The involvement of Nox4 in fine particulate matter exposure-induced cardiac injury in mice

Epidemiological studies have confirmed that ambient fine particulate matter (PM_2.5) exposure is associated with cardiovascular disease (CVD). However, the underlying mechanisms in PM_2.5 exposure-induced heart injury are largely unknown. It has been acknowledged that NADPH oxidase (Nox) 4 plays a critical role in CVD development. To investigate the acute effects of PM_2.5 on the mouse heart and the role of Nox4 in PM_2.5 exposure-induced cardiac injury, C57BL/6J mice were instilled with saline or 1.5, 3.0, 6.0 mg/kg BW PM_2.5 suspension for two weeks (five days per week). The levels of malondialdehyde (MDA), super oxide dismutase (SOD), inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α) and interleukin (IL)-1β in heart supernatants were determined using related kits. The expression of Nox4, p67^phox, p47^phox and p22^phox in heart tissue was evaluated by immunofluorescence staining or Western blotting, respectively. Protein levels of p53, Bax, Bcl-2 and Caspase-3 in the heart were examined using immunohistochemical staining and Western blotting. TUNEL assay was used to measure myocardial apoptosis. PM_2.5 exposure leads to obvious cardiac injury. PM_2.5 exposure increases MDA level and iNOS activity, and decreases activity of SOD in heart supernatants of mice. High levels of TNF-α and IL-1β in heart supernatants of mice with PM_2.5 instillation were determined. Nox4 and Nox-associated subunits such as p67^phox, p47^phox and p22^phox expression levels were increased in heart tissue of mice after PM_2.5 exposure. Additionally, PM_2.5 exposure causes myocardial apoptosis in the mouse heart. This study suggested that Nox4 is involved in PM_2.5 exposure-induced cardiac injury in mice.