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Diabetes mellitus is one of the most common endocrine metabolic disorder that has caused significant Morbidity and Mortality due to the micro and macro vascular vascular complications. In current study, NPLME (Nymphaea pubescent leaf methanolic extract) is beneficial in controlling the blood glucose levels, improved the lipid metabolism and also prevented the diabetic complications in experimental diabetic rats. We have analyzed the effect of Nymphaea Pubescens on serum glucose, triglycerides, cholesterol, HDL-Cholesterol, LDL-Cholesterol, ALT, AST, glycosylated hemoglobin, liver glycogen, in vivo antioxidant parameters (catalase, lipid peroxidation (MDA) and reduced glutathione) in experimentally induced diabetic rats. Furthermore, Guggultetrol of Nymphaea pubescent leaves exhibited a binding interactions with the enzyme glucokinase in the in-silico analysis. Our studies revealed that NPLME showed a significant antidiabetic activity in diabetic rats compared to healthy controls and it supports the traditional use of Nymphaea pubescent leaf by the Ayurvedic physicians for the control of diabetes.