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具有谷胱甘肽过氧化物酶活性的人含硒单链可变片段保护NIH3T3成纤维细胞免受氧化损伤

[Human selenium-containing single-chain variable fragment with glutathione peroxidase activity protects NIH3T3 fibroblast against oxidative damage].

作者信息

Huo R, Yang L, Zhang T G, Wei J Y

机构信息

College of Pharmaceutical Science, Jilin University, Changchun, 130021 P. R. China.

China-Japan Union Hospital, Jilin University, Changchun, 130033 P. R. China.

出版信息

Mol Biol (Mosk). 2017 May-Jun;51(3):483-489. doi: 10.7868/S0026898417030077.

Abstract

Ultraviolet B (UVB medium wave, 280-315 nm) induces cellular oxidative damage and apoptosis by producing reactive oxygen species (ROS). Glutathione peroxidase functions as an antioxidant by catalyzing the reduction of hydrogen peroxide, the more important member of reactive oxygen species. A human selenium-containing single-chain variable fragment (se-scFv-B3) with glutathione peroxidase activity of 1288 U/μmol was generated and investigated for its antioxidant effects in UVB-induced oxidative damage model. In particular, cell viability, lipid peroxidation extent, cell apoptosis, the change of mitochondrial membrane potential, caspase-3 activity and the levels of intracellular reactive oxygen species were assayed. Human se-scFv-B3 protects NIH3T3 cells against ultraviolet B-induced oxidative damage and subsequent apoptosis by prevention of lipid peroxidation, inhibition of the collapse of mitochondrial membrane potential as well as the suppression of the caspase-3 activity and the level of intracellular ROS. It seems that antioxidant effects of human se-scFv-B3 are mainly associated with its capability to scavenge reactive oxygen species, which is similar to that of the natural glutathione peroxidase.

摘要

紫外线B(UVB中波,280 - 315纳米)通过产生活性氧(ROS)诱导细胞氧化损伤和凋亡。谷胱甘肽过氧化物酶通过催化过氧化氢(活性氧中更重要的成员)的还原发挥抗氧化剂的作用。生成了一种具有1288 U/μmol谷胱甘肽过氧化物酶活性的人含硒单链可变片段(se-scFv-B3),并在UVB诱导的氧化损伤模型中研究其抗氧化作用。特别地,检测了细胞活力、脂质过氧化程度、细胞凋亡、线粒体膜电位变化、半胱天冬酶-3活性以及细胞内活性氧水平。人se-scFv-B3通过预防脂质过氧化、抑制线粒体膜电位崩溃以及抑制半胱天冬酶-3活性和细胞内ROS水平,保护NIH3T3细胞免受紫外线B诱导的氧化损伤和随后的凋亡。人se-scFv-B3的抗氧化作用似乎主要与其清除活性氧的能力有关,这与天然谷胱甘肽过氧化物酶相似。

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