组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2019, Vol. 15 ›› Issue (6): 389-392.doi: 10.3969/j.issn.1673-0364.2019.06.004

• 论著 • 上一篇    下一篇

三磷酸腺苷对骨髓间充质干细胞向软骨细胞分化的影响及机制研究

雷杰，何承建，贺梦吟   

  1. 湖北省中医院骨伤诊疗中心关节与运动医学科；湖北省中医院中西医结合治疗骨与关节疾病科；湖北省中医院放射科
  • 收稿日期:2019-09-07 修回日期:2019-10-20 出版日期:2019-12-20 发布日期:2019-12-20
  • 作者简介:雷杰(E-mail:xue582581581481147@163.com)。

Effect and Mechanism of Adenosine Triphosphate on Differentiation of Bone Marrow Mesenchymal Stem Cells into Chondrocytes

LEI Jie,HE Chengjian,HE Mengyin   

  1. Department of Joint and Sports Medicine,Bone Trauma Diagnosis and Treatment Center, Hubei Traditional Chinese Medicine Hospital;Treatment of Bone and Joint Diseases by Integrated Traditional Chinese and Western Medicine, Hubei Traditional Chinese Medicine Hospital;Radiology Department, Hubei Traditional Chinese Medicine Hospital
  • Received:2019-09-07 Revised:2019-10-20 Online:2019-12-20 Published:2019-12-20

摘要： 目的探讨三磷酸腺苷(Adenosine triphosphate,ATP)对骨髓间充质干细胞(BMSCs)向软骨细胞分化的影响及其作用机制。方法取4～5周龄SD大鼠的BMSCs,体外培养至第3代。以PNPP法检测不同浓度ATP(150μmol/L、300μmol/L、600μmol/L)及BBG(P2X7受体抑制剂)对BMSCs碱性磷酸酶表达的影响。采用RT-PCR和real time PCR法检测ATP及BBG对BMSCs多个成软骨诱导指标mRNA表达的影响。结果加入ATP后,BMSCs碱性磷酸酶表达明显增加(P<0.05),其中ATP浓度为150μmol/L和300μmol/L时碱性磷酸酶表达量最高(P<0.05);多个成软骨指标,如SOX6、SOX9及COL2A1等的表达水平提高(P<0.05);加入P2X7受体抑制剂后,ATP对碱性磷酸酶的促进作用降低(P<0.05),成软骨指标SOX6、SOX9及COL2A1等的表达亦明显减少(P<0.05)。结论一定浓度的ATP对BMSCs向软骨细胞分化具有促进作用,可能是通过介导P2X7受体而发挥作用。

关键词: 三磷酸腺苷, 骨髓间充质干细胞, 成软骨分化

Abstract: Objective To investigate the effect of adenosine triphosphate on the differentiation of BMSCs into chondrocytes and its mechanism. Methods The primary cells of BMSCs from 4-5 weeks old SD rats were cultured to the third generation in vitro. PNPP method was used to detect the effects of ATP with different concentrations(150, 300 and 600μmol/L) and BBG(receptor inhibitor of P2X7) of on alkaline phosphatase expression in BMSCs. The mRNA expression of multiple cartilage markers was detected by semi-quantitative reverse transcription polymerase chain reaction and real time PCR. Results After ATP was added, the expression of alkaline phosphatase in BMSCs was significantly increased(P<0.05).The highest expression of alkaline phosphatase was observed when ATP concentration was 150 and 300 μmol/L(P<0.05).The expression levels of indicators such as SOX6, SOX9 and COL2A1 were increased(P<0.05). After the addition of P2X7 receptor inhibitor, the promotion effect of ATP on alkaline phosphatase decreased(P <0.05), and the expressions of chondrogenic indicators such as SOX6, SOX9 and COL2A1 were also significantly decreased(P <0.05). Conclusion A certain concentration of ATP can promote the differentiation of bone marrow mesenchymal cells into chondrocytes, which may play a role through the mediation of P2X7 receptor.

Key words: Adenosine triphosphate, Bone marrow derived mesenchymal stem cells, Chondrogenic differentiation

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