Intervention of Exosome-Mediated Radiation Bystander Effect on Cardiac Fibroblasts and Protection of Astragaloside IV
GU Jing1,2*, DUAN Yifan1,3, SHU Yafei1,4, HAN Xiaofei1,3
To investigate the effects of exosome-mediated radiation bystander on CFs (cardiac fibroblasts) and the intervention of AST (astragaloside IV), X-exo (radioactive exosomes) were extracted by overspeed centrifugation from rat CFs after 2 Gy X-ray irradiation. Then, the identification of exosome morphology, concentration and surface marker proteins was performed. CFs were divided into control group (Control), irradiation group (X-CFs), co-culture group of CFs and radioactive exosomes (X-exo+CFs), and AST intervention group (X-exo+AST-CFs). Flow cytometry was used to detect cell cycle; Scrape assay was used to detect CFs migration ability; Western blot was used to detect the expression of fibrosis-related molecules TGF-β1 and Col-I. The results showed that, compared with the Control group, the proportion of CFs in G0/G1 phase decreased in X-CFs and X-exo+CFs groups (P<0.01) at 24 h and 48 h after intervention, the proportion of cells in S and G2/M phases increased (P<0.01); the cell mobility of X-CFs and X-exo+CFs groups increased at 12 h, 24 h, 48 h, respectively (P<0.05); the expression of TGF-β1 and Col-I protein in X-CFs groups and X-exo+CFs groups increased at 48 h (P<0.01). Compared with X-exo+CFs group, the proportion of cells in G0/G1 phase increased at 24 h and 48 h in X-exo+AST-CFs group (P<0.01), while S stage and G2/M stage ratio decreased (P<0.01); the cell mobility of X-exo+AST-CFs group decreased by 62.6%, 40.6%, and 41.2% at 12 h, 24 h, and 48 h, respectively (P<0.01); TGF-β1 and Col-I protein expression at 48 h decreased by 15% and 21.9%, respectively (P<0.01). These aforementioned findings indicate that X-ray induced exosomes (radioactive exosomes) can promote the proliferation of CFs, enhance the migration ability of CFs and promote the high expression of fibrosis factors, and AST has some inhibitory effect on the profibrotic effect caused by this exosome-mediated radiation bystander effect.
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