Knockout of secretin ameliorates biliary and liver phenotypes during alcohol-induced hepatotoxicity

Konstantina Kyritsi; Nan Wu; Tianhao Zhou; Guido Carpino; Leonardo Baiocchi; Lindsey Kennedy; Lixian Chen; Ludovica Ceci; Alison Ann Meyer; Nipuni Barupala; Antonio Franchitto; Paolo Onori; Burcin Ekser; Eugenio Gaudio; Chaodong Wu; Corinn Marakovits; Sanjukta Chakraborty; Heather Francis; Shannon Glaser; Gianfranco Alpini

doi:10.1186/s13578-022-00945-w

Knockout of secretin ameliorates biliary and liver phenotypes during alcohol-induced hepatotoxicity

Cell Biosci. 2023 Jan 9;13(1):5. doi: 10.1186/s13578-022-00945-w.

Authors

Konstantina Kyritsi^#¹, Nan Wu^#¹, Tianhao Zhou^#¹, Guido Carpino², Leonardo Baiocchi³, Lindsey Kennedy^{1

4}, Lixian Chen¹, Ludovica Ceci^{1

2}, Alison Ann Meyer¹, Nipuni Barupala¹, Antonio Franchitto⁵, Paolo Onori², Burcin Ekser⁶, Eugenio Gaudio², Chaodong Wu⁷, Corinn Marakovits¹, Sanjukta Chakraborty⁸, Heather Francis^{9

10}, Shannon Glaser¹¹, Gianfranco Alpini^{12

13}

Affiliations

¹ Division of Gastroenterology and Hepatology, Department of Medicine, Indiana University School of Medicine, Indianapolis, IN, USA.
² Department of Anatomical, Histological, Forensic Medicine and Orthopedics Sciences, La Sapienza University of Rome, Rome, Italy.
³ Unit of Hepatology, Tor Vergata University, Rome, Italy.
⁴ Division of Research, Indiana Center for Liver Research, Gastroenterology, Medicine, Richard L. Roudebush VA Medical Center and Indiana University, 702 Rotary Circle, Rm. 013C, Indianapolis, IN, 46202-2859, USA.
⁵ Department of Movement, Human and Health Sciences, University of Rome "Foro Italico", Rome, Italy.
⁶ Division of Transplant Surgery, Department of Surgery, Indiana University, Indianapolis, IN, USA.
⁷ Department of Nutrition, Texas A&M University, College Station, TX, USA.
⁸ Department of Medical Physiology, Texas A&M University School of Medicine, 8447 Riverside Parkway, MREB II, Room 2342, Bryan, TX, 77807-3260, USA.
⁹ Division of Gastroenterology and Hepatology, Department of Medicine, Indiana University School of Medicine, Indianapolis, IN, USA. heafranc@iu.edu.
¹⁰ Division of Research, Indiana Center for Liver Research, Gastroenterology, Medicine, Richard L. Roudebush VA Medical Center and Indiana University, 702 Rotary Circle, Rm. 013C, Indianapolis, IN, 46202-2859, USA. heafranc@iu.edu.
¹¹ Department of Medical Physiology, Texas A&M University School of Medicine, 8447 Riverside Parkway, MREB II, Room 2342, Bryan, TX, 77807-3260, USA. sglaser@tamu.edu.
¹² Division of Gastroenterology and Hepatology, Department of Medicine, Indiana University School of Medicine, Indianapolis, IN, USA. galpini@iu.edu.
¹³ Division of Research, Indiana Center for Liver Research, Gastroenterology, Medicine, Richard L. Roudebush VA Medical Center and Indiana University, 702 Rotary Circle, Rm. 013C, Indianapolis, IN, 46202-2859, USA. galpini@iu.edu.

^# Contributed equally.

Abstract

Background: Alcohol-related liver disease (ALD) is characterized by ductular reaction (DR), liver inflammation, steatosis, fibrosis, and cirrhosis. The secretin (Sct)/secretin receptor (SR) axis (expressed only by cholangiocytes) regulates liver phenotypes in cholestasis. We evaluated the role of Sct signaling on ALD phenotypes.

Methods: We used male wild-type and Sct^-/- mice fed a control diet (CD) or ethanol (EtOH) for 8 wk. Changes in liver phenotypes were measured in mice, female/male healthy controls, and patients with alcoholic cirrhosis. Since Cyp4a10 and Cyp4a11/22 regulate EtOH liver metabolism, we measured their expression in mouse/human liver. We evaluated: (i) the immunoreactivity of the lipogenesis enzyme elongation of very-long-chain fatty acids 1 (Elovl, mainly expressed by hepatocytes) in mouse/human liver sections by immunostaining; (ii) the expression of miR-125b (that is downregulated in cholestasis by Sct) in mouse liver by qPCR; and (iii) total bile acid (BA) levels in mouse liver by enzymatic assay, and the mRNA expression of genes regulating BA synthesis (cholesterol 7a-hydroxylase, Cyp27a1, 12a-hydroxylase, Cyp8b1, and oxysterol 7a-hydroxylase, Cyp7b11) and transport (bile salt export pump, Bsep, Na⁺-taurocholate cotransporting polypeptide, NTCP, and the organic solute transporter alpha (OSTa) in mouse liver by qPCR.

Results: In EtOH-fed WT mice there was increased biliary and liver damage compared to control mice, but decreased miR-125b expression, phenotypes that were blunted in EtOH-fed Sct^-/- mice. The expression of Cyp4a10 increased in cholangiocytes and hepatocytes from EtOH-fed WT compared to control mice but decreased in EtOH-fed Sct^-/- mice. There was increased immunoreactivity of Cyp4a11/22 in patients with alcoholic cirrhosis compared to controls. The expression of miR-125b decreased in EtOH-fed WT mice but returned at normal values in EtOH-fed Sct^-/- mice. Elovl1 immunoreactivity increased in patients with alcoholic cirrhosis compared to controls. There was no difference in BA levels between WT mice fed CD or EtOH; BA levels decreased in EtOH-fed Sct^-/- compared to EtOH-fed WT mice. There was increased expression of Cyp27a1, Cyp8b1, Cyp7b1, Bsep, NTCP and Osta in total liver from EtOH-fed WT compared to control mice, which decreased in EtOH-fed Sct^-/- compared to EtOH-fed WT mice.

Conclusions: Targeting Sct/SR signaling may be important for modulating ALD phenotypes.

Keywords: Biliary senescence; Ductular reaction; Fatty liver diseases; Hepatic steatosis; Lipogenesis.

Abstract

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