Abstract
Background Clara cell secretory protein 16 (CC16) has been reported to exert anti-inflammatory and anti-oxidant effects. However, its underlying mechanism remains unclear. This study aimed to investigate the protective effect and mechanism of CC16 using an in vitro model of PM2.5-induced mouse pulmonary epithelial cells (TC-1), with a specific focus on its concentration-dependent effects.
Methods:
TC-1 cells were exposed to PM2.5 to induce inflammatory injury and ferroptosis, followed by treatment with CC16 at different concentrations (0.25, 0.5, and 1.0 µg/mL). TC-1 cells were divided into six groups: control, CC16, PM2.5, PM2.5 + CC16 (0.25 μg/mL), PM2.5 + CC16 (0.5 μg/mL), and PM2.5 + CC16 (1.0 μg/mL) groups. Cell viability was assessed using the Cell Counting Kit-8 assay. Levels of inflammatory cytokines (interleukin (IL)-5, IL-6, IL-13, IL-17A, and IL-1β) in the supernatant were measured by enzyme-linked immunosorbent assay. Protein expression levels of Nrf2, ACSL4, SLC7A11, and GPX4 were detected using western blotting. Intracellular reactive oxygen species (ROS) were detected with a fluorescent probe, and mitochondrial membrane potential (MMP) was measured by JC-1 staining. Glutathione (GSH), malondialdehyde (MDA), and Fe2+ content were measured using relevant kits. Mitochondrial ferroptosis features were observed by transmission electron microscopy (TEM).
Results:
PM2.5 exposure significantly reduced cell viability, aggravated inflammation, induced ferroptosis, and disrupted the integrity of mouse pulmonary epithelial cells. CC16 treatment reversed these effects in a concentration-dependent manner.
Conclusions:
CC16 effectively mitigates PM2.5-induced cellular injury in mouse pulmonary epithelial cells through inhibition of ferroptosis, with its protective effect showing a clear concentration dependence. These findings suggest CC16 as a novel strategy for PM2.5-induced respiratory disease.
Keywords:
CC16; PM2.5; ferroptosis; pulmonary epithelial cells.