Inhibitor of FTO, Rhein, Restrains the Differentiation of Myoblasts and Delays Skeletal Muscle Regeneration

N6-methyladenosine (m6A) is a crucial RNA modification affecting skeletal muscle development. Rhein, an anti-inflammatory extract, inhibits <i>FTO</i>, a key demethylase in m6A metabolism. Our study showed that during muscle fiber formation, <i>FTO</i> and <i>ALKBH5</i> expression increased while m6A levels decreased. After muscle injury, <i>FTO</i> and <i>ALKBH5</i> expression initially rose but later fell, while m6A levels initially dropped and then recovered. Inhibition of <i>FTO</i> by Rhein reduced MyHC and <i>MyoG</i> expression, indicating myoblast differentiation suppression. In a mouse model, Rhein decreased MyHC expression and muscle fiber cross-sectional area, delaying muscle regeneration. Rhein's ability to increase RNA m6A modification delays skeletal muscle remodeling post-injury, suggesting a new medicinal application for this plant extract.