| 俞 梅,李 娜,喻锦娴,王文茜,刘思佳,徐 佳.牙龈卟啉单胞菌脂多糖通过TXNIP/Nlrp3炎性通路对小鼠牙周膜成纤维细胞迁移的影响[J].,2022,(5):832-836 |
| 牙龈卟啉单胞菌脂多糖通过TXNIP/Nlrp3炎性通路对小鼠牙周膜成纤维细胞迁移的影响 |
| Effect of Porphyromonas Gingivalis Lipopolysaccharide on Migration of Periodontal Ligament Fibroblasts ViaTXNIP/Nlrp3 Inflammasome Pathway |
| 投稿时间：2021-07-03 修订日期：2021-07-27 |
| DOI：10.13241/j.cnki.pmb.2022.05.007 |
| 中文关键词: 牙龈卟啉单胞菌脂多糖 TXNIP/Nlrp3途径 牙周膜成纤维细胞 迁移 |
| 英文关键词: Lipopolysaccharide from Porphyromonas gingivalis TXNIP/Nlrp3 pathway Periodontal ligament fibroblast Migration |
| 基金项目:国家卫生计生委医药卫生科技发展研究中心项目（W2015CAE173） |
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| 中文摘要: |
| 摘要 目的：研究牙龈卟啉单胞菌脂多糖（Porphyromonas lipopolysaccharide，LPS-PG）对牙周膜成纤维细胞（mouse periodontal Ligament: Normal Fibroblasts，mPDLFs）增殖及迁移的影响，探讨TXNIP/Nlrp3炎性体途径在其中的作用。方法：采用不同浓度的LPS-PG刺激小鼠mPDLFs细胞不同时间，CCK-8法检测细胞增殖抑制率。然后将细胞分为对照组（培养基作用24 h）和LPS-PG组（2 M的LPS-PG作用24 h），划痕实验检测细胞迁移，ELISA法检测白细胞介素-1β（Interleukin-1β，IL-1β）和高迁移率族蛋白B1（High mobility group protein B1，HMGB1）的水平，Western Blot检测Nod样受体蛋白3（Nod-like receptor pyrin domain3，NLRP3）、凋亡相关斑点样蛋白（Apoptosis-associated speck-like protein，ASC）、活化半胱氨酸蛋白酶（cleaved-caspase-1）和硫氧还蛋白相互作用蛋白（Thioredoxin-interacting protein,TXNIP）的表达，免疫共沉淀检测以上蛋白间的相互作用。结果：与0 M的LPS-PG相比，1 M和2 M的LPS-PG可显著增加mPDLFs的细胞增殖抑制率（P<0.05）。与LPS-PG作用0 h相比，LPS-PG作用12 h、24 h和48 h均可显著增加mPDLFs的细胞增殖抑制率（P<0.05）。LPS-PG组细胞向中间'伤口'迁移的距离及细胞数量远远低于对照组。与对照组相比，LPS-PG组细胞中IL-1β和HMGB1的水平、NLRP3、cleaved-caspase-1和TXNIP蛋白表达均显著增加，且NLRP3和ASC、NLRP3和cleaved-caspase-1、TXNIP和NLRP3的蛋白互作能力显著增强（P<0.05），而两组ASC蛋白的表达无显著差异（P＞0.05）。结论：LPS-PG可抑制mPDLFs细胞的增殖和迁移，其机制与Nlrp3炎性体的形成与活化有密切的关系。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effects of lipopolysaccharide from Porphyromonas gingivalis (LPS-PG) on the proliferation and migration of periodontal ligament fibroblasts (mPDLFs), and to explore the role of TXNIP/Nlrp3 inflammasome pathway in it. Methods: Different concentrations of LPS-PG were used to stimulate mouse mPDLFs cells for different times , and the cell proliferation inhibition rate was detected by CCK-8 method. The cells were then divided into two groups: control group (stimulation with medium for 24 h) and LPS-PG group (stimulation with 2 M LPS-PG for 24 h). The scratch test was used to detect cell migration, ELISA method was used to detect the level of IL-1β and HMGB1, Western Blot was used to detect the expression of NLRP3, ASC, cleaved-caspase-1 and TXNIP protein, and Co-immunoprecipitation is used to detect the interaction between the above proteins. Results: Compared with 0 M of LPS-PG, 1 M and 2 M of LPS-PG significantly increased the cell proliferation inhibition rate of mPDLFs(P<0.05). Compared with LPS-PG at 0 h, LPS-PG at 12 h, 24 h and 48 h can significantly increase the cell proliferation inhibition rate of mPDLFs(P<0.05). The migration distance and the number of cells in the LPS-PG group to the middle 'wound' were much lower than those in the control group. Compared with the control group, the levels of IL-1β and HMGB1 and the expression of NLRP3, cleaved-caspase-1 and TXNIP protein in the LPS-PG group were significantly increased(P<0.05), and the protein interaction ability of NLRP3 and ASC, NLRP3 and cleaved-caspase-1, TXNIP and NLRP3 was significantly enhanced(P<0.05), but there was no significant difference in the expression of ASC protein between the two groups(P>0.05). Conclusion: LPS-PG can inhibit the proliferation and migration of mPDLFs cells, and its mechanism is closely related to the formation and activation of Nlrp3 inflammasome. |
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