摘要
目的:研究黄芪多糖(APS)对重离子辐照人骨髓间充质干细胞(BMSCs)促增殖作用、维持其基因组不稳定性以及与NF-κB信号通路相关机制。方法:体外培养人BMSCs,随机分为Ctrl组、APS组、IR组、APS+IR组。Ctrl组为常规BMSCs,APS组用50μg/mL APS干预BMSCs,IR组用2Gy 12C6+辐照BMSCs,APS+IR组用50μg/mL APS干预受辐照BMSCs。CCK-8法和克隆形成实验测各组细胞增殖能力;细胞松弛素法测各组BMSCs的微核率;免疫荧光测53BP1免疫荧光簇集焦点;WesternBlot检测P65、p-P65、COX-2蛋白表达。结果:与Ctrl组比较,12C6+辐射后BMSCs增殖水平降低,克隆形成数减少(P<0.05);微核率和免疫荧光焦点显著增多(P<0.01);P65、p-P65、COX-2等相关蛋白上调(P<0.05,P<0.01)。与IR组比较,APS+IR组BMSCs细胞增殖水平升高,克隆形成数增多(P<0.05);微核率和免疫荧光焦点显著减少(P<0.01,P<0.05);P65、p-P65、COX-2蛋白下调(P<0.05)。结论:APS对2Gy 12C6+辐射BMSCs具有促增长作用,可能和下调NF-κB信号通路相关蛋白,维持BMSCs基因组稳定性有关。
Objective:To study the effect of Astragalus Polysaccharide (APS)on the proliferation level and the protective effect on DNA damage of human bone marrow mesenchymal stem cells (BMSCs)and its mechanism related with NF-κB which was induced by heavy Ionizing Radiation.Methods:BMSCs were randomly divided into Control group (Ctrl),Drug group (APS), Radiation group (IR)and Radiation and Drug group (APS+IR).In Ctrl group,BMSCs cultured with normal medium.In APS group,BMSCs was cultured with 50μg/mL APS.In IR group,BMSCs was radiated by 2Gy 12^C^6+ .In APS+IR group,BMSCs was radiated by 2Gy 12^C^6+ and cultured with 50μ g/mL APS.CCK-8 assay and the cell colony formation assay were used to test the proliferation ability of BMSCs.The micronucleus rate was detected by Cytokinesis-block micronucleus assay,the clustered 53BP1 foci was detected by immunofluorescence test.The expression of P65p-P65 and COX-2 was detected by Western Blot.Results: Compared with Ctrl group,the proliferation rate and the colony formation rate of BMSCs decreased in APS+IR group (P<0.05). The cell micronucleus rate and 53BP1 foci level increased significantly (P<0.01).The expression of P65p-P65 and COX-2up-regulated (P<0.05,P<0.01).Compared with IR group,APS promoted the cell proliferation (P<0.05),reduced cell micronucleus rate and the 53BP1 level (P<0.05),and down regulated the expression of P65p-P65 and COX-2(P<0.05).Conclusion:APS can promote the growth of BMSCs irradiated with 2GY 12^C^6+,which may be related to downregulation of NF-κB signaling pathway related proteins and maintenance of genomic stability of BMSCs.
作者
张利英
王磊
张丽昕
张苡铭
许小敏
丁楠
华君瑞
刘永琦
ZHANG Li-ying;WANG Lei;ZHANG Li-xin;ZHANG Yi-ming;XU Xiao-min;DING Nan;HUA Jun-rui;LIU Yong-qi(Provincial Level Key Laboratory for Molecular Medicine of Major Diseases and the Prevention and Treatment with Traditional Chinese Medicine Research in Gansu Colleges and Universities,Gansu University of Traditional Chinese Medicine,Lanzhou 730000,China;Gansu Key Laboratory of Space Radiobiology,Institute of Modem Physics,Chinese Academy of Sciences,Lanzhou 730000,China;Key Laboratory of Dunhuang Medicine and Transformation at Provincial and Ministerial Level,Lanzhou 730000,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2018年第12期5576-5580,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81473457),甘肃中医药大学中青年项目(No.ZQ2014-11).
关键词
重离子
骨髓间充质干细胞
DNA损伤
NF-ΚB
黄芪多糖
Heavy Ionizing
Bone marrow mesenchymal stem cells
DNA damage
NF-κB
Astragalus Polysaccharide
