Pirfenidone inhibits the polarization of M2 macrophages by down-regulating IRF4 to reduce the occurrence and development of radiation-induced lung fibrosis
Fang Min1, Ying Hangjie1, Hang Qingqing2, Chen Yamei1, Chen Ming1
1Department of Radiation Oncology, The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Insititute of Basic Medicine and Cancer (IBMC),Chinese Academy of Sciences,Hangzhou, 310022, China; 2The Second Clinical Medical College of Zhejiang Chinese Medical University,Hangzhou 310053, China
Abstract：Objective To explore the preventive and therapeutic effect of pirfenidone (PFD) on radiation-induced lung fibrosis (RILF) and its mechanism. Methods 40 female C57/BL6 mice were randomly divided into 4 groups:negative control group (NC), PFD treatment group (PFD), radiation treatment group (RT) and radiation plus PFD treatment group (RT+PFD). Mice in RT and RT+PFD groups received a single whole lung X-ray consisting of a 50Gy dose of radiation, delivered by small animal radiation research platform (SARRP). PFD at a dose of 300mg/kg was administered orally 2h before irradiation for 150d. HE and Masson staining were used to detect the infiltration of inflammatory cells and the degree of pulmonary fibrosis. Quantitative real-time PCR (qPCR) and Western blotting (WB) were adopted to detect the expression levels of M1/M2 macrophage phenotypic markers. The expression levels of arginase-1(ARG-1), chitinase 3-like protein 3(YM-1) and interferon regulatory factor-4(IRF4) of macrophages stimulated with IL-4 and IL-13 were detected by WB. In addition, immunofluorescence staining was used to detect the expression and translocation of IRF4 in macrophages among different treatment groups. Results HE and Masson staining showed that PFD could significantly inhibit radiation-induced infiltration of inflammatory cells and fibrosis in lung tissues. The M2 macrophages and expression levels of ARG-1 and YM-1 were down-regulated in the RT+PFD group. Cell experiments further confirmed that PFD could significantly inhibit the polarization of macrophages to M2 induced by IL-4+IL-13, which was mainly related to the down-regulation of IRF4. Conclusion PFD has a preventive and therapeutic effect on RILF by inhibiting IRF4 and reducing the polarization of macrophages to M2.
Fang Min,Ying Hangjie,Hang Qingqing et al. Pirfenidone inhibits the polarization of M2 macrophages by down-regulating IRF4 to reduce the occurrence and development of radiation-induced lung fibrosis[J]. Chinese Journal of Radiation Oncology, 2022, 31(6): 562-568.
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