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    顾蓉蓉, 丁长伟. miRNA-200c/SOX2轴在实验性牙周炎牙槽骨吸收中的作用研究[J]. 蚌埠医科大学学报, 2023, 48(3): 281-285. DOI: 10.13898/j.cnki.issn.1000-2200.2023.03.001
    引用本文: 顾蓉蓉, 丁长伟. miRNA-200c/SOX2轴在实验性牙周炎牙槽骨吸收中的作用研究[J]. 蚌埠医科大学学报, 2023, 48(3): 281-285. DOI: 10.13898/j.cnki.issn.1000-2200.2023.03.001
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    GU Rong-rong, DING Chang-wei. Study on the role of miRNA-200c/SOX2 axis in alveolar bone resorption in experimental periodontitis[J]. Journal of Bengbu Medical University, 2023, 48(3): 281-285. DOI: 10.13898/j.cnki.issn.1000-2200.2023.03.001
    Citation: GU Rong-rong, DING Chang-wei. Study on the role of miRNA-200c/SOX2 axis in alveolar bone resorption in experimental periodontitis[J]. Journal of Bengbu Medical University, 2023, 48(3): 281-285. DOI: 10.13898/j.cnki.issn.1000-2200.2023.03.001
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    miRNA-200c/SOX2轴在实验性牙周炎牙槽骨吸收中的作用研究

    Study on the role of miRNA-200c/SOX2 axis in alveolar bone resorption in experimental periodontitis

      摘要
    • 摘要:
      目的探讨miRNA-200c/转录因子Y框蛋白2(SOX2)轴在实验性牙周炎牙槽骨吸收中的作用。
      方法将C57BL/6小鼠随机分为空白组、模型组、miRNA-200c组和联合组。模型组、miRNA-200c组和联合组小鼠分别给予慢病毒空载体、miRNA-200c慢病毒过表达载体及miRNA-200c和SOX2慢病毒过表达载体。以牙龈卟啉单胞菌涂抹模型组、miRNA-200c组和联合组小鼠口腔建立慢性牙周炎模型。检测各组牙周组织miRNA-200c和SOX2表达水平、牙龈指数、釉牙骨质界(CEJ)到牙槽嵴顶(ABC)距离。HE染色分析小鼠牙周组织病理形态学变化。qPCR检测小鼠牙周组织中miRNA-200c和SOX2表达水平。Western blotting检测小鼠牙周组织中骨保护素(OPG)、核因子κB受体活化因子配体(RANKL)、核因子κB受体因子(RANK)蛋白表达水平。
      结果与空白组相比，模型组小鼠牙周组织中miRNA-200c表达水平降低(P < 0.05)，SOX2表达水平升高(P < 0.05)；miRNA-200c组牙周组织中miRNA-200c表达水平高于空白组和模型组，SOX2表达水平低于模型组(P < 0.05)；联合组牙周组织中miRNA-200c及SOX2表达水平均高于空白组和模型组(P < 0.05)。模型组、miRNA-200c组及联合组牙龈指数及CEJ至ABC距离高于空白组(P < 0.05)，miRNA-200c组牙龈指数及CEJ至ABC距离均低于模型组(P < 0.05)，联合组牙龈指数及CEJ至ABC距离均高于miRNA-200c组(P < 0.05)。Western blotting结果显示，模型组、miRNA-200c组及联合组RANKL、RANK表达水平及RANK/OPG比值均高于空白组(P < 0.05)，miRNA-200c组RANKL、RANK表达水平及RANK/OPG比值均低于模型组(P < 0.05)，联合组RANKL、RANK表达水平及RANK/OPG比值均高于miRNA-200c组(P < 0.05)。HE染色结果显示，模型组小鼠牙周组织出现明显大量炎症细胞浸润，胶原纤维排列紊乱、变性、断裂，而miRNA-200c组小鼠这些症状相比于模型组得到显著缓解，但是联合组小鼠相比于miRNA-200c组小鼠也出现了显著的炎症细胞浸润，胶原纤维排列紊乱、变性、断裂现象。荧光素酶活性检测显示，miRNA-200c mimic能明显降低SOX2野生型荧光素酶活性(P < 0.01)，而对SOX2突变型荧光素酶活性影响差异无统计学意义(P>0.05)。
      结论miRNA-200c能改善小鼠牙周炎炎症反应及牙槽骨吸收情况，可能与SOX2调控的RANKL/RANK/OPG轴有关。

       

      Abstract:
      ObjectiveTo investigate the role of miRNA-200c/sex determining region Y-box 2 (SOX2) axis in alveolar bone resorption in experimental periodontitis.
      MethodsC57BL/6 mice were randomly divided into blank group, model group, miRNA-200c group and combination group.Mice in the model group, miRNA-200c group and combination group were given lentiviral empty vector, miRNA-200c lentiviral overexpression vector, miRNA-200c plus SOX2 lentiviral overexpression vector, respectively.Chronic mice periodontitis models were established by smearing Porphyromonas gingivalis in the oral cavity of mice in model group, miRNA-200c group and combination group.The expression levels of miRNA-200c and SOX2, gingival index, and distance from the cementoenamel junction (CEJ) to the alveolar bone crest (ABC) were determined in the periodontal tissues of mice in all groups.HE staining was used to analyze the pathomorphological changes of periodontal tissues of mice.The expression levels of miRNA-200c and SOX2 in periodontal tissues of mice were detected by qPCR.The protein expression levels of osteoprotegerin (OPG), receptor activator of nuclear factor-κB ligand (RANKL), and receptor activator of nuclear factor-κB (RANK) in periodontal tissues of mice were detected by Western blotting.
      ResultsCompared with the blank group, the expression level of miRNA-200c in the periodontal tissue of the model group decreased (P < 0.05), and the expression level of SOX2 increased (P < 0.05).The expression level of miRNA-200c in the periodontal tissue of the miRNA-200c group was higher than that of the blank group and model group, and the expression level of SOX2 was lower than that of the model group (P < 0.05).The expression levels of miRNA-200c and SOX2 in the periodontal tissue of the combination group were higher than those of the blank group and model group (P < 0.05).The gingival index and the distance from CEJ to ABC in the model group, miRNA-200c group and combination group were higher than those in the blank group (P < 0.05), which in the miRNA-200c group were lower than those in the model group (P < 0.05), and which in the combination group were higher than those in the miRNA-200c group (P < 0.05).The results of Western blotting showed that the expression levels of RANKL and RANK, and the ratio of RANK/OPG in the model group, miRNA-200c group and combination group were higher than those in the blank group (P < 0.05), which in the miRNA-200c group were lower than those in the model group (P < 0.05), and which in the combination group were higher than those in the miRNA-200c group (P < 0.05).The results of HE staining showed that a large number of inflammatory cells infiltrated in the periodontal tissue of mice in the model group, and collagen fibers were disordered, denatured and broken; compared with the model group, these symptoms in the miRNA-200c group were significantly alleviated; compared with the miRNA-200c group, mice in the combination group also showed significant inflammatory cell infiltration, and collagen fibers were disordered, denatured and broken.The detection of luciferase activity showed that miRNA-200c mimic could significantly reduce the activity of wild-type SOX2 luciferase (P < 0.01), but there was no significant difference in the activity of mutant SOX2 luciferase (P>0.05).
      ConclusionsMiRNA-200c can improve the inflammatory response and alveolar bone resorption in mice with periodontitis, which may be related to the RANKL/RANK/OPG axis regulated by SOX2.

       

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