Inhibition of CX3C receptor 1-mediated autophagy in macrophages alleviates pulmonary fibrosis in hyperoxic lung injury

Abstract

AimsTo investigate the role of CX3CR1 in hyperoxic lung injury induced pulmonary fibrosis. Materials and methodsHyperoxic lung injured mice were used as the disease model. Pulmonary fibrosis was determined by H&E and Masson's staining. Autophagy was investigated by western blot, immunofluorescence staining, and transmission electron microscopy. Key findingsWe observed that increased CX3CR1 expression corresponded with increased pulmonary fibrosis. Additionally, silencing of CX3CR1 significantly alleviated the fibrosis when compared to the control. We observed that exposure of mouse to hyperoxic environment increased macrophage levels along with an increased CD11b expression in the lung tissues. Subsequently, we also observed an increased expression of LC3-II and decreased p62 expression in hyperoxic mice models, suggesting the potential role of hyperoxia induced autophagy. CD11b and LC3/CX3CR1 were expressed and co-localized in a manner indicating CX3CR1 indeed does regulate macrophage autophagy in the hyperoxic lung injury model. We observed a decrease in hyperoxia-associated fibrosis, along with a decrease in autophagy when we used 3-MA (autophagy inhibitor) in our hyperoxic lung injury model. To elucidate the pathway through which CX3CR1 regulated autophagy, we further analyzed the Akt1 pathway. Our experimental results indicated that the Akt1 inhibitor (A-674563) did significantly decrease macrophage autophagy and fibrosis in hyperoxic mice models. SignificanceThus, our data indicates a novel role of CX3CR1 in regulation of macrophage autophagy and promotion of pulmonary fibrosis in hyperoxic lung injured mice.