title: Secretin stimulates biliary cell proliferation by regulating expression of microRNA 125b and microRNA let7a in mice.
abstract: OBJECTIVE
  Proliferating cholangiocytes secrete and respond to neuroendocrine hormones, including secretin. We investigated whether secretin secreted by S cells and cholangiocytes stimulates biliary proliferation in mice.
  
  
  METHODS
  Cholestasis was induced in secretin knockout (Sct(-/-)) and wild-type (control) mice by bile duct ligation (BDL). At days 3 and 7 after BDL, control and Sct(-/-) mice received tail-vein injections of morpholinos against microRNA 125b or let7a. One week later, liver tissues and cholangiocytes were collected. Immunohistochemical, immunoblot, luciferase reporter, and real-time polymerase chain reaction assays were performed. Intrahepatic bile duct mass (IBDM) and proliferation were measured. Secretin secretion was measured in conditioned media from cholangiocytes and S cells and in serum and bile.
  
  
  RESULTS
  Secretin secretion was increased in supernatants from cholangiocytes and S cells and in serum and bile after BDL in control mice. BDL Sct(-/-) mice had lower IBDM, reduced proliferation, and reduced production of vascular endothelial growth factor (VEGF) A and nerve growth factor (NGF) compared with BDL control. BDL and control mice given morpholinos against microRNA 125b or let7a had increased IBDM. Livers of mice given morpholinos against microRNA 125b had increased expression of VEGFA, and those treated with morpholinos against microRNA let7a had increased expression of NGF. Secretin regulated VEGF and NGF expression that negatively correlated with microRNA 125b and let7a levels in liver tissue.
  
  
  CONCLUSIONS
  After liver injury, secretin produced by cholangiocytes and S cells reduces microRNA 125b and let7a levels, resulting in up-regulation of VEGF and NGF. Modulation of cholangiocyte expression of secretin could be a therapeutic approach for biliary diseases.
authors.affiliation: Research, Central Texas Veterans Health Care System, Temple, Texas; Scott & White Digestive Disease Research Center, Scott & White, Temple, Texas; Department of Medicine, Division of Gastroenterology, Texas A&M Health Science Center College of Medicine, Temple, Texas.
authors.forename: Shannon
authors.initials: S
authors.lastname: Glaser
authors.affiliation: Research, Central Texas Veterans Health Care System, Temple, Texas; Scott & White Digestive Disease Research Center, Scott & White, Temple, Texas; Department of Medicine, Division of Gastroenterology, Texas A&M Health Science Center College of Medicine, Temple, Texas; Academic Operations, Scott & White, Temple, Texas.
authors.forename: Fanyin
authors.initials: F
authors.lastname: Meng
authors.affiliation: Department of Medicine, Division of Gastroenterology, Texas A&M Health Science Center College of Medicine, Temple, Texas.
authors.forename: Yuyan
authors.initials: Y
authors.lastname: Han
authors.affiliation: Department of Anatomical, Histological, Forensic Medicine, and Orthopedics Sciences, Sapienza, Rome, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Onori
authors.affiliation: School of Biological Sciences, The University of Hong Kong, Hong Kong, China.
authors.forename: Billy K
authors.initials: BK
authors.lastname: Chow
authors.affiliation: Research, Central Texas Veterans Health Care System, Temple, Texas; Scott & White Digestive Disease Research Center, Scott & White, Temple, Texas; Department of Medicine, Division of Gastroenterology, Texas A&M Health Science Center College of Medicine, Temple, Texas; Academic Operations, Scott & White, Temple, Texas.
authors.forename: Heather
authors.initials: H
authors.lastname: Francis
authors.affiliation: Department of Medicine, Division of Gastroenterology, Texas A&M Health Science Center College of Medicine, Temple, Texas.
authors.forename: Julie
authors.initials: J
authors.lastname: Venter
authors.affiliation: Research, Central Texas Veterans Health Care System, Temple, Texas.
authors.forename: Kelly
authors.initials: K
authors.lastname: McDaniel
authors.affiliation: Department of Medicine, Universita' Politecnica delle Marche, Ancona, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Marzioni
authors.affiliation: Liver Unit and Center for Autoimmune Liver Diseases, Humanitas Clinical and Research Center, Rozzano, Milan, Italy.
authors.forename: Pietro
authors.initials: P
authors.lastname: Invernizzi
authors.affiliation: Division of Gastroenterology, Yamagata University Faculty of Medicine, Yamagata, Japan.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Ueno
authors.affiliation: Department of Hepatobiliary Surgery, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
authors.forename: Jia-ming
authors.initials: JM
authors.lastname: Lai
authors.affiliation: Department of Hepatobiliary Surgery, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
authors.forename: Li
authors.initials: L
authors.lastname: Huang
authors.affiliation: Research, Central Texas Veterans Health Care System, Temple, Texas.
authors.forename: Holly
authors.initials: H
authors.lastname: Standeford
authors.affiliation: Department of Medico-Surgical Sciences and Biotechnologies, Polo Pontino, Italy.
authors.forename: Domenico
authors.initials: D
authors.lastname: Alvaro
authors.affiliation: Department of Anatomical, Histological, Forensic Medicine, and Orthopedics Sciences, Sapienza, Rome, Italy.
authors.forename: Eugenio
authors.initials: E
authors.lastname: Gaudio
authors.affiliation: Department of Anatomical, Histological, Forensic Medicine, and Orthopedics Sciences, Sapienza, Rome, Italy; Eleonora Lorillard Spencer Cenci Foundation, Rome, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Franchitto
authors.affiliation: Research, Central Texas Veterans Health Care System, Temple, Texas; Scott & White Digestive Disease Research Center, Scott & White, Temple, Texas; Department of Medicine, Division of Gastroenterology, Texas A&M Health Science Center College of Medicine, Temple, Texas. Electronic address: galpini@tamu.edu.
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Alpini
mesh_terms: D000818:Animals; D017209:Apoptosis; D001646:Bile / Q000378:metabolism; D001652:Bile Ducts / Q000378:metabolism* / Q000473:pathology; D049109:Cell Proliferation; D002478:Cells, Cultured; D002779:Cholestasis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D017077:Culture Media, Conditioned / Q000378:metabolism; D004195:Disease Models, Animal; D019858:Enteroendocrine Cells / Q000378:metabolism; D008099:Liver / Q000378:metabolism* / Q000473:pathology; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D060172:Morpholinos / Q000008:administration & dosage; D020932:Nerve Growth Factor / Q000378:metabolism; D012633:Secretin / Q000097:blood / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; D035683:MicroRNAs; C527468:Mirn125 microRNA, mouse; D060172:Morpholinos; D042461:Vascular Endothelial Growth Factor A; C542687:mirnlet7 microRNA, mouse; C467480:vascular endothelial growth factor A, mouse; D012633:Secretin; D020932:Nerve Growth Factor
keywords: Biliary Epithelium; Gastrointestinal Hormones; Heterogeneity; cAMP