Clara cell secretory protein 16 protects against PM2.5-induced ferroptosis in mouse lung epithelial cells in a concentration-dependent manner
Year: 2025
Type: article
Abstract: Background Clara cell secretory protein 16 (CC16) has been reported to exert anti-inflammatory and antioxidant 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 Fe 2+ 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 PM2.5induced respiratory disease.
Author: Aili Wang
Institution: Wuhan No.1 Hospital
Language: English
Subfield: Pulmonary and Respiratory Medicine
Field: Medicine
Domain: Health Sciences
Open Access status: bronze
OpenAlex ID: W4415725908