Blockade of Aquaporin 4 Inhibits Irradiation-Induced Pulmonary Inflammation and Modulates Macrophage Polarization in Mice

To investigate the effects of aquaporin 4 (AQP4) inhibitor in irradiation-induced pulmonary inflammation in mice. A single dose of 75 Gy was delivered to the left lung of mice to induce radiation pneumonitis. For inhibition of AQP4, 200 mg/kg of TGN-020 was administered i.p. one time per 2 days post-irradiation. Blockade of AQP4 with TGN-020 resulted in the inhibition of inflammatory cell infiltration and the downregulation of inflammatory cytokines (IL-6, IL-17, and TGF-β), chemokines (MIP1a and MCP1), fibrosis-related (Col3al and Fn1), and M2 macrophage marker (Arg1) post-irradiation. Immunofluorescence staining indicated that there was significant fewer M2 macrophage infiltration in the irradiated lung tissues from mice treated with TGN-020. Additionally, depletion of macrophages with liposome clodronate resulted in alleviated lung injury induced by irradiation. Furthermore, adoptive transfer of M1 or M2 macrophages into clodronate-treated mice was performed. The results showed that the administration of M2 macrophages fully reversed the clodronate-induced beneficial effect on inflammation score, thickness, and fibrosis. However, transfer of M1 macrophages only impacted the inflammation score and thickness and did not affect lung fibrosis. AQP4 blockade alleviated the development and severity of irradiated lung damage. This was associated with attenuated infiltration of inflammatory cell, decreased production of pro-inflammatory cytokines, and inhibited activation of M2 macrophages.