The objective of the present research was to assess the influence of inositol supplementation on growth performance, histological morphology of liver, immunity and expression of immune-related genes in juvenile hybrid grouper (♀ Epinephelus fuscoguttatus × ♂ E. lanceolatu). Hybrid grouper (initial weight 6.76 ± 0.34 g) were fed isonitrogenous and isolipidic diets (16%) with various inositol levels of 0.17 g/kg (J1, the control group), 0.62 g/kg (J2), 1.03 g/kg (J3), 1.78 g/kg (J4), 3.43 g/kg (J5), 6.59 g/kg (J6), respectively. The growth experiment lasted for 8 weeks. The results indicated that dietary inositol had a significant promoting effect on final mean body weight of the J5 and J6 groups and specific growth rate (SGR) of the J3, J4, J5 and J6 groups (P < 0.05). In the serum, superoxide dismutase (SOD) of the J4 group became significantly active compared with that of the control group (P < 0.05), while aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase (AKP) activities in the inositol-treated groups showed distinctly decreased compared with those of the control group (P < 0.05). In the liver, dietary inositol could significantly increase the activities of SOD, catalase (CAT), lysozyme (LYZ) and the contents of total antioxidative capacity (T-AOC) and immunoglobulin M (IgM) (P < 0.05), and distinctly reduce the content of malondialdehyde (MDA) as well as reactive oxygen species (ROS) (P < 0.05). Compared with the control group, the damaged histological morphology of the liver was relieved and even returned to normal after an inositol increase (0.4-3.2 g/kg). In the liver, the remarkable up-regulation of SOD, CAT, glutathione peroxidase (GPX), heat shock protein70 (HSP70) and heat shock protein90 (HSP90) expression levels were stimulated by supply of inositol, while interleukin 6 (IL6), interleukin 8 (IL8) and transforming growth factor β (TGF-β) expression levels were down-regulated by supply of inositol. In head kidney, the mRNA of toll-like receptor 22 (TLR22), myeloid differentiation factor 88 (MyD88) and interleukin 1β (IL1β) expression levels were significantly down-regulated (P < 0.05), which could further lead to remarkable down-regulation of IL6 and tumor necrosis factor α (TNF-α) expression (P < 0.05). These results indicated that high-lipid diets with supply of inositol promoted growth, increased the antioxidant capacity, and suppressed the inflammation of the liver and head kidney by inhibiting the expression of pro-inflammation factors (IL6, IL8, TGF-β and TNF-α). In conclusion, these results indicated that dietary inositol promoted growth, improved antioxidant capacity and immunity of hybrid grouper fed high-lipid diets. Based on SGR, broken-line regression analysis showed that 1.66 g/kg inositol supply was recommended in high-lipid diets of juvenile grouper.