Selenoprotein W depletion in vitro might indicate that its main function is not as an antioxidative enzyme
Selenoprotein W depletion in vitro might indicate that its main function is not as an antioxidative enzyme
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DOI：
10.1134/s0006297910020100
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发表时间：
2010-03
期刊：
影响因子：
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通讯作者：
Xiao-long Wang;Chuanwei Yang;Kai Xu;Ou-Jv Qin
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作者：
Xiao-long Wang;Chuanwei Yang;Kai Xu;Ou-Jv Qin
在硒蛋白W （SelW）存在或不存在的情况下，检查骨骼肌细胞的抗氧化稳态对于理解SelW在抗氧化系统中的重要性是必要的。通过将合成的小干扰RNA引入小鼠骨骼肌细胞系C2C12 (C3H)，实现了RNA干扰对SelW的消耗。通过实时逆转录- pcr、Western blotting、流式细胞术、荧光染色、细胞活力和谷胱甘肽检测进行转染筛选。SelW表达和mRNA水平分别下调62.1%和72.4%。此外，selw -贫细胞的急性细胞毒性和约36%的凋亡率表明RNA干扰是成功的。谷胱甘肽过氧化物酶、超氧化物歧化酶和过氧化氢酶活性、总抗氧化能力和谷胱甘肽水平分别比未缺硒细胞提高47.6%、103.0、31.0、205.6和30.0% （P< 0.05）。因此，SelW对肌肉细胞的抗氧化系统很重要。然而，由于氧化应激不是导致SelW缺失细胞凋亡的原因，SelW的缺失可以通过其他细胞内抗氧化酶来补偿。因此，SelW在肌肉细胞中的主要功能并不在抗氧化系统中。
Examination of the antioxidative homeostasis in skeletal muscle cells in the presence or absence of selenoprotein W (SelW) is necessary to understand the importance of SelW in the antioxidative system. Depletion of SelW by RNA interference was achieved by introducing a synthetic small interfering RNA into the mouse skeletal muscle cell line C2C12 (C3H). Transfectant screening was performed by real-time reverse transcription-PCR, Western blotting, flow cytometry, fluorescence staining, cell viability, and glutathione assays. SelW expression and mRNA levels were downregulated by 62.1 and 72.4%, respectively. In addition, acute cytotoxicity and an apoptosis rate of ∼36% in SelW-depleted cells demonstrated that RNA interference was successful. As compared with non-SelW-depleted cells, the enzyme activities of glutathione peroxidase, superoxide dismutase, and catalase and total antioxidative capability and glutathione level increased by 47.6, 103.0, 31.0, 205.6, and 30.0%, respectively (P< 0.05). Thus, SelW is important for the antioxidative system of muscle cells. Depletion of SelW, however, could be compensated by other intracellular antioxidative enzymes because oxidative stress was not the causative factor for apoptosis in SelW-depleted cells. Thus, the main function of SelW in muscle cells is not in the antioxidative system.