目的：探讨牙龈卟啉单胞菌（Pg）持续存在（Ps）对巨噬细胞免疫炎症反应的影响，并探讨其潜在机制。方法:将Pg细胞培养至稳定期(72 h),随后用高浓度甲硝唑100 mg/L、阿莫西林100 mg/L及其联合处理不同时间段,命名为甲硝唑组,阿莫西林组和（甲硝唑阿莫西林）组。将未经管理的Pg细胞作为空白对照。通过集落形成单位测定来测量 PgPs 细胞的存活情况。通过Live/Dead染色观察PgPs的活状态。然后，用Pg和甲硝唑处理的PgPs（M-PgPs）处理巨噬细胞，命名为Pg组和M-PgPs组。使用透射电子显微镜（TEM）观察巨噬细胞中的细菌。通过实时荧光定量PCR和酶联免疫吸附试验测定巨噬细胞中促炎细胞因子的表达水平。通过共聚焦免疫荧光显微镜检测叉头盒 1 (FOXO1) 的位置。分别用抑制剂（Fi）或激活剂（Fa）抑制或增强FOXO1表达后，用Pg和M-PgPs处理巨噬细胞，分为空白组、Pg组、M-PgPs组、Fi组，（Fi Pg）组、(Fi M-PgPs)组、Fa组、(Fa Pg)组和(Fa M-PgPs)组。然后，评估促炎细胞因子的表达模式。结果：在浮游培养物和用甲硝唑、阿莫西林或两者处理的 Pg 生物膜中观察到大量存活的 PgP，并且这些持续者可以形成新的菌落。 Pg和M-PgPs能够进入巨噬细胞，白细胞介素(IL)-1β、IL-6、IL-8和肿瘤坏死因子-α(TNF-α)的蛋白表达水平[Pg组：(2 392 ±188)、(162±29)、(5 558±661)、(789±155)μg/L； M-PgPs组：(2 415±420)、(155±3)、(5 732±782)、(821±176) μg/L)]较空白组显着上调[(485±140)、 （21±9）、（2 332±87）、（77±7）μg/L]（P<0.01）。此外，Pg和M-PgPs可以促进FOXO1的核转位和积累。此外，与空白组相比，FOXO1、BCL6和KLF2 mRNA相对表达水平上调（P<0.05）。 Fi Pg组IL-1β、IL-6、IL-8、TNF-α蛋白表达量[(1 081±168)、(70±8)、(1 976±544)、(420± 47) μg/L]显着低于Pg组[(4 411±137)、(179±6)、(5 161±929)、(934±24) μg/L](P<0.05)。同样，Fi M-PgPs组IL-1β、IL-6、IL-8、TNF-α蛋白表达量[(1 032±237)、(74±10)、(1 861±614)、( 405±32) μg/L]显着低于M-PgPs组[(4 342±314)、(164±17)、(4 438±1 374)、(957±25) μg/L](P< 0.05）。 Fa Pg组IL-1β、IL-6、IL-8、TNF-α蛋白表达量[(8 198±1 825)、(431±28)、(8 919±650)、 (2 186±301) μg/L]和Fa M-PgPs组[(8 159±2 627)、(475±26)、(8 995±653)、(2 255±387 μg/L)均显着升高。分别高于Pg组和M-PgPs组(P<0.05)。结论：PgPs 对甲硝唑和阿莫西林高度耐受。 M-PgPs可以通过上调FOXO1信号通路来增强巨噬细胞的免疫炎症反应，而这种作用与Pg没有显着差异。

Objective: To investigate the effects of Porphyromonas gingivalis (Pg) persisters (Ps) on immuno-inflammatory responses in macrophages, and to explore the underlying mechanisms. Methods: Pg cells were cultured to the stationary phase (72 h), and subsequently treated by high concentration of metronidazole at 100 mg/L, amoxicillin at 100 mg/L and the combination of them for different time period, named as metronidazole group, amoxicillin group and (metronidazole+amoxicillin) group. Pg cells without management were used as blank control. The survival profile of PgPs cells was measured by colony-forming unit assay. The living state of PgPs was observed by Live/Dead staining. Then, Pg and metronidazole-treated PgPs (M-PgPs) were used to treat macrophages, named as Pg group and M-PgPs group. Transmission electron microscopy (TEM) was used to observe the bacteria in the macrophages. The expression levels of proinflammatory cytokines in macrophages were determined by real-time fluorescence quantitative PCR and enzyme-linked immunosorbent assay. The location of forkhead box 1 (FOXO1) was detected by confocal immunofluorescence microscopy. After inhibiting or enhancing the FOXO1 expressions using inhibitors (Fi) or activators (Fa) respectively, the macrophages were treated with Pg and M-PgPs, divided as Blank group, Pg group, M-PgPs group, Fi group, (Fi+Pg) group, (Fi+M-PgPs) group, Fa group, (Fa+Pg) group and (Fa+M-PgPs) group. Then, the expression pattens of proinflammatory cytokines were assessed. Results: Remarkable number of lived PgPs was observed, both in planktonic culture and Pg biofilms either treated with metronidazole, amoxicillin or both, and those persisters could form new colonies. Pg and M-PgPs were able to enter into the macrophages and the protein expression levels of interleukin (IL)-1β, IL-6, IL-8 and tumor necrosis factor-α (TNF-α) [Pg group: (2 392±188), (162±29), (5 558±661), (789±155) μg/L; M-PgPs group: (2 415±420), (155±3), (5 732±782), (821±176) μg/L)] were significantly upregulated than those in Blank group [(485±140), (21±9), (2 332±87), (77±7) μg/L] (P<0.01). Moreover, Pg and M-PgPs could facilitate the nuclear translocation and accumulation of FOXO1. In addition, the relative mRNA expression levels of FOXO1, BCL6 and KLF2 were upregulated when compared to Blank group (P<0.05). Furthermore, the protein expression levels of IL-1β, IL-6, IL-8 and TNF-α in Fi+Pg group [(1 081±168), (70±8), (1 976±544), (420±47) μg/L] were remarkably lower than Pg group [(4 411±137), (179±6), (5 161±929), (934±24) μg/L] (P<0.05). Similarly, the protein expression levels of IL-1β, IL-6, IL-8 and TNF-α in Fi+M-PgPs group [(1 032±237), (74±10), (1 861±614), (405±32) μg/L] were remarkably lower than M-PgPs group [(4 342±314), (164±17), (4 438±1 374), (957±25) μg/L] (P<0.05). On the contrary, the protein expression levels of IL-1β, IL-6, IL-8 and TNF-α in Fa+Pg group [(8 198±1 825), (431±28), (8 919±650), (2 186±301) μg/L] and Fa+M-PgPs group [(8 159±2 627), (475±26), (8 995±653), (2 255±387 μg/L) were both significantly higher than Pg group and M-PgPs group, respectively (P<0.05). Conclusions: PgPs are highly tolerant to metronidazole and amoxicillin. The M-PgPs could enhance the immuno-inflammatory responses in macrophages by upregulating the FOXO1 signaling pathway, while this effect exhibits no significant difference with Pg.