Ganoderma microsporum immunomodulatory protein, GMI, promotes C2C12 myoblast differentiation in vitro via upregulation of Tid1 and STAT3 acetylation.

Wan-Huai Teo,Yu-Ning Fan,Jeng-Fan Lo,Tung-Fu Huang,Chih-Yang Huang,Paul Lucas

doi:10.1371/journal.pone.0244791

Abstract

Ageing and chronic diseases lead to muscle loss and impair the regeneration of skeletal muscle. Thus, it’s crucial to seek for effective intervention to improve the muscle regeneration. Tid1, a mitochondrial co-chaperone, is important to maintain mitochondrial membrane potential and ATP synthesis. Previously, we demonstrated that mice with skeletal muscular specific Tid1 deficiency displayed muscular dystrophy and postnatal lethality. Tid1 can interact with STAT3 protein, which also plays an important role during myogenesis. In this study, we used GMI, immunomodulatory protein of Ganoderma microsporum, as an inducer in C2C12 myoblast differentiation. We observed that GMI pretreatment promoted the myogenic differentiation of C2C12 myoblasts. We also showed that the upregulation of mitochondria protein Tid1 with the GMI pre-treatment promoted myogenic differentiation ability of C2C12 cells. Strikingly, we observed the concomitant elevation of STAT3 acetylation (Ac-STAT3) during C2C12 myogenesis. Our study suggests that GMI promotes the myogenic differentiation through the activation of Tid1 and Ac-STAT3.

Highlights

Skeletal muscle, wrapped with the connective tissues, constitutes of myotubes that bundles into myofibrils
The cells were expanded in growth medium of Dulbecco’s Modified Eagle Medium (DMEM), 10% fetal bovine serum (FBS), 1% L-glutamine and 1% Penicillin-Streptomycin Amphotericin (PSA) at 37 ̊C under 5% CO2
Immunoblotting analyses demonstrated that the expression profile of those known myogenesis markers, including MyoD, myosin heavy chain (MyHC) and signal transducer and activator of transcription 3 (STAT3), which is similar to the previous findings [22] (Fig 1B)

Summary

Introduction

Skeletal muscle, wrapped with the connective tissues, constitutes of myotubes that bundles into myofibrils. It comprises about 40% to 45% of our body weight and enables our body to maintain the posture and to perform a wide range of movements, motions and stability [1,2]. Ageing or chronic diseases such as atrophy, cachexia and sarcopenia will lead to muscular loss and the deficiency to maintain or restore the normal structure and function of the impaired muscle [2,5,6]. Muscle regeneration and transplantation of myogenic cells is an essential therapy for the muscular dystrophies

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Results

Conclusion