Cyclic Stretch Facilitates Myogenesis in C2C12 Myoblasts and Rescues Thiazolidinedione-Inhibited Myotube Formation

Ya-Ju Chang; Yun-Ju Chen; Chia-Wei Huang; Shih-Chen Fan; Bu-Miin Huang; Wen-Tsan Chang; Yau-Sheng Tsai; Fong-Chin Su; Chia-Ching Wu

doi:10.3389/fbioe.2016.00027

Cyclic Stretch Facilitates Myogenesis in C2C12 Myoblasts and Rescues Thiazolidinedione-Inhibited Myotube Formation

Front Bioeng Biotechnol. 2016 Mar 21:4:27. doi: 10.3389/fbioe.2016.00027. eCollection 2016.

Authors

Ya-Ju Chang¹, Yun-Ju Chen², Chia-Wei Huang³, Shih-Chen Fan⁴, Bu-Miin Huang⁵, Wen-Tsan Chang⁶, Yau-Sheng Tsai⁷, Fong-Chin Su⁸, Chia-Ching Wu⁹

Affiliations

¹ Department of Cell Biology and Anatomy, National Cheng Kung University, Tainan, Taiwan; Institute of Basic Medical Sciences, National Cheng Kung University, Tainan, Taiwan.
² Department of Microbiology, College of Medicine, National Taiwan University , Taipei , Taiwan.
³ Department of Cell Biology and Anatomy, National Cheng Kung University, Tainan, Taiwan; Institute of Basic Medical Sciences, National Cheng Kung University, Tainan, Taiwan; Department of Occupational Therapy, National Cheng Kung University, Tainan, Taiwan.
⁴ Department of Occupational Therapy, I-Shou University , Kaohsiung , Taiwan.
⁵ Department of Cell Biology and Anatomy, National Cheng Kung University , Tainan , Taiwan.
⁶ Department of Biochemistry and Molecular Biology, National Cheng Kung University , Tainan , Taiwan.
⁷ Institute of Clinical Medicine, National Cheng Kung University , Tainan , Taiwan.
⁸ Department of Biomedical Engineering, National Cheng Kung University, Tainan, Taiwan; Medical Device Innovation Center, National Cheng Kung University, Tainan, Taiwan.
⁹ Department of Cell Biology and Anatomy, National Cheng Kung University, Tainan, Taiwan; Institute of Basic Medical Sciences, National Cheng Kung University, Tainan, Taiwan; Institute of Clinical Medicine, National Cheng Kung University, Tainan, Taiwan; Department of Biomedical Engineering, National Cheng Kung University, Tainan, Taiwan; Medical Device Innovation Center, National Cheng Kung University, Tainan, Taiwan.

Abstract

Thiazolidinedione (TZD), a specific peroxisome proliferator-activated receptor γ (PPARγ) agonist, was developed to control blood glucose in diabetes patients. However, several side effects were reported that increased the risk of heart failure. We used C2C12 myoblasts to investigate the role of PPARs and their transcriptional activity during myotube formation. The role of mechanical stretch during myogenesis was also explored by applying cyclic stretch to the differentiating C2C12 myoblasts with 10% strain deformation at 1 Hz. The myogenesis medium (MM), composed of Dulbecco's modified Eagle's medium with 2% horse serum, facilitated myotube formation with increased myosin heavy chain and α-smooth muscle actin (α-SMA) protein expression. The PPARγ protein and PPAR response element (PPRE) promoter activity decreased during MM induction. Cyclic stretch further facilitated the myogenesis in MM with increased α-SMA and decreased PPARγ protein expression and inhibited PPRE promoter activity. Adding a PPARγ agonist (TZD) to the MM stopped the myogenesis and restored the PPRE promoter activity, whereas a PPARγ antagonist (GW9662) significantly increased the myotube number and length. During the myogenesis induction, application of cyclic stretch rescued the inhibitory effects of TZD. These results provide novel perspectives for mechanical stretch to interplay and rescue the dysfunction of myogenesis with the involvement of PPARγ and its target drugs.

Keywords: C2C12; PPAR; myogenesis; myotube formation; stretch.