Damage to the myogenic differentiation of C2C12 cells by heat stress is associated with up-regulation of several selenoproteins

Jiayong Tang,Gang Jia,Xiaoling Chen,Guangmang Liu,Aihua He,Hui Yan,Jingyi Cai,Haiying Shang,Gang Tian,Hua Zhao

doi:10.1038/s41598-018-29012-6

Jiayong Tang, Gang Jia + Show 8 more

Open Access

https://doi.org/10.1038/s41598-018-29012-6

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Abstract

This study was conducted to profile the selenoprotein encoding genes or proteins in mouse C2C12 cells and integrate their roles in the skeletal cell damage induced by heat stress (HS). Cells were cultured at 37.0 °C or 41.5 °C for 4, 6 or 8 days. The mRNA expression of 24 selenoprotein encoding genes and abundance of 5 selenoproteins were investigated. HS suppressed myogenic differentiation and impaired the development of muscle myotubes. HS down-regulated (P < 0.01) mRNA abundance of MYOD and MYOGENIN, and decreased (P < 0.01) MYOGENIN protein expression, HS elevated (P < 0.01) HSP70 and (P < 0.01) the ratio of BCL-2 to BAX at both mRNA and protein level. Meanwhile, HS up-regulated (P < 0.01–0.05) expressions of 18, 11 and 8 selenoprotein encoding genes after 4, 6 and 8 days of hyperthermia, and only down-regulated (P < 0.01) DIO2 after 6 and 8 days of hyperthermia, respectively. Furthermore, HS influenced expression of selenoproteins and up-regulated (P < 0.01–0.05) GPX1, GPX4 and SEPN1 after 6 days of HS. The damage to development of mouse skeletal muscle myotubes by HS accompanied with the up-regulation of both selenoprotein encoding genes and proteins, which suggested a potential protective effect of selenoprotein on hyperthermia associated damage in C2C12 cells.

Highlights

The climate change with increased surface temperature on the earth occurs globally in the past decades
We further investigated effect of heat stress (HS) on mRNA and protein expression of MYOD, MYOGENIN, AMPKα1 and AMPKα2 in the differentiating C2C12 cells (Fig. 2)
Compared to the CK groups, HS decreased (P < 0.01) mRNA expression of MYOD by 66%, 60%, 83%, and MYOGENIN by 66%, 63%, 83%, in C2C12 cells at day 4, 6, 8, respectively (Fig. 2A–C)

Summary

Introduction

The climate change with increased surface temperature on the earth occurs globally in the past decades. During differentiation of C2C12 cells, myoblasts undergo remodeling to form mature myotubes in parallel with the increased expression of muscle specific genes[16]. C2C12 isolated from mouse lines by Yaffe and Saxel, which mimics the development of skeletal muscle in vivo, representing an excellent model to study myogenic regulation and response to stimuli[14,31]. We conducted this to determine (1) impact of HS on myogenic differentiation in C2C12 cells; (2) effect of HS on the gene or protein expression of selenoproteins, myoblast differentiation-related protein, apoptosis-related protein and HSP70; (3) impact of HS on antioxidant attributes of C2C12 cells.

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