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Molecules 2016, 21(8), 997; doi:10.3390/molecules21080997

Potential Application of p-Coumaric Acid on Differentiation of C2C12 Skeletal Muscle and 3T3-L1 Preadipocytes—An in Vitro and in Silico Approach

1
Grassland and Forage Division, National Institute of Animal Science, Rural Development Administration, Cheonan 330-801, Korea
2
Laboratory of Animal Physiology, Graduate School of Agricultural Science, Tohoku University, Aoba, Sendai 980-8577, Japan
3
Department of Botany and Microbiology, Addiriyah Chair for Environmental Studies, College of Science, King Saud University, Riyadh 11451, Saudi Arabia
4
Department of Oriental Medicine Materials, Dongsin University, Naju 520-714, Korea
5
Research Center of Bioactive Materials, Institute of Molecular Biology and Genetics, Chonbuk National University, Jeonju 561-756, Korea
6
Biocenter, Gyeonggi Institute of Science and Technology, Suwon 443-270, Korea
7
Department of Bioinformatics, Marudupandiyar College, Tamilnadu 613-403, India
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editor: Derek J. McPhee
Received: 22 June 2016 / Revised: 26 July 2016 / Accepted: 27 July 2016 / Published: 2 August 2016
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Abstract

Coumaric acid (CA) is a phenolic acid of the hydroxycinnamic acid family, and it has many biological functions such as anti-oxidant, anti-inflammatory, antidiabetic, anti-ulcer, anti-platelet, anti-cancer activities, etc. In the present study, we planned to analyse the potential molecular function of CA on skeletal muscle and preadipocytes differentiation using PCR and Western blot techniques. First, we analysed the impact of CA on C2C12 skeletal muscle differentiation. It revealed that CA treatment inhibited horse serum-induced skeletal muscle differentiation as evidenced by the decreased expression of early myogenic differentiation markers such as Myogenin and myoD via the AMP activated protein kinase- alpha AMPK-α mediated pathway. Furthermore, the level of lipid accumulation and changes in genes and protein expressions that are associated with lipogenesis and lipolysis were analyzed in 3T3-L1 cells. The Oil Red O staining evidenced that CA treatment inhibited lipid accumulation at the concentration of 0.1 and 0.2 mM. Furthermore, coumaric acid treatment decreased the expression of main transcriptional factors such as CCAAT/enhancer binding protein-alpha (C/EBP-α) and peroxisome proliferator-activated receptor gamma-2 (PPAR-γ2). Subsequently, CA treatment decreased the expression of sterol regulatory element binding protein-1 (SREBP-1), fatty acid synthase (FAS), acetyl CoA carboxylase (ACC) and adiponectin. Finally, we identified conformational changes induced by CA in PPAR-γ2 using computational biology tools. It revealed that CA might downregulate the PPAR-γ2 expression by directly binding with amino acids of PPAR-γ2 by hydrogen at 3.26 distance and hydrophobic interactions at 3.90 contact distances. These data indicated that CA suppressed skeletal muscle and preadipocytes differentiation through downregulation of the main transcriptional factors and their downstream targets. View Full-Text
Keywords: coumaric acid; C2C12 and 3T3-L1cells; AMPK-α; PPAR-γ2; PCR and Western blot; computational biology coumaric acid; C2C12 and 3T3-L1cells; AMPK-α; PPAR-γ2; PCR and Western blot; computational biology
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MDPI and ACS Style

Ilavenil, S.; Kim, D.H.; Srigopalram, S.; Arasu, M.V.; Lee, K.D.; Lee, J.C.; Lee, J.S.; Renganathan, S.; Choi, K.C. Potential Application of p-Coumaric Acid on Differentiation of C2C12 Skeletal Muscle and 3T3-L1 Preadipocytes—An in Vitro and in Silico Approach. Molecules 2016, 21, 997.

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