Department of Veterinary Biosciences, Melbourne Veterinary School, The University of Melbourne, Parkville, Victoria, Australia.
Melbourne Dental School and The Bio21 Institute of Molecular Science and Biotechnology, The University of Melbourne, Parkville, Victoria, Australia.
J Periodontal Res. 2023 Jun;58(3):544-552. doi: 10.1111/jre.13120. Epub 2023 Mar 31.
Protease-activated receptor-2 (PAR ), a pro-inflammatory G-protein coupled receptor, has been associated with pathogenesis of periodontitis and the resulting bone loss caused by oral pathogens, including the keystone pathogen Porphyromonas gingivalis (P. gingivalis). We hypothesised that administration of a PAR antagonist, GB88, might prevent inflammation and subsequent alveolar bone resorption in a mouse model of periodontal disease.
Periodontitis was induced in mice by oral inoculations with P. gingivalis for a total of eight times over 24 days. The infected mice were treated with either GB88 or vehicle for the duration of the trial. Following euthanasia on day 56, serum was collected and used for the detection of mast cell tryptase. The right maxillae were defleshed and stained with methylene blue to measure the exposed cementum in molar teeth. The left maxillae were prepared for cryosections followed by staining for tartrate-resistant acid phosphatase to identify osteoclasts or with toluidine blue to identify mast cells. Reverse transcription quantitative PCR (RT-qPCR) was used to quantify the expression of inflammatory cytokines in the gingival tissue. Supernatants of T-lymphocyte cultures isolated from the regional lymph nodes were assayed using a cytometric bead array to measure the Th1/Th2/Th17 cytokine levels.
Measurement of the exposed cementum showed that GB88 reduced P. gingivalis-induced alveolar bone loss by up to 69%. GB88 also prevented the increase in osteoclast numbers observed in the infected mice. Serum tryptase levels were significantly elevated in both the infected groups, and not altered by treatment. RT-qPCR showed that GB88 prevented the upregulation of Il1b, Il6, Ifng and Cd11b. In T-lymphocyte supernatants, only IFNγ and IL-17A levels were increased in response to infection, but this was prevented by GB88 treatment.
GB88 significantly reduced osteoclastic alveolar bone loss in mice infected with P. gingivalis, seemingly by preventing the upregulation of several inflammatory cytokines. PAR antagonism may be an effective treatment strategy for periodontal disease.
蛋白酶激活受体 2（PAR ）是一种促炎 G 蛋白偶联受体，与牙周病的发病机制以及口腔病原体（包括关键病原体牙龈卟啉单胞菌（P. gingivalis））引起的骨丢失有关。我们假设，给予 PAR 拮抗剂 GB88 可能会预防牙周病小鼠模型中的炎症和随后的牙槽骨吸收。
通过口腔接种 P. gingivalis 总共 8 次，在 24 天内诱导牙周炎。在整个试验过程中，用 GB88 或载体处理感染的小鼠。在第 56 天安乐死后，收集血清并用于检测肥大细胞胰蛋白酶。去除右侧上颌骨的肉，用亚甲蓝染色以测量磨牙中暴露的牙骨质。准备左侧上颌骨进行冷冻切片，然后用抗酒石酸酸性磷酸酶染色以识别破骨细胞，或用甲苯胺蓝染色以识别肥大细胞。使用逆转录定量 PCR（RT-qPCR）定量牙龈组织中炎症细胞因子的表达。使用细胞因子珠阵列测定从局部淋巴结分离的 T 淋巴细胞培养物的上清液，以测量 Th1/Th2/Th17 细胞因子水平。
暴露牙骨质的测量表明，GB88 可使 P. gingivalis 诱导的牙槽骨丢失减少多达 69%。GB88 还防止了感染小鼠中观察到的破骨细胞数量的增加。感染组的血清胰蛋白酶水平均显着升高，而治疗并未改变。RT-qPCR 显示，GB88 可预防 Il1b、Il6、Ifng 和 Cd11b 的上调。在 T 淋巴细胞上清液中，仅 IFNγ 和 IL-17A 水平因感染而增加，但这可通过 GB88 治疗来预防。
GB88 可显着减少感染 P. gingivalis 的小鼠的破骨细胞性牙槽骨丢失，似乎是通过防止几种炎症细胞因子的上调来实现的。PAR 拮抗可能是牙周病的有效治疗策略。
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