骨形态发生蛋白7抑制椎间盘髓核细胞的凋亡  被引量:3

Bone morphogenetic protein 7 inhibits apoptosis of nucleus pulposus cells

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作  者:徐学振 宋立先 李爱群 杨坚 李晓鲲 王占青 石青鹏 Xu Xuezhen;Song Lixian;Li Aiqun;Yang Jian;Li Xiaokun;Wang Zhanqing;Shi Qingpeng(Yantai Affiliated Hospital of Binzhou Medical University,Yantai 264100,Shandong Province,China)

机构地区:[1]滨州医学院烟台附属医院,山东省烟台市264100

出  处:《中国组织工程研究》2022年第17期2726-2731,共6页Chinese Journal of Tissue Engineering Research

基  金:山东省医药卫生科技发展计划项目(2019WS340),项目名称:不同代次兔髓核种子细胞超低温保存的实验研究,项目负责人:徐学振。

摘  要:背景:椎间盘退变是引起下腰痛的主要原因之一,目前临床的手术治疗可以缓解疼痛症状,但会牺牲椎间盘的功能,无法完全治愈椎间盘退变,开发新的生物学治疗方法对于遏制早期椎间盘退变的进展,或再生退变的椎间盘组织是非常重要的。目的:探讨骨形态发生蛋白7能否减轻人椎间盘髓核细胞在传代培养过程的细胞衰老及其可能机制。方法:以行椎间盘摘除术的患者为研究对象,手术过程中获取椎间盘组织并提取人椎间盘髓核细胞体外传代培养6次。在培养基中外源性加入骨形态发生蛋白7或磷酯酰肌醇-3激酶特异性抑制剂LY294002,分别检测细胞增殖能力、端粒酶活性、β-半乳糖苷酶活性,以及细胞凋亡相关基因和磷酯酰肌醇-3激酶/蛋白激酶B信号通路分子的表达情况。结果与结论:(1)细胞培养基中添加骨形态发生蛋白7后可以显著提高髓核细胞增殖能力和端粒酶活性,降低细胞β-半乳糖苷酶活性,并下调p16和p53的表达,激活磷酯酰肌醇-3激酶/蛋白激酶B信号通路;(2)此外通过磷酯酰肌醇-3激酶特异性抑制剂LY294002能够明显逆转骨形态发生蛋白7对髓核细胞的抗凋亡作用;(3)提示骨形态发生蛋白7通过激活磷酯酰肌醇-3激酶/蛋白激酶B途径发挥髓核细胞的抗凋亡作用,可为异体椎间盘髓核细胞的体外培养和组织工程学研究提供新思路。BACKGROUND:Disc degeneration is one of the major causes of lower back pain,and current clinical surgical treatments can relieve painful symptoms but sacrifice disc function and cannot completely cure disc degeneration.Therefore,it is important to develop new biological treatments to curb the progression of early disc degeneration,or to regenerate degenerated disc tissue.OBJECTIVE:To investigate whether bone morphogenetic protein 7 can reduce the senescence of human nucleus pulposus cells during subculture and its possible mechanism.METHODS:Patients who underwent discectomy were enrolled in this study.During the operation,the intervertebral disc tissue was obtained and human nucleus pulposus cells were extracted and subcultured in vitro for six times.The cells were then cultured with bone morphogenetic protein 7 or LY294002,a specific inhibitor of phosphatidyl-inositol 3-kinase.Cell proliferation ability,telomerase activity,β-galactosidase activity,and the expression of apoptosis-related genes and phosphatidyl-inositol 3-kinase/protein kinase B signaling pathway molecules were detected.RESULTS AND CONCLUSION:Bone morphogenetic protein 7 could significantly increase the cell proliferation and telomerase activity of nucleus pulposus cells,decrease the activity ofβ-galactosidase,down-regulate the expression of p16 and p53,and activate phosphatidyl-inositol 3-kinase/protein kinase B signaling pathway.In addition,LY294002 could significantly reverse the anti-apoptotic effect of bone morphogenetic protein 7 on nucleus pulposus cells.Overall,bone morphogenetic protein 7 exerts the anti-apoptotic effect on nucleus pulposus cells by activating the phosphatidyl-inositol 3-kinase/protein kinase B signaling pathway.The results of this study can provide new knowledge for in vitro culture and tissue engineering of allogenic nucleus pulposus cells.

关 键 词:椎间盘再生 髓核细胞 骨形态发生蛋白7 细胞凋亡 

分 类 号:R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]

 

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