Macrophage M1 polarization mediated via the IL-6/STAT3 pathway contributes to apical periodontitis induced by Porphyromonas gingivalis

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Chen, Xuan; Dou, Jinge; Fu, Zhuohui; Qiu, Yang; Zou, Ling; Huang, Dingming; Tan, Xuelian, 2022, "Macrophage M1 polarization mediated via the IL-6/STAT3 pathway contributes to apical periodontitis induced by Porphyromonas gingivalis", https://doi.org/10.48331/scielodata.UNJC4K, SciELO Data, V1, UNF:6:XYVn3HLR+CX4EJfvpKpN9w== [fileUNF]
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Description
Objective: To investigate the involvement of IL-6/STAT3 signaling pathway activation in macrophage polarization and bone destruction related to apical periodontitis (AP) stimulated by Porphyromonas gingivalis. Methodology: Macrophage polarization, IL-6/STAT3 expression, and the presence of P. gingivalis were detected in human AP tissues via RT-qPCR, western blotting, and immunohistochemistry staining. Murine bone marrow derived macrophages were isolated and cultured with P. gingivalis W83 in vitro, and levels of macrophage IL-6 expression, STAT3 phosphorylation, and macrophage polarization with or without the selective STAT3 phosphorylation inhibitor Stattic (5 μM) were detected via ELISA, western blotting, RT-qPCR, and flow cytometry, respectively. P. gingivalis-induced murine AP models were constructed, and bone destruction and macrophage polarization in the apical region were evaluated. Transwell co-culture systems were used to investigate the effects of macrophages infected with P. gingivalis on osteogenesis and osteoclastogenesis. Results: P. gingivalis was detected in human AP tissues that highly expressed IL-6/STAT3, and the M1 subtype of macrophages was more abundant in these tissues. P. gingivalis infection induced IL-6 expression, STAT3 phosphorylation, and M1 polarization of macrophages, while 5 μM of Stattic partially abolished these activation effects. Systemic STAT3 blockade via oral administration of Stattic at a dose of 25 mg kg-1 alleviated murine periapical bone resorption and apical infiltration of M1 macrophages induced by P. gingivalis infection in vivo. Furthermore, macrophages infected with P. gingivalis promoted bone destruction via secretion of IL-6, TNF-α, and RANKL, which hinder pre-osteoblast expression of Runx2 and accelerate pre-osteoclast expression of NFAT2. Conclusions: The activation of IL-6/STAT3 signaling pathway is involved in mediating macrophages M1 polarization in the P. gingivalis induced apical inflammatory context and may also be intimately involved in the bone loss caused by P. gingivalis infection, directing the M1 macrophage infiltration during the progression of AP.
Subject Medicine, Health and Life Sciences
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fig S3A-no-stattic.tif
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Published Nov 3, 2022
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MD5: cd54ce3027ac8d3d3f5a0d20f1548c8b
Fig S3. Macrophage morphology after 12h stimulation of 0um of Stattic.
fig S3C-7.5um-stattic.tif
TIFF Image - 4.7 MB
Published Nov 3, 2022
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MD5: 951c7a5dd174cb2fe253f5bdedec4e86
Fig S3. Macrophage morphology after 12h stimulation of 7.5um of Stattic.
Fig-1-MFI-data.csv
Comma Separated Values - 320 B
Published Nov 3, 2022
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MD5: 9b4f6227579eb491256475e5a9611e91
The estimated mean fluorescence intensities in figure 1.
FIG-2b-data-for-the-WB-gray-scale.tab
Tabular Data - 108 B
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2 Variables, 6 Observations UNF:6:3ItJLPkFg/WNSjHgePE/Ng==
The estimated gray scales for the WB band in figure 2.
Fig-2b-IL-6-expression+pg-qpcr.csv
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Fig 2B. The q-PCR results of IL-6 expression and P. gingivalis presence in relative to β-actin in human samples.
Fig-3-IL-6-stat3-QPCR.csv
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MD5: 9031ee4cf4f3d364d5477a987c724e65
Fig. 3. The q-PCR results of IL-6 and Stat3 expression in relative to β-actin in P. gingivalis stimulated macrophages.
Fig-3E-data-for-the-WB-gray-scale.tab
Tabular Data - 60 B
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2 Variables, 6 Observations UNF:6:7h++YDzu+82clFafjuwnGw==
The estimated gray scales for the WB band in figure 3E.
Fig-4-IL-1beta-TNF-alpha-qpcr.csv
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Fig 4. The q-PCR results of IL-1β and TNF-α expression in relative to β-actin in P. gingivalis stimulated macrophages.
Fig-4-IL-6-ELISA-no-stattic.csv
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Fig 4. The ELISA results of IL-6 secretion in P. gingivalis stimulated macrophages without the stimulation of Stattic.
Fig-4-IL-6-ELISA-with-stattic.csv
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Fig 4. The ELISA results of IL-6 secretion in P. gingivalis stimulated macrophages with the stimulation of Stattic.
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Contact Xuelian TAN and Dingming HUANG - tanxuelian@scu.edu.cn; dingminghuang@163.com
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