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通过Bingle环加成反应,利用富勒烯与丙二酸二乙酯催化合成羧酸富勒烯C3,经1H-NMR,13C-NMR以及MS-ESI确认其结构和分子量。通过芬顿体系检测了C3清除自由基的能力。利用CCK-8法和Annexin-V/PI染色和流式细胞分析法,分别检测C3对γ射线照射后AHH-1、HIEC细胞存活率,细胞凋亡和细胞周期变化的影响。结果表明,C3具有良好的清除自由基的能力,当芬顿体系中C3终浓度为1000mg/L时,清除效率高达93%左右。对1-12Gyγ射线照射的细胞,C3具有良好的辐射保护作用,表现为照后细胞存活率明显升高,细胞凋亡率显著降低;且对于较高剂量的照射,辐射防护效果较好。说明C3对γ射线照射的细胞具有较好的辐射防护作用,其机理可能主要与其清除自由基的作用有关。
Carboxylic acid fullerene C3 was synthesized by Bingle cycloaddition reaction using fullerene and diethyl malonate. The structure and molecular weight were confirmed by1H-NMR, 13C-NMR and MS-ESI. The ability of C3 to scavenge free radicals was tested by the Fenton system. The effects of C3 on the cell viability, apoptosis and cell cycle changes of AHH-1 and HIEC after γ-ray irradiation were detected by CCK-8 assay and Annexin-V / PI staining and flow cytometry. The results showed that C3 had a good ability to scavenge free radicals. When the concentration of C3 in the Fenton system was 1000 mg / L, the removal efficiency was as high as 93%. For cells irradiated with 1-12Gyγ ray, C3 had a good radiation protection effect, which showed that cell survival rate was significantly increased and cell apoptosis rate was significantly reduced after irradiation. Radiation protection was better for higher doses of irradiation. These results indicate that C3 has a good radiation protection effect on γ-ray irradiated cells, and its mechanism may be mainly related to its function of scavenging free radicals.