To evaluate the effects of dietary supplementation with succinic acid on growth performance, flesh quality, glucose, and lipid metabolism of Nile tilapia (Oreochromis niloticus) fed a high-carbohydrate diet (HCD), five iso-nitrogenous and iso-lipidic diets were prepared as follows: HCD (control group) consisting of 55% corn starch and HCD supplemented with 0.5%, 1.0%, 2.0%, and 4.0% succinic acid, respectively. Tilapia with an initial body weight of 204.90 ± 1.23 g randomly assigned to 15 tanks with 3 replicates per group and 10 fish per tank fed for 8 weeks. Increasing dietary succinic acid supplementation resulted in significant second-order polynomial relationship in the weight gain rate (WGR), specific growth rate (SGR), feed conversion ratio (FCR), protein efficiency rate (PER), viscerosomatic index, condition factor, and contents of muscular crude lipid and glycogen (P < 0.05). The hepatosomatic index, mesenteric fat index, liver glycogen content and crude lipid contents of the whole-body and liver demonstrated significantly linear and second-order polynomial relationship (P < 0.05). Quadratic curve model analysis based on WGR, SGR, PER, and FCR demonstrated that optimal supplementation with succinic acid in the HCD of Nile tilapia ranged from 1.83% to 2.43%. Fish fed with 1.0% succinic acid had higher muscular hardness, increased the contents of alkali-soluble hydroxyproline in collagen, docosahexaenoic acid (DHA) and n-3 polyunsaturated fatty acid (n-3PUFA) in muscle, and lower total fatty acid content in muscle (P < 0.05) compared with the control group. Compared to the control group, dietary supplementation with 1.0% succinic acid significantly increased the contents of total bounding amino acid (arginine, histidine, isoleucine, lysine, methionine, alanine, proline), total flavor amino acid (free aspartic acid), the catalase (CAT) activity and total antioxidant capacity, and the mRNA relative expression levels of CAT, superoxide dismutase (SOD), and nuclearfactor erythroidderived 2-like 2 (Nrf2) in muscle (P < 0.05). Furthermore, succinic acid supplementation significantly up-regulated mRNA relative expression levels of glycolysis genes (hexokinase 2 [HK2], phosphofructokinase, muscle-A [PFKMA], and phosphofructokinase, muscle-B [PFKMB]), a key glycogen synthesis gene (glycogen synthase [GYS]), and lipid catabolism genes (carnitine palmitoyltransferase-1B [CPT1B], hormone sensitive lipase [HSL], and lipoprotein lipase [LPL]), while down-regulating the mRNA relative expression level of fatty acid synthase (FASN) in muscle (P < 0.05). In conclusion, dietary supplementation with 1.83% to 2.43% succinic acid improved muscle quality by increasing muscle antioxidant capacity and hardness, changing muscle amino acid and fatty acid composition, and regulating muscle glucose and lipid metabolism.