PPAR γ/Nnat/NF-κB Axis Involved in Promoting Effects of Adiponectin on Preadipocyte Differentiation

A previous study has demonstrated that adiponectin (APN) could promote preadipocyte differentiation, and the present study further explored its mechanism. 3T3-L1 cells were infected with adenovirus holding human adiponectin gene apM1 and mouse neuronatin (Nnat) shRNA and initiated differentiation wh...

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Main Authors: Wenkai Yang, Wenjin Yuan, Xinghua Peng, Meiling Wang, Jie Xiao, Cheng Wu, Lie Luo
Format: Article
Language:English
Published: Wiley 2019-01-01
Series:Mediators of Inflammation
Online Access:http://dx.doi.org/10.1155/2019/5618023
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author Wenkai Yang
Wenjin Yuan
Xinghua Peng
Meiling Wang
Jie Xiao
Cheng Wu
Lie Luo
author_facet Wenkai Yang
Wenjin Yuan
Xinghua Peng
Meiling Wang
Jie Xiao
Cheng Wu
Lie Luo
author_sort Wenkai Yang
collection DOAJ
description A previous study has demonstrated that adiponectin (APN) could promote preadipocyte differentiation, and the present study further explored its mechanism. 3T3-L1 cells were infected with adenovirus holding human adiponectin gene apM1 and mouse neuronatin (Nnat) shRNA and initiated differentiation while coculturing with mature adipocytes stimulated with LPS. After 8 days, preadipocyte differentiation was observed by Oil Red O staining. Real-time quantitative PCR was used to evaluate mRNA expression levels of monocyte chemoattractant protein-1 (MCP-1), interleukin- (IL-) 6, IL-8, and tumor necrosis factor α (TNF-α). The levels of reactive oxygen species (ROS), total antioxidant capacity (T-AOC), malondialdehyde (MDA), and superoxide dismutase (SOD) in 3T3-L1 cells were detected. Western blotting was done to quantify the protein expression levels of Nnat, peroxisome proliferator-activated receptor (PPAR) γ, p65, and inhibitor of nuclear factor κB (IκB) α. Results demonstrated that APN overexpression markedly increased preadipocyte differentiation; inhibited gene expression of MCP-1, IL-6, IL-8, and TNF-α; reduced ROS and MDA release; increased T-AOC and SOD levels; upregulated Nnat, PPAR γ, and IκB α protein expressions; and downregulated p65 protein expression under LPS stimulation. However, the effects of APN were markedly attenuated when Nnat expression was knocked down. Taken together, the present study provided evidences that the effects of APN on promoting preadipocyte differentiation under inflammatory conditions via anti-inflammation and antioxidative stress may be regulated by the PPAR γ/Nnat/NF-κB signaling pathway.
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spelling doaj-art-97c2d111f51344b7bd5ca16edf24e8192025-08-20T03:37:08ZengWileyMediators of Inflammation0962-93511466-18612019-01-01201910.1155/2019/56180235618023PPAR γ/Nnat/NF-κB Axis Involved in Promoting Effects of Adiponectin on Preadipocyte DifferentiationWenkai Yang0Wenjin Yuan1Xinghua Peng2Meiling Wang3Jie Xiao4Cheng Wu5Lie Luo6Department of Cardiovascular Surgery, Affiliated Central People’s Hospital of Zhanjiang of Guangdong Medical University, Zhanjiang 524045, ChinaDepartment of Cardiovascular Medicine, Ganzhou People’s Hospital, Ganzhou 341000, ChinaDepartment of Cardio-Thoracic Surgery, Ganzhou People’s Hospital, Ganzhou 341000, ChinaDepartment of Cardio-Thoracic Surgery, Ganzhou People’s Hospital, Ganzhou 341000, ChinaDepartment of Cardio-Thoracic Surgery, Ganzhou People’s Hospital, Ganzhou 341000, ChinaDepartment of Cardio-Thoracic Surgery, Ganzhou People’s Hospital, Ganzhou 341000, ChinaDepartment of Cardio-Thoracic Surgery, Ganzhou People’s Hospital, Ganzhou 341000, ChinaA previous study has demonstrated that adiponectin (APN) could promote preadipocyte differentiation, and the present study further explored its mechanism. 3T3-L1 cells were infected with adenovirus holding human adiponectin gene apM1 and mouse neuronatin (Nnat) shRNA and initiated differentiation while coculturing with mature adipocytes stimulated with LPS. After 8 days, preadipocyte differentiation was observed by Oil Red O staining. Real-time quantitative PCR was used to evaluate mRNA expression levels of monocyte chemoattractant protein-1 (MCP-1), interleukin- (IL-) 6, IL-8, and tumor necrosis factor α (TNF-α). The levels of reactive oxygen species (ROS), total antioxidant capacity (T-AOC), malondialdehyde (MDA), and superoxide dismutase (SOD) in 3T3-L1 cells were detected. Western blotting was done to quantify the protein expression levels of Nnat, peroxisome proliferator-activated receptor (PPAR) γ, p65, and inhibitor of nuclear factor κB (IκB) α. Results demonstrated that APN overexpression markedly increased preadipocyte differentiation; inhibited gene expression of MCP-1, IL-6, IL-8, and TNF-α; reduced ROS and MDA release; increased T-AOC and SOD levels; upregulated Nnat, PPAR γ, and IκB α protein expressions; and downregulated p65 protein expression under LPS stimulation. However, the effects of APN were markedly attenuated when Nnat expression was knocked down. Taken together, the present study provided evidences that the effects of APN on promoting preadipocyte differentiation under inflammatory conditions via anti-inflammation and antioxidative stress may be regulated by the PPAR γ/Nnat/NF-κB signaling pathway.http://dx.doi.org/10.1155/2019/5618023
spellingShingle Wenkai Yang
Wenjin Yuan
Xinghua Peng
Meiling Wang
Jie Xiao
Cheng Wu
Lie Luo
PPAR γ/Nnat/NF-κB Axis Involved in Promoting Effects of Adiponectin on Preadipocyte Differentiation
Mediators of Inflammation
title PPAR γ/Nnat/NF-κB Axis Involved in Promoting Effects of Adiponectin on Preadipocyte Differentiation
title_full PPAR γ/Nnat/NF-κB Axis Involved in Promoting Effects of Adiponectin on Preadipocyte Differentiation
title_fullStr PPAR γ/Nnat/NF-κB Axis Involved in Promoting Effects of Adiponectin on Preadipocyte Differentiation
title_full_unstemmed PPAR γ/Nnat/NF-κB Axis Involved in Promoting Effects of Adiponectin on Preadipocyte Differentiation
title_short PPAR γ/Nnat/NF-κB Axis Involved in Promoting Effects of Adiponectin on Preadipocyte Differentiation
title_sort ppar γ nnat nf κb axis involved in promoting effects of adiponectin on preadipocyte differentiation
url http://dx.doi.org/10.1155/2019/5618023
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