重组人金属硫蛋白-Ⅲα短肽对UVB致HaCaT细胞氧化损伤的缓解作用
Allievation Effects of Recombinant Human Metallothionein Ⅲα Peptide on Oxidative Damage of HaCaT Cells Induced by UVB
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摘要: 利用体外自由基清除实验研究重组人金属硫蛋白Ⅲα短肽（rh-MT-Ⅲα ）自由基清除能力，建立紫外线B（UVB）损伤HaCaT细胞模型，使用不同浓度rh-MT-Ⅲα （25～200 μg·mL-1）处理损伤的HaCaT细胞24 h后观察细胞存活率、细胞形态、细胞凋亡及细胞内活性氧（ROS）水平，从细胞水平上评价rh-MT-Ⅲα对UVB致HaCaT细胞氧化损伤的作用。体外自由基清除实验结果显示，随着rh-MT-Ⅲα浓度升高，其对1，1-二苯基-2-苦苯肼（DPPH）自由基清除率也随之增加，清除率为50%时所需rh-MT-Ⅲα浓度（IC50）为184.32 μg·mL-1。抗光氧化损伤实验结果显示，rh-MT-Ⅲα可有效提高UVB损伤后HaCaT细胞的存活率、改善细胞形态并显著降低细胞凋亡及细胞内ROS含量，且在100 μg·mL-1时修复作用最为明显。实验结果表明rh-MT-Ⅲα具有较强自由基清除能力，可有效缓解UVB诱导的HaCaT细胞氧化损伤，且在100 μg·mL-1剂量下作用显著。
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关键词:
- 金属硫蛋白 /
- 紫外线B /
- 人永生化皮肤角质细胞 /
- 抗氧化损伤 /
- 自由基清除
Abstract: The ability of recombinant human metallothionein Ⅲα peptide (rh-MT-Ⅲα) to scavenge free radicals was determined by free radical scavenging experiments in vitro. After the HaCaT cells were damaged by UVB, they were treated with rh-MT-Ⅲα at different doses (25, 50, 100, 200 μg·mL-1) for 24 h. The anti-oxidative function of rh-MT-Ⅲα on oxidative damage of HaCaT cells induced by UVB was evaluated by the alternations of cell viability, cell morphology, cell apoptosis and the production of reactive oxygen species (ROS) in HaCaT cells. The results of free radical scavenging experiment showed that the scavenging rate of 1,1-diphenyl-2-picrylhydrazyl (DPPH) increased as the dose of rh-MT-Ⅲα increased, and the required dose of rh-MT-Ⅲα was 184.32 μg·mL-1 when the scavenging rate was 50%. The results of anti-oxidative function experiment showed that rh-MT-Ⅲα treatment, especially at 100 μg·mL-1, resulted in improved cell viability, more tightly cell connection, and reductions in apoptosis rate and ROS content in HaCaT cells when compared with the positive model group. The findings indicated that the rh-MT-Ⅲα showed strong ability of scavenging free radical, which effectively alleviate UVB-induced oxidative damage in HaCaT cells with the mostly effective dose of 100 μg·mL-1.-
Key words:
- metallothionein /
- UVB /
- HaCaT cells /
- anti-oxidation /
- scavenging free radical
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