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蓝藻腺苷同型半胱氨酸酶的抗氧化分子机制及其在成纤维细胞中的创伤愈合过程。

Antioxidant molecular mechanism of adenosyl homocysteinase from cyanobacteria and its wound healing process in fibroblast cells.

机构信息

SRM Research Institute, SRM Institute of Science and Technology, Kattankulathur, Chennai, Tamil Nadu, 603 203, India.

Lab PCN 206, Microbiology Division, CSIR-Central Drug Research Institute, B.S. 10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow, Uttar Pradesh, 226 031, India.

出版信息

Mol Biol Rep. 2020 Mar;47(3):1821-1834. doi: 10.1007/s11033-020-05276-y. Epub 2020 Jan 27.

Abstract

An antioxidant molecule namely, adenosyl homocysteinase (AHc) was identified from the earlier constructed transcriptome database of Spirulina, where it was cultured in a sulphur deprived condition. From the AHc protein, a small peptide NL13 was identified using bioinformatics tools and was predicted to have antioxidant property. Further, the peptide was synthesised and its antioxidant mechanism was addressed at molecular level. NL13 was subjected to various antioxidant assays including DPPH assay, HARS assay, SARS Assay, NO assay and ABTS assay, where NL13 exhibited significant (P < 0.05) potential antioxidant activity compared to its antioxidant control, Trolox. Cytotoxicity was performed on Human whole blood and the cell viability was performed on VERO fibroblast cells. In both assays, it was found that NL13 did not exhibit any cytotoxic effect towards the cells. Further, the intracellular ROS was performed on Multimode reader followed by imaging on fluorescence microscope which showed scavenging activity even at lower concentration of NL13 (31.2 µM). An effective wound healing property of NL13 on VERO cells was confirmed by analysing the cell migration rate at two different time intervals (24 and 48 h). Overall, the study shows that NL13 peptide scavenges the intracellular oxidative stress.

摘要

从先前构建的螺旋藻转录组数据库中鉴定出一种抗氧化分子,即腺苷同型半胱氨酸酶(AHc),在该数据库中,螺旋藻在缺乏硫的条件下进行培养。从 AHc 蛋白中,使用生物信息学工具鉴定出一种小肽 NL13,并预测其具有抗氧化特性。此外,还合成了该肽,并在分子水平上研究了其抗氧化机制。NL13 进行了各种抗氧化测定,包括 DPPH 测定、HARS 测定、SARS 测定、NO 测定和 ABTS 测定,结果表明 NL13 与抗氧化对照 Trolox 相比具有显著的(P<0.05)潜在抗氧化活性。在人全血和 VERO 成纤维细胞上进行了细胞毒性测定。在这两种测定中,都发现 NL13 对细胞没有任何细胞毒性作用。此外,在多模式阅读器上进行细胞内 ROS 测定,然后在荧光显微镜下进行成像,结果表明 NL13 即使在较低浓度(31.2µM)下也具有清除活性。通过分析在两个不同时间间隔(24 和 48 小时)的细胞迁移率,证实了 NL13 在 VERO 细胞上具有有效的伤口愈合特性。总的来说,该研究表明 NL13 肽可清除细胞内的氧化应激。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d82b/7223595/ffe865d62391/11033_2020_5276_Fig1_HTML.jpg

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