Dang Gui Bu Xue Tang, a conventional Chinese herb decoction, ameliorates radiation-induced heart disease via Nrf2/HMGB1 pathway

Figures

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  FIGURE 1. The protective effect of DBT on the radiation-induced abnormal weight loss. (A) The representative images of body weighing at 8 weeks after radiation; (B) Trends in body weight change. *p < 0.05 of Radiation versus Control; † p < 0.05 of RDBT51 versus Radiation; # p < 0.05 of RDBT11 versus Radiation. DBT, Dang Gui Bu Xue Tang; RAM, Radix Astragali membranaceus; RAS, Radix Angelicae sinensis; RDBT51, Radiation with DBT (RAM: RAS = 5:1); RDBT11, Radiation with DBT (RAM: RAS = 1:1); RDBT15, Radiation with DBT (RAM: RAS = 1:5); The “R” in the “RDBT” stands for Radiation, while the two digits following RDBT represent the ratio between RAM and RAS, the same below.

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  FIGURE 2. The DBT ameliorates the inflammation- and myocardial injury-related biomarkers in the serum of mice at 4 and 8 weeks after radiation. (A) TNF-α; (B) IL-6; (C) cTnT; (D) CK-MB. *p < 0.05 versus Control; † p < 0.05 versus Radiation. DBT, Dang Gui Bu Xue Tang; RAM, Radix Astragali membranaceus; RAS, Radix Angelicae sinensis; RDBT51, Radiation with DBT (RAM: RAS = 5:1); RDBT11, Radiation with DBT (RAM: RAS = 1:1); RDBT15, Radiation with DBT (RAM: RAS = 1:5).

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  FIGURE 3. The DBT improves radiation-induced left ventricular systolic dysfunction. (A) Representative echocardiograms obtained at 4 and 8 weeks after radiation treatment; (B) Quantitative statistical analyses of FS and EF. The other concerning quantitative statistical analyses of cardiac function related parameters are summarized in Table 1. DBT, Dang Gui Bu Xue Tang; EF, ejection fraction; FS, fractional shortening; RAM, Radix Astragali membranaceus; RAS, Radix Angelicae sinensis; RDBT51, Radiation with DBT (RAM: RAS = 5:1); RDBT11, Radiation with DBT (RAM: RAS = 1:1); RDBT15, Radiation with DBT (RAM: RAS = 1:5).

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  FIGURE 4. The DBT alleviates radiation-induced cardiac fibrosis. (A) Representative microphotographs of Sirius red staining obtained at 8 weeks after radiation; (B) Quantitative results of collagen deposition percentage; (C), (D), and (E) The myocardial mRNA levels of TGF-β, Collagen-I and Collagen-III relative to GAPDH were analysed by real-time qPCR. *p < 0.05 versus Control; † p < 0.05 versus Radiation. DBT, Dang Gui Bu Xue Tang; RAM, Radix Astragali membranaceus; RAS, Radix Angelicae sinensis; RDBT51, Radiation with DBT (RAM: RAS = 5:1); RDBT11, Radiation with DBT (RAM: RAS = 1:1).

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  FIGURE 5. The DBT reduces the fibrosis-related biomarkers in the neonatal mouse myocardial fibroblast culture supernatant at 24 and 48 h after radiation. (A) Galectin-3; (B) HMGB1. *p < 0.05 versus Control; † p < 0.05 versus Radiation. DBT, Dang Gui Bu Xue Tang; HMGB1, high mobility group box 1; RAM, Radix Astragali membranaceus; RAS, Radix Angelicae sinensis; RDBT51, Radiation with DBT (RAM: RAS = 5:1); RDBT11, Radiation with DBT (RAM: RAS = 1:1); RDBT15, Radiation with DBT (RAM: RAS = 1:5).

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  FIGURE 6. The DBT reduces the fibrosis-related biomarkers in the serum of mice at 4 and 8 weeks after radiation. (A) Galectin-3; (B) HMGB1. *p < 0.05 versus Control; † p < 0.05 versus Radiation. DBT, Dang Gui Bu Xue Tang; HMGB1, high mobility group box 1; RAM, Radix Astragali membranaceus; RAS, Radix Angelicae sinensis; RDBT51, Radiation with DBT (RAM: RAS = 5:1); RDBT11, Radiation with DBT (RAM: RAS = 1:1); RDBT15, Radiation with DBT (RAM: RAS = 1:5).

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  FIGURE 7. The regulatory effect of radiation and DBT on mouse myocardial Nrf2 and HMGB1 protein levels. (A) Representative western blot bands; (B) Quantitative results of Western blot. *p < 0.05 versus Control; † p < 0.05 versus Radiation. DBT, Dang Gui Bu Xue Tang; HMGB1, high mobility group box 1; RAM, Radix Astragali membranaceus; RAS, Radix Angelicae sinensis; RDBT51, Radiation with DBT (RAM: RAS = 5:1); RDBT11, Radiation with DBT (RAM: RAS = 1:1).

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  FIGURE 8. Inhibition of Nrf2 compromises the protective effect of DBT on radiation-induced deleterious changes in myocardial fibroblasts. After administration of radiation, myocardial fibroblasts were immediately changed to fresh normal medium, DBT51 medium (1 mg/ml), or DBT51 medium with 1 μM ML385 (specific inhibitor of Nrf2) for 48 h and then collected for analyses. (A) Representative western blot bands; (B) Quantitative results of Western blot; (C), (D), and (E) Quantitative results of real-time qPCR for TGF-β, Collagen-I and Collagen-III. *p < 0.05 versus Control; † p < 0.05 versus Radiation; # p < 0.05 versus DBT51. DBT, Dang Gui Bu Xue Tang; HMGB1, high mobility group box 1; RAM, Radix Astragali membranaceus; RAS, Radix Angelicae sinensis; RDBT51, Radiation with DBT (RAM: RAS = 5:1).