Causes of diabetes could be associated with many dysfunctions regarding the regulation of insulin and progression of pancreatic beta cells. Phytonanotherapy provides a symbiotic properties of metal nanoparticles and plants as they exhibit medicinal benefits which may be biologically equivalent for synthetic drugs. The present study focused on rapid and ecofriendly biosynthesis of gold nanoparticles using Eclipta alba (EA-AuNPs) and evaluated its pharmacological efficacy against pathogenic bacteria and streptozotocin (STZ) induced apoptosis in RIN-5F cell line. The bioreduction of auric chloride was attained through capping or stabilizing molecules offered by the phytoconstituents present in Eclipta alba. The green synthesized EA-AuNPs was characterized by spectrometric studies, such as UV–vis spectroscopy, X-ray diffraction analysis (XRD), Fourier transform infrared spectroscopic (FTIR) study, dynamic light scattering (DLS), and by microscopic studies such as high-resolution transmission electron microscopy (TEM), scanning electron microscopy (SEM) and atomic force microscopy (AFM). The synthesized EA-AuNPs showed surface plasmon resonance band at 536 nm and the particle was spherical about 26 nm with a good zeta potential value of −12mv. The anti-apoptotic potential of EA-AuNPs was proved by evaluating the NF-κB mediated apoptotic pathway in STZ-induced diabetes in RIN-5F cell line. We further showed the free radical scavenging potential of EA-AuNPs and stabilized EA-AuNPs demonstrated strong antibacterial activity against human pathogenic bacteria. Therefore, we conclude that Eclipta alba has a good potential to synthesize AuNPs and the synthesized EA-AuNPs are promising for biomedical applications in the field of nanomedicine. © 2020 Elsevier B.V.