Objective: The systematic study of effective alternative anti-diabetic drugs has great importance to manage diabetes as well as other oxidative stress-related diseases. According to previous research, root and bark of Mussaenda macrophylla plant has anti-microbial, anti-coagulant, anti-inflammatory, and hepatoprotective activity. Ethnomedicinal data shows that Mussaenda macrophylla is used to treat diabetes as well as oxidative stress. The objective of this research is to investigate in vitro anti-diabetic and anti-oxidant activity of root extract of Mussaenda macrophylla. Methods: DPPH free radical scavenging assay was used to detect anti-oxidant potency of ethanol and methanol root extract of the plant and expressed as % of radicle inhibition. Anti-diabetic activity was determined by the glucose diffusion method using a glucose oxidase kit and results were expressed as mean±SD. Results: The ethanol root extract at the concentration of 50 mg/ml and 100 mg/ml showed better glucose diffusion inhibition than that of methanol extract at the same concentration on increasing time interval. Ethanol extract at the concentration 100 µg/ml displayed better DPPH scavenging activity (89.83±0.19 %) than that of methanol extract (86.61±0.75%). Conclusion: This study concluded that ethanol and methanol root extract of Mussenda macrophylla have potent anti-diabetic as well as anti-oxidant activity but further advance research is necessary in the animal model.