Arsenic (As) polluted food chain has become a serious issue for the growth and development of humans, animals and plants. Nitric oxide (NO) or silicon (Si) may mitigate As toxicity. However, the combined application of NO and Si in mitigating As uptake and phytotoxicity in Brassica juncea is unknown. Hence, the collegial effect of sodium nitroprusside (SNP), a NO donor and Si application on B. juncea growth, gas exchange parameters, antioxidant system and As uptake was examined in a greenhouse experiment. Arsenic toxicity injured cell membrane as signposted by the elevated level of malondialdehyde (MDA) and hydrogen peroxide (HO), thus decreasing the growth of stressed plants. Moreover, As stress negatively affected gas exchange parameters and antioxidative system of plants. However, NO or/and Si alleviated As induced oxidative stress through increasing the activity of superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR), glutathione S-transferase (GST), glutathione (GSH), along with thiol and proline synthesis. Furthermore, plants treated with co-application of NO and Si showed improved growth, gas attributes and decreased As uptake under As regimes. The current study highlights that NO and Si synergistically interact to mitigate detrimental effects of As stress through reducing As uptake. Our findings recommend combined NO and Si application in As spiked soils for improvement of plant growth and stress alleviation.