People of all ages are affected by Diabetes Mellitus (DM), a metabolic illness that is very prevalent. The development of the illness is significantly influenced by environmental variables and changes in lifestyle. The anti-diabetic activity of ethanolic crude extract of Luffa acutangula and its active fractions were evaluated by in-vitro assays. Extraction of Luffa acutangula leaves was carried out using ethanol and further, Ethyl-acetate and n-Butanol fractions were obtained. To evaluate the potential hypoglycemic effects of ethanolic extract of Luffa acutangula (EELA), Ethyl acetate fraction of Luffa acutangula (EALA), n-butanol fraction of Luffa acutangula (NBLA) and Acarbose (standard drug), (100-1000 g/ml) concentrations were used for the - amylase and -glucosidase enzyme inhibitory study. EELA, EALA, NBLA & Metronidazole were also studied for glucose transport across yeast cells at 5mM, 10mM, and 25mM concentrations of glucose. From the above studies carried out, the active fraction was subjected for the MIN-6 cell line study. The insulin secretagogue actions of EELA, EALA & NBLA were investigated utilizing a mouse insulinoma beta cell line (MIN6-). The study has revealed that the EALA shows the highest anti-diabetic potential in comparison to the EELA & NBLA.