Objective: A C2C12 myotube cell insulin resistance (C2C12-IR) model was established to explore the effect of isothiocyanate extracted from Moringa oleifera (MIC-1) on C2C12 myotube cell insulin resistance. Methods: Palmitic acid (PA) was used to induce C2C12 myotube cells to establish a stable C2C12-IR model， and the cells were treated with different concentrations of MIC-1 (0， 2， 4， 8， 16 μmol/L) for 16 h. The cell viability of C2C12-IR cells was detected by MTT assay， and the effect of MIC-1 on glucose metabolism of C2C12-IR cells was observed. Cellular oxidative damage [nitric oxide (NO)， malondialdehyde (MDA) and glutathione peroxidase (GSH)] were tested. The expression levels of related proteins in the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway were detected by using Western blot assay. Results: The optimal modeling conditions for the C2C12-IR model were determined， that is， C2C12 myotube cells were treated with 0.5 mmol/L PA for 16 h. Compared with the C2C12-IR group， MIC-1 significantly increased the glucose consumption and glycogen content of C2C12-IR cells in a dose-dependent manner. MIC-1 significantly decreased the level of MDA and NO， and significantly increased the level of GSH in C2C12-IR cells. In addition， MIC-1 significantly increased the expression levels of phosphoinositide-dependent protein kinase-1 (PDK1)， p-AKT， and glucose transporter 4 (GLUT4). Conclusion: MIC-1 improves insulin resistance by inhibiting oxidative stress.