Peperomia pellucida (L.) is commonly used as a herbal plant. Its effectiveness in treating inflammatory diseases, digestive disorders, and cancer in tropical and subtropical countries was introduced, especially in field of folk medicine. However, this plant species has not been studied widely in Vietnam, especially for its biological activities. This study was done to determine the antioxidant capacity of P. pellucida by using in vitro and in vivo methods, as well as its inhibitory ability to α-amylase enzyme activity. The total polyphenolic and flavonoid contents of P. pellucida extract were reported to be 359.91±0.77 mg GAE/g and 200.28±1.23 mg QE/g extract, respectively. The results showed the in vitro antioxidant activity of P. pellucida extract in four methods, including DPPH, and ABTS.+, RP and TAC, had EC50 values of 730.34 µg/mL, 84.33 µg/mL, 95.28 µg/mL, respectively, and Abs0.5 value of 114.73 µg/mL. Under H2O2-induced oxidative stress, fruit flies that were raised in the feed medium supplemented with a concentration of 1 mg/mL of P. pellucida extract showed their average survival time, 50% survival time, and 10% survival time at 1.6 times, 1.8 times, and 1.62 times, respectively, higher than those of the control treatment. The ability to inhibit the α-amylase activity in P. pellucida extract was determined with an EC50 value of 115.32±2.65 µg/mL compared with the commercial drug of 18.67±0, 01 µg/mL. The research results showed that P. pellucida is a potential species in the study of natural compounds with antioxidant and antidiabetic activities.