阿曼托双黄酮对辐射损伤小鼠骨髓细胞的保护作用

Protective effect of amentoflavone on radiation-reduced damage in mice bone marrow

目的探讨阿曼托双黄酮(amentoflavone,AF)对辐射小鼠骨髓细胞的防护作用及可能机制。方法培养C57BL/6小鼠骨髓原代细胞,分为正常对照组、照射对照组、AF 2.5和5μmol/L组,每组3复孔,给药12 h后换成普通培养基,给予12Gy60Coγ射线一次性照射,于照后6和12 h分别用Hoechst染色法检测细胞的凋亡,用PI染色和流式细胞仪检测细胞周期,用酶联免疫吸附法检测细胞培养基中TNF-α的浓度。结果 AF对辐射后小鼠骨髓细胞凋亡及细胞周期变化无明显影响,但能显著降低辐射后细胞上清中TNF-α的浓度,且对照后6 h细胞作用明显(2.5μmol/L组P<0.05,5μmol/L组P<0.01);照后12 h也具有同样趋势,与照射对照组相比,5μmol/L组细胞上清中TNF-α表达明显降低(P<0.05)。结论 AF对C57BL/6小鼠骨髓细胞具有一定的辐射防护作用,它可能是通过抑制辐射后骨髓细胞TNF-α的表达来发挥辐射防护作用。

Objective To investigate the protective effect and possible mechanism of amentoflavone （AF） on bone marrow cells of mice injured by irradiation. Methods Primary bone marrow cells of male C57BL/6 mice were cultured and randomly divided into 4 groups （normal control, radiation control,AF 2.5μmol/L and 5μmol/L）, with 3 samples in each group. After treated with AF for 12 h, the cells were injured by 12 Gy 60Coγirradiation. 6 h and 12 h post-irradiation, apoptosis was evaluated by using Hoechst 33258 stain, cell cycle was determined by flow cytometry while the level of TNF-α was tested by ELISA. Results The cell cycle arrest and apoptosis were not significantly affected by Amentoflavone. Amentoflavone （5 μmol/L）could significantly inhibit the production of TNF-α on cell supernatant of mouse bone marrow cells at 6 h or 12 h after radiation and 2.5 μmol/L Amentoflavone could significantly inhibit the production of TNF-α at 6 h after radiation. Conclusion Taken together, the data suggest that AF may have radioprotection against damage in mice bone marrow by inhibiting the production of TNF-α.