title: Glial cell line-derived neurotrophic factor-induced mice liver defatting: A novel strategy to enable transplantation of steatotic livers.
abstract: Moderate macrovesicular steatosis (>30%), which is present in almost 50% of livers considered for transplantation, increases the risk of primary graft dysfunction. Our previously published data showed that glial cell line-derived neurotrophic factor (GDNF) is protective against high-fat diet (HFD)-induced hepatic steatosis in mice. Hence, we hypothesized that perfusion of steatotic livers with GDNF may reduce liver fat content before transplantation. Livers from 8 weeks of regular diet (RD) and of HFD-fed mice were perfused ex vivo for 4 hours with either vehicle, GDNF, or a previously described defatting cocktail. The liver's residual fat was quantified colorimetrically using a triglyceride (TG) assay kit and by Oil Red O (ORO) and Nile red/Hoechst staining. Liver tissue injury was assessed by using a lactate dehydrogenase (LDH) activity assay. In vitro induction of lipolysis in HepG2 cells was assessed by measuring glycerol and free fatty acid release. ORO staining showed significantly more steatosis in livers from HFD-fed mice compared with RD-fed mice (P < 0.001). HFD livers perfused with GDNF had significantly less steatosis than those not perfused (P = 0.001) or perfused with vehicle (P < 0.05). GDNF is equally effective in steatotic liver defatting compared to the defatting cocktail; however, GDNF induces less liver damage than the defatting cocktail. These observations were consistent with data obtained from assessment of liver TG content. Assessment of liver injury revealed significant hepatocyte injury in livers perfused with the control defatting cocktail but no evidence of injury in livers perfused with either GDNF or vehicle. In vitro, GDNF reduced TG accumulation in HepG2 cells and stimulated increased TG lipolysis. In conclusion, GDNF can decrease mice liver fat content to an acceptable range and could be a potential defatting agent before liver transplantation.
authors.affiliation: Division of Digestive Diseases, Emory University School of Medicine, Atlanta, GA.|Atlanta VA Medical Center, Decatur, GA.
authors.forename: Sahar
authors.initials: S
authors.lastname: Taba Taba Vakili
authors.affiliation: Division of Digestive Diseases, Emory University School of Medicine, Atlanta, GA.
authors.forename: Roshni
authors.initials: R
authors.lastname: Kailar
authors.affiliation: Division of Digestive Diseases, Emory University School of Medicine, Atlanta, GA.
authors.forename: Khalidur
authors.initials: K
authors.lastname: Rahman
authors.affiliation: Division of Digestive Diseases, Emory University School of Medicine, Atlanta, GA.|Atlanta VA Medical Center, Decatur, GA.
authors.forename: Behtash Ghazi
authors.initials: BG
authors.lastname: Nezami
authors.affiliation: Division of Digestive Diseases, Emory University School of Medicine, Atlanta, GA.|Atlanta VA Medical Center, Decatur, GA.
authors.forename: Simon Musyoka
authors.initials: SM
authors.lastname: Mwangi
authors.affiliation: Division of Digestive Diseases, Emory University School of Medicine, Atlanta, GA.|Atlanta VA Medical Center, Decatur, GA.
authors.forename: Frank A
authors.initials: FA
authors.lastname: Anania
authors.affiliation: Division of Digestive Diseases, Emory University School of Medicine, Atlanta, GA.|Atlanta VA Medical Center, Decatur, GA.
authors.forename: Shanthi
authors.initials: S
authors.lastname: Srinivasan
mesh_terms: D000818:Animals; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology / Q000378:metabolism; D003124:Colorimetry; D059305:Diet, High-Fat / Q000009:adverse effects; D004195:Disease Models, Animal; D005234:Fatty Liver / Q000209:etiology / Q000628:therapy*; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000009:adverse effects / Q000627:therapeutic use*; D006085:Graft Survival / Q000187:drug effects; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D008066:Lipolysis / Q000187:drug effects; D016031:Liver Transplantation / Q000379:methods*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010477:Perfusion; D055031:Primary Graft Dysfunction / Q000517:prevention & control*; D051381:Rats; D014280:Triglycerides / Q000032:analysis / Q000378:metabolism*
chemical_list: C495041:Gdnf protein, rat; D051100:Glial Cell Line-Derived Neurotrophic Factor; D014280:Triglycerides