CDMP-1诱导骨髓间充质干细胞分化成髓核细胞的实验研究  

The experimental study on cartilage derived morphogenetic protein-1 induced differentiation of bone mesenchymal stem cells to nucleus pulposus cells

作　　者：徐斌武 邵斌 朱世杰 叶志炜 彭龙海 李晨[1] 艾江波 陈伟才[1] 程细高[1] 殷明[1] 周荣平[1] 罗军[1] 顾玉荣[1] XU Bin-wu;SHAO Bin;ZHU Shi-jie;YE Zhi-wei;PENG Long-hai;LIChen;AI Jiang-bo;CHEN Wei-cai;CHENG Xi-gao;YIN Ming;ZHOU Rong-ping;LUO Jun;GU Yu-rong(Department of Orthopaedic,the Second Affiliated Hospital of Nanchang University,Nanchang,Jiangxi 330006,China)

机构地区：[1]南昌大学第二附属医院骨科,江西南昌330006 [2]景德镇市第一人民医院骨一科

出　　处：《中国骨与关节损伤杂志》2018年第11期1153-1156,共4页Chinese Journal of Bone and Joint Injury

基　　金：国家自然科学基金(81160226;81460342);江西省自然科学基金(20132BAB205105);江西省教育厅科技支撑计划基金(GJJ12075)

摘　　要：目的 探讨软骨衍生形态发生蛋白-1（CDMP-1）能否诱导骨髓间充质干细胞（BMSCs）向髓核细胞分化,并检测其诱导的细胞是否具有髓核细胞分泌细胞外基质的功能。方法 BMSCs采用0 ng/ml（对照组）以及10 ng/ml（观察组）的CDMP-1处理,将正常椎间盘髓核细胞作为阳性对照组。BMSCs诱导后采用PCR检测Ⅰ型、Ⅱ型胶原基因水平表达,用Western Blot法检测Ⅱ型胶原蛋白水平的表达。采用阿尔辛蓝染色及定量比较蛋白多糖含量的变化。结果 PCR检测结果发现,观察组和对照组中Ⅰ型胶原和Ⅱ型胶原均有增加,而阳性对照组中Ⅱ型胶原明显比观察组更少。基因水平检测结果发现,CDMP-1诱导BMSCs分化后Ⅱ型胶原基因表达比正常髓核细胞还高。观察组中Ⅱ型胶原的蛋白水平表达明显比阳性对照组和对照组细胞更高,对照组中Ⅱ型胶原蛋白水平表达几乎很少。阿尔辛蓝染色结果发现,观察组蛋白多糖含量稍高于阳性对照组,但差异无统计学意义（P〉0.05）;阳性对照组与观察组蛋白多糖含量均高于对照组,差异有统计学意义（P 〈0.05）。结论 CDMP-1能够诱导BMSCs向髓核细胞分化,具有髓核细胞分泌细胞外基质的功能。Objective To investigate the effect of cartilage derived morphogenetic protein-1(CDMP-1) on inducing differentiation of bone mesenchymal stem cells to nucleus pulposus cells. Methods BMSCs were treated with 0 ng/ml(control group) or 10 ng/ml(observed group) CDMP-1, and the nucleus pulposus served as positive group. PCR was used to detect the expression of collagenⅠ and collagenⅡ gene level, and western blot was for detection of the expression of collagenⅡ protein. Alcian Blue staining was used to identify the change of proteoglycan and calculate the difference for the three groups. Results The gene expression of collagen Ⅱ in the observed group was higher than that in the control group and positive group. The collagen Ⅱ protein expression in the observed group was also obviously higher than that in the control group and positive group. The expression of proteoglycan in the observed group for Alcian Blue staining was equal to the positive group and obviously blue than that of the control group, but there was no significant difference between the observe group and control group(P >0.05). There was significant difference in the proteoglycan between the observed group and positive group with the control group(P <0.05). Conclusion CDMP-1 can induce the BMSCs differentiating to nucleus pulposus cells and has the function of secreting extracellular matrix.

关 键 词：软骨衍生形态发生蛋白-1 骨髓间充质干细胞 髓核细胞 Ⅱ型胶原 蛋白多糖 

分 类 号：R-33[医药卫生]