背景:塞来昔布,一种抗炎药,正在研究使用抗微生物剂和免疫调节剂药物的联合疗法。
目的:评估塞来昔布是否对巴西副球菌具有直接的体外抗真菌作用,副病菌(PCM)的病原体,如果它提高了中性粒细胞(PMN)在该疾病的实验性小鼠皮下(气囊)模型中的体内活性。
方法:使用微量稀释技术评估了塞来昔布(6mg/mL)对巴西假单胞菌(Pb18)的抗真菌活性。将与Pb18共培养并用塞来昔布(6mg/mL)处理的脾细胞共培养24、48和72小时。用Pb18接种瑞士小鼠,并在皮下气囊中用塞来考昔(6mg/kg)处理。从气囊收集嗜中性粒细胞。线粒体活性,活性氧生产,过氧化氢酶,过氧化物酶,细胞因子和趋化因子,氮物种,总蛋白质,分析了PMN的杀微生物活性和活的Pb18细胞数量。
结果:塞来昔布对与Pb18共培养的脾细胞没有细胞毒性作用,但具有明显的直接抗真菌作用,在体外和体内抑制真菌生长。塞来昔布与气囊中的免疫系统细胞相互作用,它导致PMN的激活,如几个参数(线粒体活性,活性氧,过氧化物酶,KC和IL-6增加,杀伤常数和吞噬作用)。塞来昔布能够减少IL-4、IL-10和IL-12细胞因子的产生。回收的活Pb18的数量急剧下降。
结论:这是关于塞来昔布对巴西假单胞菌的直接抗真菌活性的首次报道。塞来昔布的使用为PCM的未来治疗开辟了新的可能性。
BACKGROUND: Celecoxib, an anti-inflammatory drug, combined therapies using antimicrobials and immune modulator drugs are being studied.
OBJECTIVE: To assess whether Celecoxib has direct in vitro antifungal effect against the Paracoccidioides brasiliensis, the causative agent of Paracoccidioidomycosis-(PCM) and also if it improves the in vivo activity of neutrophils-(PMN) in an experimental murine subcutaneous-(air pouch) model of the disease.
METHODS: The antifungal activity of Celecoxib(6 mg/mL) on P. brasiliensis-(Pb18) was evaluated using the microdilution technique. Splenocytes co-cultured with Pb18 and treated with Celecoxib(6 mg/mL) were co-cultured for 24, 48 and 72-hours. Swiss mice were inoculated with Pb18 and treated with Celecoxib(6 mg/kg) in the subcutaneous air pouch. Neutrophils were collected from the air pouch. Mitochondrial activity, reactive oxygen production, catalase, peroxidase, cytokines and chemokines, nitrogen species, total protein, microbicidal activity of PMNs and viable Pb18 cells numbers were analyzed.
RESULTS: Celecoxib had no cytotoxic effect on splenocytes co-cultured with Pb18, but had a marked direct antifungal effect, inhibiting fungal growth both in vitro and in vivo. Celecoxib interaction with immune system cells in the air pouch, it leads to activation of PMNs, as confirmed by several parameters (mitochondrial activity, reactive oxygen species, peroxidase, KC and IL-6 increase, killing constant and phagocytosis). Celecoxib was able to reduce IL-4, IL-10 and IL-12 cytokine production. The number of recovered viable Pb18 decreased dramatically.
CONCLUSIONS: This is the first report of the direct antifungal activity of Celecoxib against P. brasiliensis. The use of Celecoxib opens a new possibility for future treatment of PCM.