天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (1): 1-4.doi: 10.11958/20160702

• 细胞与分子生物学 •    下一篇

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

薛晓蕾， 韩晓丹， 张俊伶， 田红旗， 樊赛军   

  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 （邮编 300192）
  • 收稿日期:2016-07-19 修回日期:2016-10-30 出版日期:2017-01-15 发布日期:2017-01-15
  • 通讯作者: 樊赛军 E-mail:matchym@126.com
  • 基金资助:
    国家自然科学基金青年项目;中国医学科学院放射医学研究所发展基金项目;中国医学科学院研究生创新基金项目;中国医学科学院中央级公益性科研基金项目

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

XUE Xiao-lei, HAN Xiao-dan, ZHANG Jun-ling, TIAN Hong-qi, FAN Sai-jun   

  1. Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine, Institute of Radiation Medicine, Peking Union
    Medical College and Chinese Academy of Medical Science, Tianjin 300192, China

  • Received:2016-07-19 Revised:2016-10-30 Published:2017-01-15 Online:2017-01-15
  • Contact: FAN Sai-jun E-mail:matchym@126.com
  • Supported by:
    The Youth Program of National Natural Science Fund

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摘要： 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 137Cs γ 射线照射。对照组加 700 μL 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 700 μL 细胞悬液及含 2×10-5 mol/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。

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

Abstract: Abstract: Objective To observe the protective effect of anthocyanin on irradiation induced bone marrow c- kit positive cell injury, and further explore its possible mechanism. Methods Mouse bone marrow c-kit positive cells were collected by cell sorting method. There were 2 groups: control group and anthocyanin group, which were sub-divided into three groups and received 0 Gy, 1 Gy and 4 Gy irradiation respectively. The control group was added 700 μL cell suspension and an equal volume of serum-free hematopoietic stem/progenitor cell culture medium. The 2×10- 5 mol/L anthocyanin was co-cultured with mouse bone marrow c-kit positive cells of anthocyanin group half an hour before irradiation exposure, then cells were cultured for 18 hours under the conventional culture conditions (37 ℃， 5%CO2). Mouse c-kit positive cell viability was measured by bioluminescence, and which was reflected by relative light units (RLU). The ability of colony-forming units was reflected by CFU-GM. The reactive oxygen species (ROS) level and mean fluorescence intensity (MFI) of γ-H2AX were detected by flow cytometry. Results Compared to un-irradiated control group, the cell viability and the number of CFUGM were decreased significantly, while the ROS level and MFI of γ-H2AX were increased in c-kit positive cells irradiated with 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 of

Key words: bone marrow, c-kit positive cell, anthocyanin, ros, DNA damage