背景:本研究的目的是从临床分离的铜绿假单胞菌中提取绿脓苷,表征其化学性质,并评估其对不同细菌和癌细胞的生物活性。由于其不同的生物活性特性,绿脓苷,作为铜绿假单胞菌的毒力因子之一,持有一个有希望的,安全,和可用的治疗潜力。
方法:收集来自不同感染源的30株临床铜绿假单胞菌,并通过常规方法进行鉴定。VITEK2紧凑型系统,和16SrRNA。吩嗪修饰基因(phzM,phzS)使用聚合酶链反应(PCR)鉴定。花青素的化学表征包括紫外-可见分光光度法,傅里叶变换红外光谱(FTIR),气相色谱-质谱(GC-MS),和液相色谱-质谱(LC-MS)。通过确定针对不同临床细菌菌株的MIC值并使用细胞毒性评估其针对A549,MDA-MB-231和Caco-2癌细胞系的抗癌活性来探索绿脓苷的生物学活性,伤口愈合和菌落形成试验。
结果:所有鉴定的分离株都含有至少一个phzM或phzS基因。在13个分离株中证明了这两种基因的共存。UV-VIS吸收峰在215、265、385和520nm处最大。FTIR可以识别特征的绿脓氰素官能团,而GC-MS和LC-MS都阐明了化学式C11H18N2O2,分子量为210。四技术分析方法证实了提取的绿脓苷的化学性质。提取物具有广谱抗菌活性,对芽孢杆菌的活性最大,葡萄球菌,和链球菌(MIC31.25-125µg/mL),其次是大肠杆菌分离株(MICs250-1000µg/mL)。关于抗癌活性,绿脓苷提取物对A549、MDA-MB-231和Caco-2癌细胞系的IC50值为130、105和187.9µg/mL,分别。此外,在这些细胞中,花青素显着抑制了集落形成和迁移能力。
结论:已证明提取的绿脓苷是针对各种细菌感染和癌症的潜在有效候选物。因此,目前的发现可能有助于通过负担得起的方法轻松生产这种天然化合物。尽管如此,未来的研究需要研究在体内的药效,并分析其与其他传统抗生素或抗癌药物联合使用的结果。
BACKGROUND: The objectives of the current study were to extract
pyocyanin from Pseudomonas aeruginosa clinical isolates, characterize its chemical nature, and assess its biological activity against different bacteria and cancer cells. Due to its diverse bioactive properties,
pyocyanin, being one of the virulence factors of P. aeruginosa, holds a promising, safe, and available therapeutic potential.
METHODS: 30 clinical P. aeruginosa isolates were collected from different sources of infections and identified by routine methods, the VITEK 2 compact system, and 16 S rRNA. The phenazine-modifying genes (phzM, phzS) were identified using polymerase chain reaction (PCR). Pyocyanin chemical characterization included UV-Vis spectrophotometry, Fourier Transform Infra-Red spectroscopy (FTIR), Gas Chromatography-Mass Spectrometry (GC-MS), and Liquid Chromatography-Mass Spectrometry (LC-MS). The biological activity of pyocyanin was explored by determining the MIC values against different clinical bacterial strains and assessing its anticancer activity against A549, MDA-MB-231, and Caco-2 cancer cell lines using cytotoxicity, wound healing and colony forming assays.
RESULTS: All identified isolates harboured at least one of the phzM or phzS genes. The co-presence of both genes was demonstrated in 13 isolates. The UV-VIS absorbance peaks were maxima at 215, 265, 385, and 520 nm. FTIR could identify the characteristic pyocyanin functional groups, whereas both GC-MS and LC-MS elucidated the chemical formula C11H18N2O2, with a molecular weight 210. The quadri-technical analytical approaches confirmed the chemical nature of the extracted
pyocyanin. The extract showed broad-spectrum antibacterial activity, with the greatest activity against Bacillus, Staphylococcus, and Streptococcus species (MICs 31.25-125 µg/mL), followed by E. coli isolates (MICs 250-1000 µg/mL). Regarding the anticancer activity, the
pyocyanin extract showed IC50 values against A549, MDA-MB-231, and Caco-2 cancer cell lines of 130, 105, and 187.9 µg/mL, respectively. Furthermore, pyocyanin has markedly suppressed colony formation and migratory abilities in these cells.
CONCLUSIONS: The extracted pyocyanin has demonstrated to be a potentially effective candidate against various bacterial infections and cancers. Hence, the current findings could contribute to producing this natural compound easily through an affordable method. Nonetheless, future studies are required to investigate
pyocyanin\'s effects in vivo and analyse the results of combining it with other traditional antibiotics or anticancer drugs.