST-/- mice to establish initially the colony
of animals used in this study.
Competing interests. None declared.
Funding. This study was mainly supported by the Spanish
Ministry of Science and Innovation (MICINN, grant SAF2015-67787-R). R.B.
was recipient of FPI fellowship from MICINN and E.A-L. was recipient of
postdoctoral fellowship from regional Andalusian Government.
Author contributions. R.B. conducted most of the experiments
and analyzed all the data. M.C. carried out real-time PCRs. E.A-L.
analysed RNAseq results and public human data bases. F.O’V. performed
the histopathological studies. M.D. conceived and designed the study,
conducted in vivo experimental models, analyzed the data and
wrote the manuscript. All authors edited the manuscript.
Ethic statement. The experiments reported in this study
followed the ethical guidelines for investigations of experimental
animals approved by the Animal Care and Use board and the Ethical
Committee of Spanish Council of Scientific Research and performed in
accordance with the guidelines from Directive 2010/63/EU of the European
Parliament on the protection of animals used for scientific purposes.
Animal studies are reported in compliance with the ARRIVE guidelines and
with the recommendations made by the British Journal of
Pharmacology .
Declaration of transparency and scientific rigour. This
Declaration acknowledges that this paper adheres to the principles for
transparent reporting and scientific rigour of preclinical research as
stated in the BJP guidelines for Design & Analysis, Immunoblotting and
Immunochemistry and Animal Experimentation, and as recommended by
funding agencies, publishers and other organizations engaged with
supporting research.
Abbreviations: BDL: bile duct ligation; CST: cortistatin;CST+/+ : wild-type mice;CST+/- : partially cortistatin-deficient mice;CST-/- : totally cortistatin-deficient mice;
CTGF: connective tissue growth factor; DEG: differentially expressed
gene; ECM: extracellular matrix; FDR, false discovery rate; GFAP: glial
fibrillary acidic protein; GO: gene ontology; H&E: hematoxylin and
eosin; GHSR: growth hormone-secretagogue receptor; PF: periportal
fibroblast; RNAseq: RNA sequencing; RPLP0: ribosomal protein lateral
stalk subunit P0; αSMA: α-smooth muscle cell; sstr:
somatostatin-receptor.