Sains Malaysiana 52(1)(2023): 47-56

http://doi.org/10.17576/jsm-2023-5201-04

Hyperglycaemia Attenuated C2C12 Myoblast Proliferation and Induced Skeletal Muscle Atrophy via Modulating Myogenic Regulatory Factors Genes Expression in Diabetic Rats

(Hiperglisemia Dilemahkan C2C12 Pembiakan Mioblas dan Atrofi Otot Rangka Terinduksi melalui Modulasi Faktor Kawalaturan Miogenesis Ekspresi Gen dalam Tikus Diabetis)

CHITTIPONG TIPBUNJONG^1,2,*, WIPAPAN KHIMMAKTONG¹, CHUMPOL PHOLPRAMOOL³ & PIYAPORN SURINLERT⁴

¹Division of Health and Applied Sciences, Faculty of Science, Prince of Songkla University, Songkhla 90110, Thailand

²Gut Biology and Microbiota Research Unit, Prince of Songkla University, Songkhla 90110, Thailand

³Department of Physiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand 

⁴Chulabhorn International College of Medicine, Thammasat University, Pathum Thani 12120, Thailand 

Received: 18 January 2022/Accepted: 3 November 2022

Abstract

Diabetes mellitus is characterised by high blood glucose level termed hyperglycaemia (HG). It has been reported to affect skeletal muscle by inducing skeletal muscle atrophy and skeletal muscle protein degradation leading to impairment of muscle function. This study aimed to investigate the effects of HG on the expression of myogenic regulatory factor genes in muscle progenitor cells and in skeletal muscle. The number of C2C12 myoblasts cultured in HG condition was significantly decreased compared to control in dose and time dependent manner. In addition, the number of Ki-67 positive nuclei was significantly decreased after treatment under HG condition. Real time PCR showed significant suppression of MyoD and myogenin, while Myf5 gene expression was significantly enhanced, compared to control. Furthermore, histological examinations of muscle fibres showed atrophy of tibialis anterior (TA) muscle from diabetic rats. The frequency of distribution of muscle fibre cross-sectional area (MCA) in diabetic rats was shifted leftward from that of normal control rats. In contrast, the MyoD and myogenin expression in TA muscle of diabetic rats were significantly increased compared to normal control rats. This study provides novel knowledge on the changing myogenic regulatory factor gene expression in hyperglycaemic condition, both in vitro and in vivo, leading to skeletal muscle atrophy.

Keywords: Atrophy; diabetes; myoblast; myogenesis; skeletal muscle

Abstrak

Diabetes mellitus dicirikan oleh tahap glukosa darah yang tinggi dipanggil hiperglisemia (HG). Ia telah dilaporkan menjejaskan otot rangka dengan mengaruh atrofi otot rangka dan degradasi protein otot rangka yang membawa kepada kemerosotan fungsi otot. Kajian ini bertujuan untuk mengkaji kesan HG ke atas pengekspresan gen faktor pengawalaturan miogenik dalam sel progenitor otot dan dalam otot rangka. Bilangan mioblas C2C12 yang dibiakkan dalam keadaan HG berkurangan dengan ketara berbanding dengan kawalan yang bergantung kepada dos dan masa. Di samping itu, bilangan nukleus positif Ki-67 berkurangan dengan ketara selepas rawatan dalam keadaan HG. PCR masa nyata menunjukkan penindasan signifikan MyoD dan miogenin, manakala pengekspresan gen Myf5 meningkat dengan signifikan, berbanding kawalan. Tambahan pula, pemeriksaan histologi gentian otot mendedahkan atrofi otot tibialis anterior (TA) bagi tikus diabetes. Kekerapan taburan kawasan keratan rentas gentian otot (MCA) pada tikus diabetes dianjak ke kiri berbanding tikus kawalan normal. Sebaliknya, pengekspresan MyoD dan miogenin dalam otot TA tikus diabetes meningkat dengan ketara berbanding tikus kawalan normal. Kajian ini memberikan pengetahuan baharu tentang perubahan pengekspresan gen faktor pengawalaturan miogenik dalam keadaan hiperglisemik yang membawa kepada atrofi otot rangka bagi kedua-dua keadaan in vitro dan in vivo.

Kata kunci: Atrofi; diabetes; mioblas; miogenesis; otot rangka

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^*Corresponding author; email: chittipong.t@psu.ac.th