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重组人金属硫蛋白-III 可减轻秀丽隐杆线虫中铜和镉诱导的氧化损伤。

Recombinant human metallothionein-III alleviates oxidative damage induced by copper and cadmium in Caenorhabditis elegans.

机构信息

Key Laboratory of Environmental Medicine and Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing, China.

Suzhou Hvha Medical Technology Development Co., Ltd., Changshu, China.

出版信息

J Appl Toxicol. 2023 Aug;43(8):1242-1252. doi: 10.1002/jat.4460. Epub 2023 Mar 14.

Abstract

Recombinant human metallothionein III (rh-MT-III) is a genetically engineered product produced by Escherichia coli fermentation technology. Its molecules contain abundant reducing sulfhydryl groups, which possess the ability to bind heavy metal ions. The present study was to evaluate the binding effects of rh-MT-III against copper and cadmium in vitro and to investigate the antioxidant activity of rh-MT-III using Caenorhabditis elegans in vivo. For in vitro experiments, the binding rates of copper and cadmium were 91.4% and 97.3% for rh-MT-III at a dosage of 200 μg/mL at 10 h, respectively. For in vivo assays, the oxidative stress induced by copper (CuSO , 10 μg/mL) and cadmium (CdCl , 10 μg/mL) was significantly reduced after 72 h of exposure to different doses of rh-MT-III (5-500 μg/mL), indicated by restoring locomotion behavior and growth, and reducing malondialdehyde and reactive oxygen species levels in C. elegans. Moreover, rh-MT-III decreased the deposition of lipofuscin and fat content, which could delay the progression of aging. In addition, rh-MT-III (500 μg/mL) promoted the up-regulation of Mtl-1 and Mtl-2 gene expression in C. elegans, which could enhance the resistance to oxidative stress by increasing the enzymatic activity of antioxidant defense system and scavenging free radicals. The results indicated that supplemental rh-MT-III could effectively protect C. elegans from heavy metal stress, providing an experimental basis for the future application and development of rh-MT-III.

摘要

重组人金属硫蛋白 III(rh-MT-III)是一种通过大肠杆菌发酵技术生产的基因工程产品。其分子中含有丰富的还原巯基,具有结合重金属离子的能力。本研究旨在评估 rh-MT-III 体外结合铜和镉的效果,并通过体内秀丽隐杆线虫研究 rh-MT-III 的抗氧化活性。在体外实验中,rh-MT-III 在 200μg/mL 剂量下孵育 10 小时时,铜和镉的结合率分别为 91.4%和 97.3%。在体内实验中,暴露于不同剂量 rh-MT-III(5-500μg/mL)72 小时后,铜(CuSO4,10μg/mL)和镉(CdCl2,10μg/mL)诱导的氧化应激明显减轻,表现为运动行为和生长恢复,以及秀丽隐杆线虫中丙二醛和活性氧水平降低。此外,rh-MT-III 减少了脂褐素和脂肪含量的沉积,从而延缓衰老的进展。此外,rh-MT-III(500μg/mL)可促进秀丽隐杆线虫 Mtl-1 和 Mtl-2 基因表达上调,通过增加抗氧化防御系统的酶活性和清除自由基来增强对氧化应激的抵抗力。结果表明,补充 rh-MT-III 可有效保护秀丽隐杆线虫免受重金属应激,为 rh-MT-III 的未来应用和开发提供了实验依据。

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