Li Gege, Pan Jiahui, Tang Qiuling, Liu Xinchan, Wang Liuran, Meng Yang, Yu Weixian
Department of Periodontics, School of Stomatology, Jilin University, Changchun, Jilin, China.
Iran J Immunol. 2018 Jun;15(2):84-96.
C5a receptor antagonist PMX205 is a synthetic hexapeptide capable of blocking C5a-C5a receptor (C5aR) axis by simulating C5a active C-terminal amino acid residues. This hexapeptide presents good anti-inflammatory effects in a series of inflammation models. The anti-inflammatory effect of PMX205 on periodontitis is yet to be fully fathomed.
To examine the anti-inflammatory effects of PMX205 on RAW264.7 mouse macrophages exposed to gingipain extracts and Porphyromonas gingivalis (P. gingivalis).
MTT assay was carried out so as to specify the cytotoxicity of PMX205. RAW264.7 cells were co-cultured in vitro with gingipain extracts or P. gingivalis to simulate the periodontitis inflammatory milieu. Real-time quantitative PCR, ELISA and Griess assay were performed in order to detect tumor necrosis factor-α (TNF-α), IL-6, IL-23, nitric oxide (NO), IL-10, transforming growth factor-β1 (TGF-β1), andarginase-1 (Arg-1). Furthermore, phagocytosis assay was done to evaluate the phagocytic capacity of RAW 264.7 cells. Finally, western blot analysis was conducted to evaluate myeloid differentiation factor 88 (MyD88).
PMX205 increased the expression levels of bacteriostatic substances (NO and IL-23) and anti-inflammatory cytokines (TGF-β1, IL-10 and Arg-1); however, it reduced the expression levels of proinflammatory cytokines TNF-α and IL-6 once RAW 264.7 macrophages were stimulated via gingipain extracts or P. gingivalis. In addition, PMX205 promoted the macrophage phagocytosis and down-regulated protein expression of MyD88.
PMX205 has recognizable anti-inflammatory effects in RAW 264.7 cell inflammation model, a finding which probably opens doors to future investigations on new targets for the prevention and treatment of chronic periodontitis.
C5a受体拮抗剂PMX205是一种合成六肽，能够通过模拟C5a活性C末端氨基酸残基来阻断C5a - C5a受体（C5aR）轴。这种六肽在一系列炎症模型中呈现出良好的抗炎作用。PMX205对牙周炎的抗炎作用尚未完全明了。
研究PMX205对暴露于牙龈蛋白酶提取物和牙龈卟啉单胞菌的RAW264.7小鼠巨噬细胞的抗炎作用。
进行MTT试验以确定PMX205的细胞毒性。RAW264.7细胞与牙龈蛋白酶提取物或牙龈卟啉单胞菌在体外共培养，以模拟牙周炎炎症环境。进行实时定量PCR、ELISA和Griess试验，以检测肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、白细胞介素-23（IL-23）、一氧化氮（NO）、白细胞介素-10（IL-10）、转化生长因子-β1（TGF-β1）和精氨酸酶-1（Arg-1）。此外，进行吞噬试验以评估RAW 264.7细胞的吞噬能力。最后，进行蛋白质印迹分析以评估髓样分化因子88（MyD88）。
PMX205增加了抑菌物质（NO和IL-23）和抗炎细胞因子（TGF-β1、IL-10和Arg-1）的表达水平；然而，当RAW 264.7巨噬细胞通过牙龈蛋白酶提取物或牙龈卟啉单胞菌刺激时，它降低了促炎细胞因子TNF-α和IL-6的表达水平。此外，PMX205促进了巨噬细胞吞噬作用，并下调了MyD88的蛋白表达。
PMX205在RAW 264.7细胞炎症模型中具有明显的抗炎作用，这一发现可能为未来慢性牙周炎预防和治疗新靶点的研究开辟道路。
