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抗菌光动力疗法对烧伤患者创面感染分离的金黄色葡萄球菌生物膜相关基因表达的影响。

The effect of antimicrobial photodynamic therapy on the expression of biofilm associated genes in Staphylococcus aureus strains isolated from wound infections in burn patients.

机构信息

Department of Microbiology, Hamadan University of Medical Sciences, Hamadan, Iran.

Dental Research Center, Dentistry Research Institute, Tehran University of Medical Sciences, Tehran, Iran.

出版信息

Photodiagnosis Photodyn Ther. 2019 Mar;25:406-413. doi: 10.1016/j.pdpdt.2019.01.028. Epub 2019 Jan 23.

Abstract

PURPOSE

Burn patients are particularly susceptible to microbial infection. Staphylococcus aureus causes burn wound, impetigo and cellulitis. Although sub-lethal antimicrobial photodynamic therapy (aPDT) would not result in microorganism killing, it can considerably influence microbial virulence factor.

METHODS

Twelve methicillin-resistant S. aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) isolated from burns patients. To determine the sub-lethal dose of aPDT, 12 clinical isolates of S. aureus photosensitized with 100 μg ml toluidine blue O (TBO) and irradiated by light emitting diode (LED) with a wavelength of 630 ± 10 nm and energy densities of 52.0, 104.1, and 156.2 J/cm, then bacterial viability was measured. The effects of sub-lethal aPDT on the expression levels of ica ABCD and ica R genes were assessed by quantitative Real-time PCR (qRT-PCR) method.

RESULT

Fifty and 100 μg ml of TBO significantly reduced the mean cell survival in the MRSA (2.5 - 3 log) and MSSA (2.75-3.1 log) isolates. The average expression levels of icaA, ica B, ica C, and ica D in the MRSA and MSSA isolates were decreased by (12, 14, 11, and 9) and (13, 14.5, 12, and 9.5) fold change, respectively (P < 0.05). However, the expression of ica R gene was decreased by 6 and 8 folds change in MRSA and MSSA, respectively.

CONCLUSION

The potential of TBO-mediated aPDT could reduce the expression of ica ABCD as important genes involved in biofilm formation and ica R gene as a repressor of the ica operon. Therefore, the use of aPDT agents as a complementary therapy in wound infections of burn patients is recommended.

摘要

目的

烧伤患者特别容易受到微生物感染。金黄色葡萄球菌会导致烧伤创面、脓疱病和蜂窝织炎。虽然亚致死剂量的光动力抗菌疗法(aPDT)不会导致微生物死亡,但它可以显著影响微生物毒力因子。

方法

从烧伤患者中分离出 12 株耐甲氧西林金黄色葡萄球菌(MRSA)和甲氧西林敏感金黄色葡萄球菌(MSSA)。为了确定 aPDT 的亚致死剂量,用 100μg/ml 的甲苯胺蓝 O(TBO)对 12 株金黄色葡萄球菌进行敏化,然后用波长为 630±10nm 和能量密度为 52.0、104.1 和 156.2J/cm 的发光二极管(LED)照射,然后测量细菌的存活率。通过实时荧光定量 PCR(qRT-PCR)方法评估亚致死 aPDT 对 icaABCD 和 icaR 基因表达水平的影响。

结果

50μg/ml 和 100μg/ml 的 TBO 显著降低了 MRSA(2.5-3 对数)和 MSSA(2.75-3.1 对数)分离株的平均细胞存活率。MRSA 和 MSSA 分离株中 icaA、icaB、icaC 和 icaD 的平均表达水平分别降低了 12、14、11 和 9 倍和 13、14.5、12 和 9.5 倍(P<0.05)。然而,MRSA 和 MSSA 的 icaR 基因表达水平分别降低了 6 和 8 倍。

结论

TBO 介导的 aPDT 可能降低与生物膜形成有关的重要基因 icaABCD 的表达和 ica 操纵子的抑制剂 icaR 基因的表达。因此,建议将 aPDT 剂作为烧伤患者伤口感染的辅助治疗。

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