Journal: Journal of Cellular and Molecular Medicine

Article Title: Pirfenidone modulates macrophage polarization and ameliorates radiation‐induced lung fibrosis by inhibiting the TGF‐β1/Smad3 pathway

doi: 10.1111/jcmm.16821

Figure Lengend Snippet: Pirfenidone attenuates pulmonary fibrosis induced by whole‐lung radiation. A, Photographs of mice in the four groups after 84 days of treatment. B, Photographs of the lung tissue in the four groups. C, The mean left lung weights of mice in the four groups ( n = 5 per group). The values are the means ± SD, * p < 0.05, ** p < 0.01

Article Snippet: Pirfenidone (PFD) is a multipotent pyridone analog that was discovered by the MARNAC Company in 1974.

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: Pirfenidone modulates macrophage polarization and ameliorates radiation‐induced lung fibrosis by inhibiting the TGF‐β1/Smad3 pathway

doi: 10.1111/jcmm.16821

Figure Lengend Snippet: Pirfenidone effectively inhibited alveolar inflammation, pulmonary fibrosis and collagen deposition induced by lung irradiation. A, HE and Masson staining of lung tissues in the four groups and semiquantitative analysis of the degree of pulmonary fibrosis. The values are the means ± SD, ** p < 0.01, *** p < 0.001. B, IHC staining of collagen I and collagen IV in lung tissue from the four groups and semiquantitative analysis of collagen I and collagen IV expression. Scale bar = 100 μm. The values are the means ± SD, ** p < 0.01, *** p < 0.001

Article Snippet: Pirfenidone (PFD) is a multipotent pyridone analog that was discovered by the MARNAC Company in 1974.

Techniques: Irradiation, Staining, Immunohistochemistry, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Pirfenidone modulates macrophage polarization and ameliorates radiation‐induced lung fibrosis by inhibiting the TGF‐β1/Smad3 pathway

doi: 10.1111/jcmm.16821

Figure Lengend Snippet: Pirfenidone inhibits ionizing radiation‐induced M2 polarization. A, The expression of CD68, CD79 and CD3 in the lung tissue of mice in the different treatment groups was determined by IHC analysis. Scale bar = 100 μm. B, qRT‐PCR was used to measure the expression of ARG‐1, YM‐1, iNOS and TNF‐α in the lung tissue of mice in the different treatment groups. ( n = 3 per group). The values are the means ± SD, * p < 0.05, C, Western blotting was used to measure the expression of ARG‐1 and YM‐1 in the lung tissue of mice in the different treatment groups. Data from individual animals ( n = 3 per group). The values are the means ± SD, * p < 0.05, ** p < 0.01

Article Snippet: Pirfenidone (PFD) is a multipotent pyridone analog that was discovered by the MARNAC Company in 1974.

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Pirfenidone modulates macrophage polarization and ameliorates radiation‐induced lung fibrosis by inhibiting the TGF‐β1/Smad3 pathway

doi: 10.1111/jcmm.16821

Figure Lengend Snippet: Pirfenidone inhibits the polarization of M2 macrophages by downregulating NF‐κB p50 expression. A, Cytotoxic effects of PFD on RAW264.7 cells and BMDMs. Plots showing the effects of different concentrations of PFD on the growth of RAW264.7 cells and BMDMs during the indicated times. The values are the means ± SD, * p < 0.05, ** p < 0.01, when comparing 1000 μg/ml with 0 μg/ml. B, qRT‐PCR was used to analyse the quantitative mRNA expression of the M2 phenotypic markers ARG‐1 and YM‐1 in the 4 different treatment groups of RAW264.7 cells and BMDMs. The values are the means ± SD, ## p < 0.01 and ### p < 0.001, when compared with the untreated group, * p < 0.05, ** p < 0.01, *** p < 0.001, when compared with the IL‐4 + IL‐13–stimulated group. C, Western blots and associated densitometry analysis of the protein expression of the M2 phenotypic markers ARG‐1, YM‐1 and NF‐κB p50 in the 4 different treatment groups of RAW264.7 cells and BMDMs. The values are the means ± SD, ## p < 0.01 and ### p < 0.001, compared with the untreated group, * p < 0.05, ** p < 0.01, *** p < 0.001 compared with the IL‐4 + IL‐13–stimulated group. The tests were repeated in three independent experiments

Article Snippet: Pirfenidone (PFD) is a multipotent pyridone analog that was discovered by the MARNAC Company in 1974.

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Pirfenidone modulates macrophage polarization and ameliorates radiation‐induced lung fibrosis by inhibiting the TGF‐β1/Smad3 pathway

doi: 10.1111/jcmm.16821

Figure Lengend Snippet: Pirfenidone inhibits irradiation‐induced activation of the TGF‐β1/Smad3 signalling pathway in vivo and in vitro. A, The expression and quantitative analysis of TGF‐β1, p‐Smad3, Smad3 and β‐actin in lung tissue from the four different treatment groups. Data from individual animals ( n = 3 per group); the values are the means ± SD, * p < 0.05, ** p < 0.01. In Figures and , we run the same four subgroups, NC, PFD, RT, RT + PFD and stain YM‐1, ARG‐1 (in Figure ) and TGF‐β1, p‐Smad3, Smad3 (in Figure ). Therefore, the β‐actin in Figure and Figure is the same band. B, Cytotoxic effects of PFD on the proliferation of MLE‐12 cells and PMVECs. Plots showing the effects of different concentrations of PFD on the growth of MLE‐12 cells and PMVECs over the indicated times. The values are the means ± SD. MLE‐12 cells: *** p < 0.001, 1000 μg/ml compared with 0 μg/ml PMVECs: ** p < 0.05, *** p < 0.001, when comparing 100 μg/ml with 0 μg/ml; ### p < 0.001, when comparing 1000 μg/ml with 0 μg/ml. C, The expression and quantitative analysis of TGF‐β1, p‐Smad3, Smad3 and β‐actin in MLE‐12 cells and PMVECs after RT and PFD treatment. The values are the means ± SD, # p < 0.05, compared with the untreated group, * p < 0.05, ** p < 0.01, *** p < 0.001, compared with the IL‐4 + IL‐13–stimulated group. The tests were repeated in three independent experiments

Article Snippet: Pirfenidone (PFD) is a multipotent pyridone analog that was discovered by the MARNAC Company in 1974.

Techniques: Irradiation, Activation Assay, In Vivo, In Vitro, Expressing, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Pirfenidone modulates macrophage polarization and ameliorates radiation‐induced lung fibrosis by inhibiting the TGF‐β1/Smad3 pathway

doi: 10.1111/jcmm.16821

Figure Lengend Snippet: Pirfenidone is involved in the crosstalk between alveolar epithelial cells and macrophages. A, The protein levels of CCL2 and CXCL1 in MLE‐12 cell culture supernatants were detected by ELISA after irradiation with 4 different doses. The values are the means ± SD, * p < 0.05, ** p < 0.01, compared with 0 Gy. B, The protein expression of TGF‐β1 in culture supernatants of RAW264.7 cells in the groups treated with IL‐4 + IL‐13 and PFD was measured by ELISA. The values are the means ± SD, ** p < 0.01, compared with 0 Gy. C, The migration of RAW264.7 cells was assessed in a Transwell assay. Representative images (left) and statistical analysis (right) show the migration of RAW264.7 cells induced by different conditioned media. Scale bar = 200 μm. The values are the means ± SD, *** p < 0.001. D, The expression levels of p‐Smad3, Smad3 and β‐actin in MLE‐12 cells cultured in conditioned media collected from RAW264.7 cells in the different treatment groups were analysed by Western blotting. The values are the means ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001. The tests were repeated in three independent experiments

Article Snippet: Pirfenidone (PFD) is a multipotent pyridone analog that was discovered by the MARNAC Company in 1974.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Irradiation, Expressing, Migration, Transwell Assay, Western Blot