UTX表观调控骨髓间充质干细胞修复椎间盘纤维环缺损作用机制的体外实验研究
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1.内蒙古医科大学第二附属医院脊柱外科中心C区;2.内蒙古医科大学第二附属医院急诊科;3.内蒙古国际蒙医医院手术麻醉科

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内蒙古医科大学科技百万工程课题 [YKD2018KJBW(LH)006];内蒙古自治区自然科学基金项目(2019BS03034)


UTX epigenetically regulates the effect of BMSCs to repair the disc fibroring defect in vitro experimental studies of the mechanism
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Affiliation:

1.District C, Spinal Surgery Center, the Second Affiliated Hospital of Inner Mongolia Medical University;2.Department of Emergency, the Second Affiliated Hospital of Inner Mongolia Medical University,;3.Department of Surgical Anesthesia, Inner Mongolia International Mongolian Hospital,;4..District C, Spinal Surgery Center, the Second Affiliated Hospital of Inner Mongolia Medical University,

Fund Project:

Engineering Project on Science and Technology of Inner Mongolia Medical University, No. YKD2018KJBW(LH)006 ; Natural Science Foundation of Inner Mongolia Autonomous Region, No. 2019BS03034

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    摘要:

    [目的] 为了研究表观调控骨髓间充质干细胞中UTX基因对于骨髓间充质干细胞的作用机制,为下一步进行椎间盘纤维环缺损的修复奠定基础。[方法]利用密度梯度离心法和细胞悬浮法从小鼠中分离提取原代骨髓间充质干细胞,然后进行CD29、CD34、CD44表面抗原流式细胞学鉴定,进行传代培养,培养至第三代生长活跃的骨髓间充质干细胞。利用慢病毒载体构建UTX上调组、UTX下调组和UTX空载组病毒,进行转染骨髓间充质干细胞,并进行QRT-PCR检测,检测转染效率,将干预后的三组骨髓间充质干细胞进行成骨和成软骨实验,分析观察不同组成骨和成软骨情况。由于椎间盘纤维环主要由II型胶原构成,最后进行II型胶原免疫荧光检测,检测不同分组II型胶原的分布以及荧光定量情况。[结果]UTX上调组与UTX下调组相比,更具有成骨和成软骨能力。在II型胶原细胞免疫荧光中,UTX上调组中II型胶原的分布以及荧光表达情况高于UTX下调组。[结论]过表达骨髓间充质干细胞中的UTX基因,促使成骨与成软骨的发生,促使II型胶原产生更多与UTX下调组相比。可以促使椎间盘纤维环缺损的修复。

    Abstract:

    [Objective] To study the mechanism of UTX gene in MSCs and to lay the foundation for the repair of disc fibroring defect.[Methods] Primary MSCs were isolated from mice using density gradient centrifugation and cell suspension, followed by surface antigen flow cytology identification of CD29, CD34,CD44, and passaged to the third generation of active-growing MSCs.The UTX up-regulated, UTX down-regulated and UTX were transfected with QRT-PCR to detect transfection efficiency, The three groups of bone marrow mesenchymal stem cells after intervention were tested for osteogenesis and chondrogenesis analying and observing different bone composition and cartilage formation. Since the disc fiber ring is mainly composed of collagen II. Finally, immunofluorescence detection of type II collagen was performed to detect the distribution and fluorescence quantification of type II collagen in different groups. [Results] The UTX up-gulated group was more osteogenic and chondrogenic compared to the UTX down-regulated group.In the type II collagen cell immunofluorescence, the distribution of type II collagen and the fluorescence expression was higher in the UTX upregulated group than in the UTX down-regulated group.[Conclusions] Overexpression of UTX gene in bone marrow mesenchymal stem cells promotes osteogenesis and chondrogenesis, and promotes more type II collagen production compared with the down-regulated UTX group. It can promote the repair of intervertebral disc annulus fibrosus defect.

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  • 收稿日期:2021-11-30
  • 最后修改日期:2021-12-01
  • 录用日期:2021-12-16
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