衰老骨髓间充质干细胞中端粒短缩与成骨能力下降相关性研究  

The abbreviation of telomere and the decline of osteogenic differentiation in senescent BMSCs:A correlation study

作　　者：林源[1] 陶天遵[1] 陶树清[1] 王声雨[1] 李超[1] LIN Yuan;TAO Tianzun;TAO Shuqing;WANG Shengyu;LI Chao(Department of Orthopedics,The 2nd Affiliated Hospital of Harbin Medical University,Harbin 150081,China)

机构地区：[1]哈尔滨医科大学附属第二医院,黑龙江哈尔滨150081

出　　处：《中国骨质疏松杂志》2018年第10期1317-1323,共7页Chinese Journal of Osteoporosis

摘　　要：目的骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)是组织工程中修复骨和软骨缺损的重要种子细胞。老年供体BMSCs的增殖速度和分化能力均低于年轻供体,其相关机制仍不清楚,希望通过相关实验对其进行探究。方法采用SD大鼠的BMSCs进行培养,并通过应用H_2O_2处理和连续传代诱导细胞衰老。通过β-半乳糖苷酶染色和端粒长度分析检测衰老,同时对细胞增殖和成骨分化能力进行检测。应用Olaparib维持端粒长度来研究端粒长度在细胞衰老、增殖和分化能力方面的影响。结果 H_2O_2可以增加BMSCs中β-半乳糖苷酶染色阳性率、缩短端粒长度、降低细胞增殖率和碱性磷酸酶活性。BMSCs长期传代后也可观察到成骨分化的衰老。抑制端粒长度下降可降低H_2O_2诱导所致的β-半乳糖苷酶染色阳性率增加,促进细胞增殖,增强成骨分化能力。结论端粒长度下降可引起BMSCs的衰老,从而降低其成骨分化能力、抑制干细胞增殖。Objective Bone marrow mesenchymal stem cells(BMSCs)are important seed cells for the repair of bone and cartilage defect in the tissue engineering.The proliferation rate and differentiation capacity of BMSCs from the old donors may be less than that from young donors.However,the related mechanism remains unclear.Methods BMSCs in Sprague Dawley(SD)rats were cultured.Hydrogen peroxide(H 2O 2)and continual passage of BMSCs were used to induce senescence.Senescence was detected with the SAβ-Gal staining and the telomere length analysis.Cell proliferation and osteogenic differentiation were also observed.Olaparib was used to maintain the telomere length and the role of telomere length and senescence on the cell proliferation and differentiation were investigated.Results H 2O 2 increased the positive rate of SA-β-Gal staining in BMSCs and shortened the length of the telomere.The proliferation rate and ALP activity were also decreased by the H 2O 2.The senescence and decline of osteogenic differentiation could also be observed after prolonged passage of BMSCs.Inhibition of telomere length decline could attenuate the increased positive rate of SA-β-Gal staining induced by H 2O 2,promote the cell proliferation,and enhance the capacity of osteogenic differentiation.Conclusion Senescence can be induced by the decline of telomere length and can reduce the capacity of osteogenic differentiation and inhibit cell proliferation in BMSCs.

关 键 词：骨髓间充质干细胞 衰老 端粒 成骨分化 动物实验 

分 类 号：R-332[医药卫生]