花青素对电离辐射引起骨髓c-kit阳性细胞损伤防护作用的体外研究  

The protective effect of anthocyanin on irradiation induced bone marrow c-kit positive cell injury in vitro

作　　者：薛晓蕾[1] 韩晓丹[1] 张俊伶[1] 田红旗[1] 樊赛军[1] 

机构地区：[1]中国医学科学院北京协和医学院放射医学研究所,天津市放射医学与分子核医学重点实验室,300192

出　　处：《天津医药》2017年第1期1-4,共4页Tianjin Medical Journal

基　　金：国家自然科学基金(81402541);国家自然科学基金青年项目(81402633);科技部科研院所技术发展研究专项(2014EG150134);天津科技支撑计划项目(14ZCZDSY00001);中国医学科学院放射医学研究所发展基金项目(1524,1544);中国医学科学院研究生创新基金(1581);中国医学科学院中央级公益性科研基金(1614)

摘　　要：目的观察花青素对电离辐射致小鼠骨髓c-kit阳性细胞损伤的防护作用及可能的作用机制。方法经磁珠细胞分选法获得小鼠骨髓c-kit阳性细胞,分为对照组和花青素组,每组再分成3份,分别接受0、1及4 Gy的137Csγ射线照射。对照组加700μL细胞悬液及等体积的无血清造血干/祖细胞扩增培养基,花青素组加700μL细胞悬液及含2×10-5mol/L花青素的等体积无血清造血干/祖细胞扩增培养基(37℃,5%CO2),照射前30 min加入,照射后继续培养18 h。采用化学发光法检测小鼠骨髓c-kit阳性细胞的细胞活力,以相对荧光强度(RLU)表示。甲基纤维素半固体培养法检测克隆形成数目(CFU-GM)。流式细胞术检测细胞内活性氧(ROS)及磷酸化组蛋白H2AX(γ-H2AX)平均荧光强度。结果组内比较:与0 Gy比较,经1 Gy和4 Gy照射后小鼠骨髓c-kit阳性细胞活力下降,CFU-GM下降,细胞内ROS增加,γ-H2AX平均荧光强度增加(P<0.05)。组间比较:与1 Gy和4 Gy对照组比较,花青素1 Gy和4 Gy组骨髓c-kit阳性细胞活力增加,CFU-GM增加,细胞内ROS降低,DNA损伤程度(γ-H2AX平均荧光强度)降低(P<0.05)。结论花青素通过降低细胞内ROS水平及DNA损伤程度起到对受照射小鼠骨髓c-kit阳性细胞的防护作用。Objective To observe the protective effect of anthocyanin on irradiation induced bone marrow c- kitpositive cell injury, and further explore its possible mechanism. Methods Mouse bone marrow c-kit positive cells werecollected by cell sorting method. There were 2 groups： control group and anthocyanin group, which were sub-divided intothree groups and received 0 Gy, 1 Gy and 4 Gy irradiation respectively. The control group was added 700 μL cell suspensionand an equal volume of serum-free hematopoietic stem/progenitor cell culture medium. The 2×10-5mol/L anthocyanin wasco-cultured with mouse bone marrow c-kit positive cells of anthocyanin group half an hour before irradiation exposure, thencells were cultured for 18 hours under the conventional culture conditions（37 ℃,5%CO2）. Mouse c-kit positive cell viabilitywas measured by bioluminescence, and which was reflected by relative light units（RLU）. The ability of colony-forming unitswas reflected by CFU-GM. The reactive oxygen species（ROS） level and mean fluorescence intensity（MFI） of γ-H2 AX weredetected by flow cytometry. Results Compared to un-irradiated control group, the cell viability and the number of CFU-GM were decreased significantly, while the ROS level and MFI of γ-H2 AX were increased in c-kit positive cells irradiatedwith 1 Gy and 4 Gy（P 〈 0.05）. Compared to 1 Gy and 4 Gy irradiation groups, c-kit positive cell viability and the number ofCFU- GM were increased, the ROS level and MFI of γ- H2 AX were decreased in anthocyanin group（P 〈 0.05）.Conclusion Anthocyanin exhibits a promising protective effect on radiation- induced bone marrow c- kit positive cellinjury, which may be related to the alleviating ROS and DNA damage in bone marrow cells.

关 键 词：骨髓 原花青素类 活性氧 DNA损伤 辐射 电离 小鼠 近交C57BL c-kit阳性细胞 

分 类 号：R811.5[医药卫生—放射医学]