Neutralization of IL ‐ 17A Ameliorates PM2.5‐Induced Cardiac Inflammation and Fibrosis in Rats by Suppressing the TXNIP / NLRP3 Signaling Pathway

Figures

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  FIGURE 1. Expression of IL‐17A and the downstream signaling molecule TRAF6 in cardiac tissue. (A, B) Flow cytometric analysis of splenic lymphocytes; (C–E) IHC images and quantification of the positive area for cardiac IL‐17A and TRAF6; (F–H) Western blot analysis of cardiac IL‐17A and TRAF6. Data are presented as the mean ± SD (n = 6). **p < 0.01.

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  FIGURE 2. Effects of IL‐17A neutralization on cardiac pathology in PM2.5‐exposed rats. (A) Timeline for PM2.5 exposure and IL‐17A antibody treatment. (B) Representative cardiac sections stained with H&E and Masson's trichrome. (C) Cardiac pathological injury score. (D) Fibrotic area. Data are presented as the mean ± SD (n = 6), **p < 0.01.

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  FIGURE 3. Changes in fibrosis‐related markers in cardiac tissue. (A–D) IHC images and quantification of α‐SMA, Collagen I, and Collagen III. (E–H) Western blot analysis of α‐SMA, Collagen I, and Collagen III. (I–K) Expression of mRNA encoding α‐SMA, Collagen I, and Collagen III. Data are presented as the mean ± SD (n = 6), **p < 0.01.

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  FIGURE 4. Cardiac function and injury markers in rats. (A) Representative cardiac echocardiograms. (B–F) LVEF, LVFS, LVEDD, LVESD, and LV mass/BW. (G–H) Serum concentrations of AST and LDH. Data are presented as the mean ± SD (n = 6), *p < 0.05, **p < 0.01.

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  FIGURE 5. Changes in inflammation‐ and oxidative stress‐related indicators in cardiac tissue. (A–D) Western blot analysis of p‐NF‐κB, IL‐6, and IL‐1β. (E, F) ELISA measurements of IL‐6 and IL‐1β. (G–J) Activities of ROS, SOD, GSH, and MDA. (K–M) Western blot analysis of Nrf‐2 and SOD1. Data are presented as the mean ± SD (n = 6), *p < 0.05, **p < 0.01.

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  FIGURE 6. TXNIP expression in rat cardiac tissue. (A, B) IHC images and quantification. (C, D) Western blots and quantification. (E) TXNIP mRNA expression levels. Data are presented as the mean ± SD (n = 6), **p < 0.01.

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  FIGURE 7. Changes in cardiac indicators related to the NLRP3 inflammasome. (A–F) Western blot analysis of NLRP3, ASC, cleaved Caspase‐1 p20, cleaved GSDMD‐N, and IL‐18 in cardiac tissue. Data are presented as the mean ± SD (n = 6). **p < 0.01.

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  FIGURE 8. IL‐17A promotes cardiac fibrosis through the TXNIP/NLRP3 signaling pathway. NRCFs were treated with IL‐17A (100 ng/mL) for 24 h in vitro. (A, B) The protein levels of α‐SMA, Collagen III, IL‐6, and IL‐1β in the control and IL‐17A‐treated groups were measured by Western blot. (C, D) The mRNA and protein levels of TXNIP and NLRP3 in the two groups were quantified by RT‐qPCR and Western blot, respectively. In separate experiments, NRCFs were transfected with siNC or siTXNIP for 48 h, followed by treatment with IL‐17A for 24 h. (E, F) mRNA and protein expression of TXNIP and NLRP3 in the siNC and siTXNIP groups were assessed; (G, H) protein levels of α‐SMA, Collagen III, IL‐6, and IL‐1β in these two groups were measured as well. Data are presented as the mean ± SD (n = 4), *p < 0.05.