Urban particulate matter (UPM), an air pollutant-absorbing toxic substance, can access alveoli, leading to pulmonary diseases. Studies have shown that the water-soluble components of UPM (WS-UPM), containing main toxic substances, can induce oxidative damage in lung cells. In this study, the UPM particle size and composition were detected via instrumental analysis. The isoflavones (biochanin A (BCA), formononetin and daidzein) from chickpeas possess biological antioxidant properties. The present study aimed to investigate the mechanism of the oxidative damage induced by WS-UPM, and the protective role of isoflavones in human alveolar basal epithelial cells. The antioxidant activity of BCA, formononetin and daidzein was investigated through the total reduction capacity, diphenylpicrylhydrazine radical (DPPH), superoxide radical, and hydroxyl radical scavenging capacity detection. We also established cell models in vitro to further explore the BCA-protective mechanism. BCA presented a significant protection, and increased the levels of antioxidant makers including superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH). The effects were also reflected as the reduction of malondialdehyde (MDA) and nitric oxide (NO). Moreover, results obtained from RT-PCR and western blot techniques revealed that MEK5/ERK5 played an indispensable role in regulating the antioxidant effect of BCA, alleviating WS-UPM-induced lung injury. Furthermore, BCA mitigated WS-UPM-exposed damage through upregulating the Nrf2 signaling pathway to enhance the antioxidase expression downstream of Nrf2. In summary, our findings indicated that the WS-UPM-induced pulmonary disease was involved in oxidative stress and the MEK5/ERK5-Nrf2 signaling pathway, and BCA regulated the WS-UPM-induced lung damage via upregulation of the MEK5/ERK5-Nrf2 pathway.