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活性氧和一氧化氮在 HaCaT 细胞抑制生长中的作用。

The Role of Reactive Oxygen Species and Nitric Oxide in the Inhibition of Growth by HaCaT Cells.

机构信息

Department of Dermatology, Huzhou Central Hospital, 198 Hongqi Rd, Huzhou, 313000 Zhejiang, China.

Department of Dermatology, Shunde Hospital, Southern Medical University (The First People's Hospital of Shunde Foshan), No. 1 Jiazi Road, Lunjiao, Shunde District, Foshan City, Guangdong Province, China 528308.

出版信息

Oxid Med Cell Longev. 2020 Feb 12;2020:8548619. doi: 10.1155/2020/8548619. eCollection 2020.

Abstract

() is one of the most important agents of dermatophyte infection in humans. The aim of this experiment was to evaluate the effect of HaCaT cells on , investigate the responsible mechanism of action, and explore the role of reactive oxygen species (ROS) and nitric oxide (NO) in the inhibition of growth by HaCaT cells. The viability of fungi treated with HaCaT cells alone and with HaCaT cells combined with pretreatment with the NADPH oxidase inhibitor (DPI) or the nitric oxide synthase (NOS) inhibitor L-NMMA was determined by enumerating the colony-forming units. NOS, ROS, and NO levels were quantified using fluorescent probes. The levels of the NOS inhibitor asymmetric dimethylarginine (ADMA) were determined by enzyme-linked immunosorbent assay (ELISA). Micromorphology was observed using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). In addition, fungal keratinase activity was assessed by measuring dye release from keratin azure. In vitro fungal viability, keratinase activity, and ADMA content decreased after HaCaT cell intervention, whereas the levels of ROS, NO, and NOS increased. The micromorphology was abnormal. Fungi pretreated with DPI and L-NMMA exhibited opposite effects. HaCaT cells inhibited the growth and pathogenicity of in vitro. A suggested mechanism is that ROS and NO play an important role in the inhibition of growth by HaCaT cells.

摘要

()是人类皮肤癣菌感染最重要的病原体之一。本实验旨在评价 HaCaT 细胞对 的作用,探讨其作用机制,并探讨活性氧(ROS)和一氧化氮(NO)在 HaCaT 细胞抑制 生长中的作用。单独用 HaCaT 细胞和用 HaCaT 细胞与 NADPH 氧化酶抑制剂(DPI)或一氧化氮合酶(NOS)抑制剂 L-NMMA 预处理后处理真菌的真菌活力通过计数形成单位来确定。使用荧光探针定量 NOS、ROS 和 NO 水平。通过酶联免疫吸附试验(ELISA)测定 NOS 抑制剂不对称二甲基精氨酸(ADMA)的水平。使用扫描电子显微镜(SEM)和透射电子显微镜(TEM)观察微观形态。此外,通过测量角蛋白天蓝从角蛋白释放来评估真菌角质酶活性。体外真菌活力、角质酶活性和 ADMA 含量在 HaCaT 细胞干预后降低,而 ROS、NO 和 NOS 水平升高。形态异常。用 DPI 和 L-NMMA 预处理的真菌表现出相反的效果。HaCaT 细胞抑制了体外 的生长和致病性。推测的机制是 ROS 和 NO 在 HaCaT 细胞抑制 生长中起重要作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc01/7038170/da612010d969/OMCL2020-8548619.001.jpg

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