背景:虽然传统玛雅医学中使用的许多植物对传染病的抗菌活性已被证明,它们抑制群体感应(QS)作为发现新型抗毒剂的手段的潜力仍未开发。
目的:通过测定传统玛雅药物中植物对铜绿假单胞菌QS调节毒力因子的抑制作用,评价其抗毒力潜力。
方法:选择一组用于传统玛雅医学抗传染病的植物,使用参考菌株铜绿假单胞菌PA14WT评估其甲醇提取物在10mg/mL下的抗菌和抗毒活性。肉汤微量稀释法测定抗菌活性(MIC),同时通过测量抗生物膜作用以及对绿脓苷和蛋白酶活性的抑制作用来评估其抗毒力活性。使用液-液分配程序分离最具生物活性的提取物,并以5mg/mL评估半纯化的级分的抗菌和抗毒活性。
结果:选择了17种传统上用于治疗感染相关疾病的玛雅药用植物。提取物均未表现出抗菌活性,而抗毒力活性在黄麻提取物中检测到,BurseraSimaruba,Caprariabiflora,仙叶,帕雷拉香波罗斯和尤卡坦纳斯。对生物膜形成最活跃的提取物(74%和69%的抑制作用)来自C.esculifolia(树皮)和Yucatanensis(根),分别。或者,B.flammea(根)的提取物,B.simaruba(树皮),C.帕雷拉(根),和C.biflora(根),减少了绿脓苷和蛋白酶的产生(50-84%和30-58%,分别)。通过对念珠菌的生物活性根提取物进行分级分离,可以鉴定出两种具有抗毒力活性的半纯化级分。
结论:在火焰芽孢杆菌的粗提物中检测到的抗毒力活性,B.simaruba,C.Biflora,C.七叶,C.帕雷拉,和C.Yucatanensis,证实了这些药用植物对传染病的功效和传统用途。该提取物和半纯化的热杆菌级分的活性表明在铜绿假单胞菌中存在能够干扰QS的亲水性代谢物。这项研究代表了具有抗QS特性的玛雅药用植物的第一份报告,并表明它们代表了新型抗毒剂的重要来源。
BACKGROUND: While the antimicrobial activity of a number of plants used in traditional Mayan medicine against infectious diseases has been documented, their potential to inhibit quorum sensing (QS) as means of discovering novel anti-virulence agents remains unexplored.
OBJECTIVE: To evaluate the anti-virulence potential of plants used in traditional Mayan medicine by determining their inhibition of QS- regulated virulence factors in Pseudomonas aeruginosa.
METHODS: A group of plants used in traditional Mayan medicine against infectious diseases was selected, and their methanolic extracts were evaluated at 10 mg/mL for their antibacterial and anti-virulence activity using the reference strain P. aeruginosa PA14WT. The broth microdilution method was used to determine antibacterial activity (MIC), while anti-virulence activity was evaluated by measuring the anti-biofilm effect and the inhibition of pyocyanin and protease activities. The most bioactive extract was fractionated using a liquid-liquid partition procedure and the semipurified fractions were evaluated at 5 mg/mL for antibacterial and anti-virulence activity.
RESULTS: Seventeen Mayan medicinal plants traditionally used to treat infection-associated diseases were selected. None of the extracts exhibited antibacterial activity, whereas anti-virulence activity was detected in extracts of Bonellia flammea, Bursera simaruba, Capraria biflora, Ceiba aesculifolia, Cissampelos pareira and Colubrina yucatanensis. The most active extracts (74% and 69% inhibition) against biofilm formation were from C. aesculifolia (bark) and C. yucatanensis (root), respectively. Alternatively, the extracts of B. flammea (root), B. simaruba (bark), C. pareira (root), and C. biflora (root), reduced pyocyanin and protease production (50-84% and 30-58%, respectively). Fractionation of the bioactive root extract of C. yucatanensis allowed the identification of two semipurified fractions with anti-virulence activity.
CONCLUSIONS: The anti-virulence activity detected in the crude extracts of B. flammea, B. simaruba, C. biflora, C. aesculifolia, C. pareira, and C. yucatanensis, confirms the efficacy and traditional use of these medicinal plants against infectious diseases. The activity of the extract and semipurified fractions of C. yucatanensis indicates the presence of hydrophilic metabolites capable of interfering with QS in P. aeruginosa. This study represents the first report of Mayan medicinal plants with anti-QS properties and suggests they represent an important source of novel anti-virulence agents.