Promotion of skeletal slow-twitch myofiber gene expression through miR-499-5p by procyanidin B2
The aim of this study was to investigate whether proanthocyanidins B2(PB2)regulate the expression of skeletal slow-twitch myofiber gene through miR-499-5p.After C2C12 cells and porcine skeletal muscle satellite cells were induced to differen-tiate into myotubes,the myotubes were treated with 0 μg/mL,2.5 μg/mL,5 μg/mL and 10 µg/mL PB2 to investigate the effects of PB2 on slow muscle fiber gene expression.In the mechanism studies,10 μg/mL of PB2 was added to C2C12 myotubes and porcine myotubes,respectively,while miR-499-5p expression was inhibited by transfection with 200 nM of miR-499-5p in-hibitor.The results showed that different doses of PB2 could promote the expression of MyHC Ⅰ gene in C2C12 myotubes(P＜0.05).The addition of 5 µg/mL and 10 μg/mL PB2 significantly increased the expression of MyHC Ⅰ gene in porcine myotubes(P＜0.05).The addition of 5 μg/mL and 10 μg/mL PB2 up-regulated the expression of miR-499-5p in C2C12 myotubes and porcine myotubes(P＜0.05).The mechanistic studies showed that inhibition of miR-499-5p attenuated the promotion of PB2 on miR-499-5p,MyHC I mRNA and slow MyHC protein.The above results demonstrated that PB2 could promote the expression of slow muscle fiber gene through miR-499-5p.
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