PMID- 36174267 OWN - NLM STAT- MEDLINE DCOM- 20221108 LR - 20221108 IS - 1525-3171 (Electronic) IS - 0032-5791 (Print) IS - 0032-5791 (Linking) VI - 101 IP - 11 DP - 2022 Nov TI - Osteocalcin reduces fat accumulation and inflammatory reaction by inhibiting ROS-JNK signal pathway in chicken embryonic hepatocytes. PG - 102026 LID - S0032-5791(22)00317-0 [pii] LID - 10.1016/j.psj.2022.102026 [doi] LID - 102026 AB - Osteocalcin (OCN) has a function in preventing fatty liver hemorrhagic syndrome (FLHS) in poultry. The aim of this study was to investigate the effects of OCN on fat emulsion stimulated chicken embryonic hepatocytes and related signaling pathways. The primary chicken embryonic hepatocytes were isolated from the incubated 15-day (E15) pathogen free eggs and cultured with dulbecco's modified eagle medium (DMEM). After the hepatocyte density reached 80%, the cells were divided into 5 groups: control group (CONT), fat emulsion group (FE, 10% FE, v/v), FE with ucOCN at 1 ng/mL (FE-LOCN), 3 ng/mL (FE-MOCN), and 9 ng/mL (FE-HOCN). In addition, 2 mM N-Acetyl-L-cysteine (NAC) a reactive oxygen species (ROS) scavenger, and 5 muM SP600125, a Jun N-terminal kinase (JNK) inhibitor, were added separately in to the DMEM with 10% FE to test effects of FE on the function of ROS-JNK signal pathway. The number of hepatocytes, cell ultra-microstructure, viability, and apoptosis were detected after 48 h treatment, and the protein expressions and enzyme concentrations were detected after 72 h treatment. The results showed that, compared to the control group, FE increased the triglyceride (TG) concentration and lipid droplets (LDs) in chicken embryonic hepatocytes (P < 0.05), and induced hepatocytic edema with obviously mitochondrial swelling, membrane damage, and cristae rupture. FE also decreased ATP concentration, increased ROS concentrations and mitochondrial DNA (mtDNA) copy number, promoted inflammatory interleukin-1 (IL-1), IL-6, tumor necrosis factor-alpha (TNF-alpha) concentrations and hepatocytic apoptosis rate, and raised phospho-c-Jun N-terminal kinase (p-JNK) protein expressions. Compared to the FE group, ucOCN significantly increased hepatocyte viability, reduced hepatocytic TG concentrations and LDs numbers, and alleviated hepatocytic edema and mitochondrial swelling. Furthermore, ucOCN significantly decreased ROS concentrations, increased ATP concentrations, reduced IL-1, IL-6, TNF-alpha concentrations and hepatocytic apoptosis rate, and inhibited p-JNK protein expressions (P < 0.05). NAC had the similar functions of ucOCN reduced the ROS concentration and inhibited the TNF-alpha protein expression and p-JNK/JNK ration. Similarly, SP600125 reduced p-JNK/JNK protein expression, IL-1, IL-6, TNF-alpha, and TG concentrations without effects on ROS concentration and hepatocytic apoptosis. These results suggest that ucOCN alleviates FE-induced mitochondrial damage, cellular edema, and apoptosis of hepatocytes. These results reveal that the functions of ucOCN in reducing fat accumulation and inflammatory reaction in chicken embryonic hepatocytes are mostly via inhibiting the ROS-JNK signal pathway. CI - Copyright (c) 2022 The Authors. Published by Elsevier Inc. All rights reserved. FAU - Zhang, M AU - Zhang M AD - Joint International Research Laboratory of Animal Health and Animal Food Safety, College of Veterinary Medicine, Southwest University, Chongqing, 400715, China. FAU - Tu, W J AU - Tu WJ AD - Joint International Research Laboratory of Animal Health and Animal Food Safety, College of Veterinary Medicine, Southwest University, Chongqing, 400715, China. FAU - Zhang, Q AU - Zhang Q AD - Joint International Research Laboratory of Animal Health and Animal Food Safety, College of Veterinary Medicine, Southwest University, Chongqing, 400715, China. FAU - Wu, X L AU - Wu XL AD - Joint International Research Laboratory of Animal Health and Animal Food Safety, College of Veterinary Medicine, Southwest University, Chongqing, 400715, China. FAU - Zou, X Y AU - Zou XY AD - Joint International Research Laboratory of Animal Health and Animal Food Safety, College of Veterinary Medicine, Southwest University, Chongqing, 400715, China. FAU - Jiang, S AU - Jiang S AD - Joint International Research Laboratory of Animal Health and Animal Food Safety, College of Veterinary Medicine, Southwest University, Chongqing, 400715, China; Immunology Research Center, Medical Research Institute, Southwest University, Chongqing, 402460, China. Electronic address: jiangsha0527@swu.edu.cn. LA - eng PT - Journal Article DEP - 20220624 PL - England TA - Poult Sci JT - Poultry science JID - 0401150 RN - 1TW30Y2766 (pyrazolanthrone) RN - 0 (Reactive Oxygen Species) RN - 0 (Tumor Necrosis Factor-alpha) RN - 104982-03-8 (Osteocalcin) RN - 0 (Interleukin-6) RN - 0 (Emulsions) RN - EC 2.7.11.24 (JNK Mitogen-Activated Protein Kinases) RN - 0 (Interleukin-1) RN - 8L70Q75FXE (Adenosine Triphosphate) SB - IM MH - Chick Embryo MH - Animals MH - *Hepatocytes MH - Reactive Oxygen Species/metabolism MH - *Tumor Necrosis Factor-alpha MH - Chickens/metabolism MH - Osteocalcin/pharmacology MH - Interleukin-6/metabolism MH - Emulsions MH - Signal Transduction MH - JNK Mitogen-Activated Protein Kinases/metabolism MH - Apoptosis MH - Inflammation/veterinary/metabolism MH - Interleukin-1/metabolism/pharmacology MH - Adenosine Triphosphate/metabolism PMC - PMC9519800 OTO - NOTNLM OT - ROS-JNK OT - chicken embryonic hepatocyte OT - fat accumulation OT - fatty liver hemorrhagic syndrome OT - inflammatory reaction OT - osteocalcin EDAT- 2022/09/30 06:00 MHDA- 2022/11/09 06:00 PMCR- 2022/06/24 CRDT- 2022/09/29 18:14 PHST- 2022/04/21 00:00 [received] PHST- 2022/06/16 00:00 [revised] PHST- 2022/06/18 00:00 [accepted] PHST- 2022/09/30 06:00 [pubmed] PHST- 2022/11/09 06:00 [medline] PHST- 2022/09/29 18:14 [entrez] PHST- 2022/06/24 00:00 [pmc-release] AID - S0032-5791(22)00317-0 [pii] AID - 102026 [pii] AID - 10.1016/j.psj.2022.102026 [doi] PST - ppublish SO - Poult Sci. 2022 Nov;101(11):102026. doi: 10.1016/j.psj.2022.102026. Epub 2022 Jun 24.