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The effect of Lactobacillus plantarum- fermentation on the anti-diabetic functionality of Momordica charantia was examined using a high-fat-diet and low-dose streptozocin-induced type 2 diabetic rat model. Fermented Momordica charantia juice (FMCJ) administration mitigated the hyperglycemia, hyperinsulinemia, hyperlipidemia, and oxidative stress in diabetic rats more favorably than the non-fermented counterpart. Treatments with FMCJ improved ergosterols and lysomonomethyl-phosphatidylethanolamines metabolisms more effectively. Supplement of FMCJ regulated the composition of the gut microbiota, such as increased the abundance of Bacteroides caecigallinarum , Oscillibacter ruminantium, Bacteroides thetaiotaomicron, Prevotella loescheii , Prevotella oralis , and Prevotella melaninogenica , in diabetic rats compared with untreated diabetic rats. Moreover, FMCJ-treated diabetic rats exhibited higher concentrations of acetic acid, propionic acid, butyric acid, total short-chain fatty acids and lower pH values in colonic contents than that in non-fermented juice-treated rats. These results demonstrated that Lactobacillus plantarum -fermentation enhanced the anti-diabetic property of MC juice by favoring the regulation of gut microbiota and the production of SCFAs. Unlabelled Image • Fermentation improved serum glucose, insulin, and lipids in MCJ-treated diabetes. • Fermentation enhanced the regulatory effect of MCJ on lipid metabolism in diabetes. • Fermentation regulated the composition of gut microbiota in MCJ-treated diabetes. • Fermentation elevated colonic SCFAs contents in MCJ-treated diabetes. [ABSTRACT FROM AUTHOR]