Ultrafiltration Extract of Radix Angelica Sinensis and Radix Hedysari Attenuates Risk of Low-Dose X-Ray Radiation-Induced Myocardial Fibrosis In Vitro

Figures

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  Figure 1. Identification of CFs and RAS-RH inhibited the proliferation of cardiac fibroblasts after X-ray radiation. (a, b) Representative images of identification of CFs by immunofluorescence staining of α-SMA actin and Vimentin were used to identify the purity of myocardial fibroblasts isolated by double enzyme digestion. Nuclei were stained blue; α-SMA and Vimentin were stained red, magnification ×200. (c) Representative results of shape of CFs by fluorescence microscope in the control group and X-ray group (at 48h after irradiation). Magnification ×40. (d) RAS-RH inhibits proliferation rate of primary CFs after X-ray radiation depended on the manner of dose-time. CFs were treated with 3, 6, and 9 μg/ml of RAS-RH after X-ray irradiation for the indicated times. Proliferation rate of CFs was determined by CCK-8 assay (n = 5, ∗P < 0.05, ∗∗P < 0.01 vs. the control group, #P < 0.05, ##P < 0.01 vs. X-ray group). (e) TGF-β1-positive cells in each group with immunofluorescence staining. Nuclei were stained blue and TGF-β1 stained red, magnification ×200.

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  Figure 2. RAS-RH increased the apoptosis of CFs after X-irradiation. (a) CFs were stained with annexin V-FITC and propidium iodide. The percentages of apoptosis were detected by flow cytometry. (b) Histogram that represents a statistical analysis of image a (n = 3, ∗P < 0.05 vs. the control group, #P < 0.05 vs. X-ray group, #P < 0.05, ##P < 0.01 vs. X-ray group).

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  Figure 3. Oxidative stress from X-irradiation was attenuated by RAS-RH. ROS were analyzed by flow cytometry in the Con, X-ray, and RAS-RH + X-ray groups (n = 3, ∗P < 0.05 vs. the control group, #P < 0.05 vs. X-ray group).

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  Figure 4. RAS-RH alleviated the expression of TGF-β1, Col1, and α-SMA in CFs after X-ray radiation. (a, b) Representative images indicate the protein level of TGF-β1, COL1, and α-SMA were analyzed with western blotting by Image-J in the Con, X-ray, and RAS-RH + X-ray groups (n = 3, ∗P < 0.05, ∗∗P < 0.01 vs. the control group, #P < 0.05, ##P < 0.01 vs. X-ray group). (c) Quantitative real-time RT-PCR analysis of TGF-β1, COL1, and α-SMA mRNA levels in the Con, X-ray, and RAS-RH + X-ray groups (n = 3, ∗P < 0.05, ∗∗P < 0.01 vs. the control group, #P < 0.05, ##P < 0.01 vs. X-ray group).