小鼠牙周炎模型中骨膜素的时空表达分析

Yue Li, Chunmei Xu, Xudong Xie, Peilei Shi, Jun Wang, Yi Ding
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引用次数: 0

摘要

研究目的本研究旨在探讨牙周炎症期间小鼠牙周膜组织蛋白表达的时空变化:方法:用丝线结扎法构建牙周炎模型。小鼠随机分为五组,包括对照组、结扎 4 天组、结扎 7 天组、结扎 14 天组和自愈组(结扎 14 天后拔除丝线 14 天)。对各组小鼠牙周组织的动态变化进行显微 CT 和组织学染色。使用 RNAscope 和免疫组化染色法分析牙周炎不同阶段的骨膜素变化规律。细胞实验分为三组:对照组、脂多糖(LPS)刺激组(用 LPS 处理 12 小时)和 LPS 刺激去除组(用 LPS 处理 3 小时后用培养基培养 9 小时)。采用实时定量聚合酶链反应(qRT-PCR)检测骨膜蛋白、转化生长因子-β1(TGF-β1)和基质金属蛋白酶2(MMP2)的表达:结扎 7 天后观察到牙槽骨明显吸收。随着结扎时间的延长,小鼠牙周组织的损伤加剧,表现为破骨细胞增多、牙周膜间隙增宽和牙槽骨高度降低。在自愈组中观察到了一定程度的牙周组织修复。与对照组相比,在 4 天和 7 天时,牙周膜生长因子的表达量减少;与 4 天和 7 天相比,在 14 天时,牙周膜生长因子的表达量增加。自我修复组的牙周组织明显恢复。qRT-PCR结果显示,与对照组相比,LPS刺激组的骨膜增生蛋白和TGF-β1的表达量有所下降,但在LPS去除组则明显恢复:结论:随着炎症的发展,小鼠PDL中的骨膜素表达呈下降和上升趋势。结论:随着炎症的发展,小鼠 PDL 中的包膜生长因子表达呈下降和上升趋势,去除炎症刺激后包膜生长因子表达的明显恢复可能与 TGF-β1 有关,而 TGF-β1 对维持 PDL 的完整性至关重要。
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Temporal and spatial expression analysis of periostin in mice periodontitis model.

Objectives: This study aimed to investigate the temporal and spatial changes in the expression of periostin during periodontal inflammation in mice.

Methods: A periodontitis model was constructed using silk thread ligation. Mice were randomly divided into five groups including control group, 4-day ligation group, 7-day ligation group, 14-day ligation group, and self-healing group (thread removal for 14 days after 14-day ligation). Micro-CT and histological staining were performed to characterize the dynamic changes in the mouse periodontal tissue in each group. RNAscope and immunohistochemical staining were used to analyze the pattern of changes in periostin at various stages of periodontitis. The cell experiment was divided into three groups: control group, lipopolysaccharide (LPS) stimulation group (treated with LPS for 12 h), and LPS stimulation removal group (treated with LPS for 3 h followed by incubation with medium for 9 h). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of periostin, transforming growth factor-β1 (TGF-β1), and matrix metalloproteinase 2 (MMP2).

Results: Significant alveolar bone resorption was observed 7 days after ligation. With increasing duration of ligation, the damage to the mouse periodontal tissue was aggravated, which manifested as increased osteoclasts, widening of the periodontal membrane space, and decreased alveolar bone height. Some degree of periodontal tissue repair was observed in the self-healing group. Periostin expression decreased at 4 and 7 days compared with the control group and increased at 14 days compared with 4 and 7 days. A significant recovery was found in the self-healing group. The qRT-PCR results showed that the expression of periostin and TGF-β1 in the LPS stimulation group decreased compared with that in the control group but significantly recovered in the LPS removal group.

Conclusions: Periostin expression in the PDL of mice showed a downward and upward trend with inflammation progression. The significant recovery of periostin expression after removing inflammatory stimuli may be related to TGF-β1, which is crucial to maintain the integrity of the PDL.

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