条件性敲除骨髓间充质干细胞中 3- 磷酸肌醇依赖蛋白激酶 1 基因后的 成骨细胞分化.

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    • Alternate Title:
      Osteoblast differentiation after conditional knockout of 3-phosphoinositide-dependent protein kinase-1 gene from bone marrow mesenchymal stem cells.
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    • Abstract:
      BACKGROUND: The research on 3-phosphoinositide-dependent protein kinase-1 (PDK-1) in and outside China is mainly concentrated in endocrinology and oncology. There are no systematic studies and reports on its influence on osteogenic differentiation in orthopedics. OBJECTIVE: To observe the role of PDK-1 in osteoblast differentiation by transfecting PDK-1flox/flox mouse bone marrow mesenchymal stem cells with adenovirus stably expressing cre enzyme (pHBAd-cre-EGFP).METHODS: Bone marrow mesenchymal stem cells from homozygous PDK-1flox/flox mice were obtained and cultured in vitro. A control group, an empty virus group (pHBAd-EGFP) and a pHBAd-cre-EGFP group were set. In the control group, osteoblast induction medium was used to induce bone marrow mesenchymal stem cells. In pHBAd-EGFP group, pHBAd-EGFP was used to transfect bone marrow mesenchymal stem cells. In the pHBAd-cre-EGFP group, the recombinant adenovirus containing Cre recombinant enzyme (pHBAd-cre-EGFP) was used to transfect with the PDK-1 gene in bone marrow mesenchymal stem cells followed by osteogenic induction. Alkaline phosphatase staining, alkaline phosphatase activity assay, alizarin red staining, and qPCR were used to detect osteogenesis-related gene expression and to evaluate the differentiation and maturation of osteoblasts. RESULTS AND CONCLUSION: The alkaline phosphatase secretion, alkaline phosphatase activity, and the ability of cell mineralization in the pHBAd-cre-EGFP group were significantly lower than those in the other two groups (P < 0.05, P < 0.01), and the expression levels of Runx2, osteocalcin, and collagen I were also significantly lower than those in the other two groups (P < 0.01). The results have confirmed that interference with PDK-1 gene in vitro can significantly inhibit the osteogenic differentiation of bone marrow mesenchymal stem cells. [ABSTRACT FROM AUTHOR]
    • Abstract:
      背景:国内外对于3-磷酸肌醇依赖蛋白激酶1(3-phosphoinositide-dependent protein kinase-1,PDK-1)的研究主要集中在内分泌和肿瘤学等学 科领域,在骨科学中关于其对成骨分化的影响尚未有系统研究与报道。 目的:通过使用稳定表达cre酶的腺病毒(pHBAd-cre-EGFP)转染PDK-1flox/flox小鼠骨髓间充质干细胞,观察PDK-1在成骨细胞分化中的作用。 方法:体外培养获得来源于纯合子PDK-1flox/flox小鼠的骨髓间充质干细胞,设立对照组、空载病毒组(pHBAd-EGFP)和pHBAd-cre-EGFP组,对照 组仅用成骨细胞诱导培养基诱导培养骨髓间充质干细胞,空载病毒组使用pHBAd-EGFP转染骨髓间充质干细胞,pHBAd-cre-EGFP组使用含 Cre重组酶的重组腺病毒(pHBAd-cre-EGFP)干扰骨髓间充质干细胞中的PDK-1基因,然后进行成骨诱导,采用碱性磷酸酶染色、碱性磷酸酶 活性、茜素红染色和qPCR检测成骨相关基因表达来评价各组成骨细胞的分化成熟情况。 结果与结论:pHBAd-cre-EGFP组细胞的碱性磷酸酶分泌、碱性磷酸酶活性、矿化能力均明显低于其他2组(P < 0.05,P < 0.01),成骨细胞相 关基因Runx2、骨钙素和Ⅰ型胶原的表达也明显低于其他2组(P < 0.01)。结果表明,体外干扰PDK-1基因表达可显著抑制骨髓间充质干细胞 成骨分化。 [ABSTRACT FROM AUTHOR]
    • Abstract:
      Copyright of Chinese Journal of Tissue Engineering Research / Zhongguo zu zhi gong cheng yan jiu is the property of Chinese Journal of Tissue Engineering Research and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)