高级检索

    1, 25(OH)2D3通过上调IL-10受体抑制软骨细胞凋亡缓解骨关节炎的分子机制研究

    Molecular mechanism study of 1, 25(OH)2D3 alleviating osteoarthritis by up-regulating IL-10 receptor and inhibiting chondrocyte apoptosis

      摘要
    • 摘要: 目的：探讨骨化三醇〔1, 25（OH）2D3〕对创伤性骨关节炎（PTOA）大鼠膝关节软骨的保护作用及其分子机制。方法：构建PTOA 大鼠模型。将20 只SD 大鼠随机分为对照组、假手术组、PTOA 组和1, 25（OH）2D3组，每组5 只。采用免疫组织化学（IHC）染色法检测膝关节软骨IL-10 受体（IL-10R）和基质金属蛋白酶-13（MMP-13）的表达，番红O 染色法测定大鼠胫骨关节头软骨的损伤程度。分离大鼠骨髓干细胞（BMSCs），将细胞分为未分化组、分化组、1, 25（OH）2D3干预组和1, 25（OH）2D3+IL-10R 抗体组。采用荧光定量PCR（RT-qPCR）法检测细胞中MMP-13和胶原蛋白10a1（Col10a1）mRNA 表达，western blotting法检测磷酸化（p）-JAK、JAK、转化生长因子-β1（TGF-β1）、SOX9、Smad2 和Runx2 蛋白表达，TUNEL 染色法检测细胞凋亡率。结果：IHC 染色显示，与对照组相比，PTOA 组IL-10R 表达水平降低，MMP-13 表达水平升高（均P< 0.05）；与PTOA 组相比，1, 25（OH）2D3组IL-10R 表达水平升高，MMP-13 表达水平降低（均P< 0.05）。番红O 染色结果显示，PTOA 组软骨层厚度低于对照组（P< 0.05），1, 25（OH）2D3组软骨层厚度高于PTOA 组（均P< 0.05）。与分化组相比，1, 25（OH）2D3干预组细胞MMP-13 mRNA相对表达水平降低，Col10a1 mRNA相对表达水平及p-JAK、TGF-β1、SOX9、Smad2和Runx2蛋白表达水平升高（均P< 0.05），TUNEL 阳性细胞率降低（P< 0.05）。与1, 25（OH）2D3干预组比较，1, 25（OH）2D3+IL-10R 抗体组细胞部分逆转了上述指标（P< 0.05）。结论：1, 25（OH）2D3可能通过上调膝关节软骨IL-10R表达，抑制软骨细胞凋亡，从而发挥保护PTOA大鼠膝关节软骨的作用。

       

      Abstract: Objective: To investigate the protective effect and molecular mechanism of calcitriol 1, 25(OH)2D3on knee articular cartilage in rats with post-traumatic osteoarthritis (PTOA).Methods: The PTOA rat model was established.Twenty SD rats were randomly divided into control group, sham operation group, PTOA group and 1, 25(OH)2D3 group, with 5 rats in each group.Immunohistochemistry (IHC) was used to detect the expression of IL-10 receptor(IL-10R)and matrix metalloproteinase-13 (MMP-13)in knee articular cartilage.Safranin O stain-ing method was used to determine the degree of knee articular cartilage injury.Rat bone marrow stem cells(BM-SCs) were isolated and divided into undifferentiation group, differentiation group, 1, 25(OH)2D3 intervention group and 1, 25(OH)2D3+IL-10R antibody group.The expression of MMP-13 and collagen 10a1 (Col10a1)mRNA in cells were detected by real-time fluorescence quantitative PCR (RT-qPCR).Western blotting was used to detect the expression of phospho (p)-JAK, JAK, transforming growth factor-β1 (TGF-β1), SOX9, Smad2 and Runx2 and the apoptosis rate was detected by TUNEL assay.Results: IHC staining results showed that compared with the control group, the expression level of IL-10R decreased and the expression level of MMP-13 increased in the PTOA group (both P< 0.05).Compared with the PTOA group, the expression level of IL-10R increased and the expression level MMP-13 decreased in the 1, 25(OH)2D3 group (both P< 0.05).Safranin O staining results showed that the thickness of cartilage layer in the PTOA group was lower than that in the control group (P< 0.05), and the thickness of cartilage layer in the 1, 25(OH)2D3 group was higher than that in the PTOA group (both P< 0.05).Compared with the differentiation group, the relative expression level of MMP-13 mRNA in the 1, 25(OH)2D3 intervention group decreased, and the relative expression level of Col10a1 mRNA and the pro-tein expression levels of p-JAK, TGF-β1, SOX9, Smad2 and Runx2 increased (all P< 0.05).The rate of TUNEL positive cells decreased(P< 0.05).Compared with the 1, 25(OH)2D3 intervention group, 1, 25(OH)2D3+IL-10R an-tibody group cells partially reversed the above indicators (P< 0.05).Conclusion: The 1, 25(OH)2D3 may play a protective role in knee articular cartilage of rats with PTOA by up-regulating the expression of IL-10R in knee ar-ticular cartilage and inhibiting chondrocyte apoptosis.

       

      • HTML全文
      • 参考文献(16)
      • 相关文章
      • 施引文献
    • 资源附件(0)

    /

    返回文章
    返回