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临床牙龈卟啉单胞菌分离株中精氨酸A特异性半胱氨酸蛋白酶基因表达谱针对光活化消毒光杀伤作用的研究。

Investigation of arginine A-specific cysteine proteinase gene expression profiling in clinical Porphyromonas gingivalis isolates against photokilling action of the photo-activated disinfection.

作者信息

Pourhajibagher Maryam, Ghorbanzadeh Roghayeh, Bahador Abbas

机构信息

Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.

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

出版信息

Lasers Med Sci. 2018 Feb;33(2):337-341. doi: 10.1007/s10103-017-2386-4. Epub 2017 Nov 15.

Abstract

Porphyromonas gingivalis is a significant root canal pathogen capable of causing endodontic infections, which during their treatment may receive sub-lethal doses of photo-activated disinfection (sPAD). As sPAD can influence microbial virulence, this study was designed to evaluate the effect of sPAD on gene expression level of arginine A-specific cysteine proteinase (rgpA), as one of the underlying virulence factors involved in the development of endodontic infection via P. gingivalis strains. To find out the sPAD against 16 clinical isolates of PAD-resistant P. gingivalis that were isolated in vivo, we used toluidine blue O (TBO), methylene blue (MB), and indocyanine green (ICG) as the photosensitizers, which were excited with specific wavelength of light in vitro. Quantitative real-time PCR (qRT-PCR) was then applied to monitor gene expression of rgpA in P. gingivalis isolates to characterize its virulence agent and understand the effect of sPAD on its pathogenicity. Maximal sPAD that could not decrease the count of P. gingivalis isolates were 6.25, 15.6, and 25 μg/mL at fluencies of 171.87, 15.6, and 93.75 J/cm for TBO, ICG, and MB, respectively. ICG-sPAD could suppress the rgpA gene expression about 14-fold, while MB and TBO-mediated sPAD could cause the attenuation of rgpA expression about 4.9- and 11.6-fold, respectively. ICG-sPAD with the maximum ability to reduce rgpA gene expression compared with other photosensitizers can be an appropriate candidate for the treatment of endodontic infections.

摘要

牙龈卟啉单胞菌是一种重要的根管病原体,能够引起牙髓感染,在牙髓感染治疗过程中可能会接受亚致死剂量的光活化消毒(sPAD)。由于sPAD会影响微生物的毒力,本研究旨在评估sPAD对精氨酸A特异性半胱氨酸蛋白酶(rgpA)基因表达水平的影响,rgpA是牙龈卟啉单胞菌菌株引起牙髓感染过程中涉及的潜在毒力因子之一。为了找出针对16株体内分离出的耐PAD牙龈卟啉单胞菌临床分离株的sPAD,我们使用甲苯胺蓝O(TBO)、亚甲蓝(MB)和吲哚菁绿(ICG)作为光敏剂,在体外以特定波长的光激发它们。然后应用定量实时聚合酶链反应(qRT-PCR)监测牙龈卟啉单胞菌分离株中rgpA的基因表达,以表征其毒力因子,并了解sPAD对其致病性的影响。对于TBO、ICG和MB,在光通量分别为171.87、15.6和93.75 J/cm时,不能降低牙龈卟啉单胞菌分离株数量的最大sPAD分别为6.25、15.6和25 μg/mL。ICG-sPAD可使rgpA基因表达抑制约14倍,而MB和TBO介导的sPAD可使rgpA表达分别减弱约4.9倍和11.6倍。与其他光敏剂相比,具有最大降低rgpA基因表达能力的ICG-sPAD可能是治疗牙髓感染的合适候选药物。

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