Synthetic curcuminoid analogues abrogate oxidationinduced cell death and promote myogenic differentiation of C2C12 mouse myoblasts

Chittipong Tipbunjong,Yindee Kitiyanant,Apichart Suksamrarn,Vachiraporn Ajavakom,Chumpol Pholpramool,Piyawat Sookbangnop

doi:10.4314/tjpr.v17i8.4

Abstract

Purpose: To investigate the ability of two synthetic curcuminoid analogues, 6-(4-hydroxy-3-methoxyphenethyl)-5-(3-(4-hydroxy-3-methoxyphenyl)propanoyl)-4-phenyl-3,4-dihydropyrimidin-2(1H)-one (compound A) and 6-(4-hydroxy-3-methoxyphenethyl)-4-(4-hydroxy-3-methoxyphenyl)-5-(3-(4- hydroxy-3-methoxyphenyl)propanoyl)-3,4-dihydropyrimidin-2(1H)-one (compound B), to protect against oxidation-induced cell death and the potential to enhance proliferation and differentiation of C2C12 myoblast cells.Methods: Antioxidant activity of curcuminoid analogues was evaluated by DPPH assay. The cytotoxic activity of the compounds (0 - 25 mM) on C2C12 myoblasts was determined by MTT assay while the effect on cell proliferation was assessed by BrdU uptake. Myoblast cell differentiation was measured by the formation of myotubes and myosin heavy chain (MHC) protein expression using immunofluorescence staining and Western blotting, respectively.Results: Both curcuminoid analogues exhibited strong anti-oxidant activity of up to 3-fold greater than that of ascorbic acid, and were non-toxic to C2C12 myoblasts at concentrations up to 25 mM. Furthermore, these curcuminoid analogues mitigated myoblast cell death induced by oxidative stress. Notably, both analogues (10 nM) had no effect on cell proliferation. However, only compound A significantly enhanced myoblast differentiation comparable to the effects of dihydrotestosterone (1 μM) and estradiol (10 nM).Conclusion: The results suggest that compound A may serve as a lead compound for the development of suitable therapeutic agents for muscle injuries and diseases.Keywords: Curcuminoid analogues, Antioxidant, Cell proliferation, Cell differentiation, Myoblasts

Highlights

Skeletal muscle mass and strength are progressively loss with aging leading to the decline in functional ability known as sarcopenia [1]
Both curcuminoid analogues at concentrations up to 25 μM showed no effect on C2C12 myoblast cell viability determined by MTT assay
Based on MTT assay which detects the living, but not dead cells, these data indicate that the number of living myoblast cells in curcuminoid treatment groups was increased

Summary

Introduction

Skeletal muscle mass and strength are progressively loss with aging leading to the decline in functional ability known as sarcopenia [1]. -©---2-0--1--8---T--h-e---a--u-t-h--o-r-s--.--T-h--i-s--w--o--r-k--i-s--l-i-c-e--n-s--e--d--u--n--d-e--r--t-h-e---C--r-e--a-t-i-v-e---C--o--m--m--T-o-rn-o-sp--A-J-t-tP-r-ih-b-au--rt-imo--n-R--4e-.-s0-,--IAn--ut-eg--ru-n-sa-t-t-i2o--0n-1-a-8l--;L--1ic-7-e-(n-8-s-)-e:-1-4--8- 3 production of reactive oxygen species (ROS) is enhanced while the anti-oxidative capacity is diminished [2] resulting in cellular oxidative stress critical for cell death. Under this condition, the proliferation and differentiation capacities of satellite cells, which are required for muscle repair and/or regeneration, decrease [1] leading to a gradual decline in muscle mass and function. Dihydropyrimidin-2(1H)-one (compound A) and 6(4-hydroxy-3-methoxyphenethyl)-4-(4-hydroxy-3methoxyphenyl)-5-(3-(4-hydroxy-3methoxyphenyl)propanoyl)-3,4-dihydropyrimidin2(1H)-one (compound B), were synthesized and characterized as previously described [9]. Other biological properties of these compounds have not yet been investigated

Results

Discussion

Conclusion