阿曼托双黄酮对辐射损伤小鼠骨髓细胞的保护作用

摘要

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.%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12 Gy 60Coγ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。