吲哚-3-甲醇对小鼠骨髓造血细胞辐射损伤的防护作用及其机制  

The effect and mechanism of indole-3-carbinol on alleviating the radiation-induced bone marrow hematopoietic cell injury in mice

作　　者：路璐[1] 董佳丽[1] 樊赛军[1] LU Lu;DONG Jia-li;FAN Sai-jun(Institute of Radiation Medicine, Chinese Academy of Medical Science and Peking Union Medical Collage, Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine, Tianjin 300192, China)

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2期155-159,共5页Tianjin Medical Journal

基　　金：国家自然科学基金面上项目(81071906;81172127;81572969);天津市应用基础与前沿技术研究计划青年项目(15JCQNJC46000);"协和青年基金资助";"中央高校基本科研业务费专项资金资助"(3332013044);中国医学科学院;北京协和医学院"中央级公益性科研院所基本科研业务费"(2016RC310016)

摘　　要：目的研究吲哚-3-甲醇(I3C)对小鼠骨髓造血细胞辐射损伤的保护作用及机制。方法 (1)密度梯度离心法获得CD45.1亚型C57BL/6J小鼠骨髓有核细胞,经0 mol/L、10^(-8)mol/L^10^(-3)mol/L I3C处理后,接受不同剂量(0Gy、1 Gy、4 Gy)的^(137)Csγ-射线照射;继续培养18 h后采用生物发光法检测细胞活力。(2)设空白对照组和10^(-6)mol/LI3C组,经上述3种剂量射线照射后,接种于甲基纤维素半固体培养基中培养7 d,观察骨髓粒-单核巨噬细胞集落(CFU-GM)形成情况。(3)取24只CD45.2亚型小鼠接受8 Gy137Csγ-射线照射作为受体,CD45.1亚型小鼠骨髓有核细胞(供体)设空白对照组、4 Gy照射组和4 Gy照射+10^(-6)mol/L I3C组。将供体与竞争者(CD45.2亚型)骨髓细胞混和后,接种于受体小鼠体内(每组8只),流式细胞术检测受体小鼠外周血细胞中供体来源的细胞比例。(4)细胞设空白对照组、10^(-6)mol/L I3C组、1 Gy照射组和1 Gy照射+10^(-6)mol/L I3C组。培养24 h后收集细胞,提取蛋白后Western blot法检测各组核因子NF-E2相关因子(Nrf2)、血红素加氧酶(HO)-1表达。结果 (1)I3C浓度>10-4mol/L时出现明显的细胞毒性作用(P>0.05);相同剂量射线照射下,10^(-7)~10^(-6)mol/L I3C可减轻射线对细胞的损伤;因此选取10^(-6)mol/L为本研究I3C的实验浓度。(2)相同剂量射线照射下,10^(-6)mol/L I3C组CFU-GM形成数量较空白对照组明显升高(P<0.05)。(3)流式细胞结果显示,4 Gy照射组细胞移植后,受体小鼠外周血中供体细胞比例较对照组明显降低(P<0.05),而10^(-6)mol/L I3C预处理的供体小鼠细胞移植后,受体小鼠中供体细胞比例较4 Gy照射组升高(P<0.05)。(4)Western blot结果显示,1 Gy照射+10^(-6)mol/L I3C组Nrf2和HO-1蛋白表达水平明显高于其他3组(P<0.05)。结论 I3C可以减轻辐射引起的小鼠造血细胞损伤和功能下降,其机制可能与激活Nrf2/HO-1通路有关。Objective To investigate the protective effect of indole-3-carbinol(I3C)on radiation-induced mousebone marrow hematopoietic cell injury and the involved mechanisms.Methods(1)The bone marrow nuclear cells(BMNCs)from CD45.1subtype of C57BL/6J mice were collected by a density gradient centrifugation method.The BMNCswere pretreated with a series doses of I3C(0mol/L,10-8mol/L-10-3mol/L)and then exposed with radiation of137Csγ-ray(doses of irradiation were0Gy,1Gy and4Gy).After18-hour culturing,the bioluminescence method was used to detect thecell viability.(2)These cells were divided into control group and10-6mol/L I3C group.Both groups were received theirradiation(0Gy,1Gy and4Gy)and inoculated into the methylcellulose semi-solid culture medium to incubate7days,thecolony forming unit-granulocyte monocytes(CFU-GM)were observed.(3)Twenty-four CD45.2subtype mice used as the receptor were exposed with8Gy radiation.The CD45.1BMNCs were divided into control group,4Gy irradiation group,4Gyirradiation and10-6mol/L I3C group.Donor cells were harvested from C57BL/6J(CD45.1)mice after they received varioustreatments,and were then mixed with competitive BMNCs from C57BL/6J(CD45.2)mice.The mixed cells were transplantedinto recipient mice(8mice/group).Flow cytometry was used to analyze the proportion of donor cells in peripheral blood ofreceptor.(4)The cells were divided into control group,10-6mol/L I3C group,1Gy irradiation group,1Gy irradiation with10-6mol/L I3C group.After24-hour culturing,Western blot assay was used to detect the expression levels of nuclear factorerythroid2-related factor2(Nrf2)and hemeoxygenase-1(HO-1).Results(1)I3C showed a significant cytotoxic effect onthe BMNCs when its concentration was above10-4mol/L.10-7-10-6mol/L I3C could reduce the radiation injury of BMNCsunder the same dose of irradiation.Therefore,10-6mol/L I3C was chosen for subsequent experiments.(2)The CFU-GM wassignificantly higher in10-6mol/L I3C group than that of control group(P<0.05).(3)Results of flow cytometry showed thatthe proportion

关 键 词：骨髓祖代细胞 辐射损伤 辐射防护 造血干细胞 造血祖细胞 吲哚-3-甲醇 

分 类 号：R331.2[医药卫生—人体生理学]