PUFA Treatment Affects C2C12 Myocyte Differentiation, Myogenesis Related Genes and Energy Metabolism.

Marua Abu Risha,Siriluck Ponsuksili,Puntita Siengdee,Dirk Dannenberger,Klaus Wimmers

doi:10.3390/genes12020192

Marua Abu Risha, Siriluck Ponsuksili + Show 3 more

Open Access

https://doi.org/10.3390/genes12020192

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Abstract

Polyunsaturated fatty acids (PUFAs) are the main components of cell membrane affecting its fluidity, signaling processes and play a vital role in muscle cell development. The effects of docosahexaenoic acid (DHA) on myogenesis are well known, while the effects of arachidonic acid (AA) are largely unclear. The purpose of this study is to evaluate the effect of two PUFAs (DHA and AA) on cell fate during myogenic processes, Wnt signaling and energy metabolism by using the C2C12 cells. The cells were treated with different concentrations of AA or DHA for 48 h during the differentiation period. PUFA treatment increased mRNA level of myogenic factor 5 (Myf5), which is involved in early stage of myoblast proliferation. Additionally, PUFA treatment prevented myoblast differentiation, indicated by decreased myotube fusion index and differentiation index in parallel with reduced mRNA levels of myogenin (MyoG). After PUFA withdrawal, some changes in cell morphology and myosin heavy chain mRNA levels were still observed. Expression of genes associated with Wnt signaling pathway, and energy metabolism changed in PUFA treatment in a dose and time dependent manner. Our data suggests that PUFAs affect the transition of C2C12 cells from proliferation to differentiation phase by prolonging proliferation and preventing differentiation.

Highlights

Myogenesis is a complex process involving many events including proliferation, differentiation and migration of myoblasts, and is regulated by several signaling pathways such as Wnt, BMP and Shh [1,2]
Cell membranes consist of two layers which are primarily composed of phospholipids and contain cholesterols and sphingolipids [8]
The xCelligence Real Time Cell Analyzer (RTCA) system records cell impedance as a function of cell density and was used to monitor cell viability of C2C12 cells treated with polyunsaturated fatty acids (PUFAs) (DHA or arachidonic acid (AA))

Summary

Introduction

Myogenesis is a complex process involving many events including proliferation, differentiation and migration of myoblasts, and is regulated by several signaling pathways such as Wnt, BMP and Shh [1,2]. Myogenic regulatory factors (MRFs) are a part of this complex network and regulate embryonic myogenesis, drive muscle regeneration, and repair damage by activating satellite cells. Myf regulates the final differentiation of myotubes [6] Besides this complex network, cell membrane properties (receptors, channels, lipid composition and polarity) are considered strong players in myogenesis [7] and key elements for maintaining cell physiology and cell survival. Polyunsaturated fatty acids (PUFAs), long fatty acids with more than one double bond, can change the functionality of membranes Both n-3-PUFAs (ω-3) and n-6-PUFAs (ω-6) are considered important molecular components regulating various functions such as anti-inflammatory and pro-inflammatory properties [9,10]. EPA and DHA pose an inhibitory effect on myoblast proliferation and differentiation, and downregulate muscle-related gene expression [14]

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