Abstract：
OBJECTIVE: Schwann cells are the main cells for myelination and regeneration of peripheral nerves. Idebenone is a synthetic antioxidant used to treat central nervous system diseases. The aim of the study is to determine whether idebenone can protect Schwann cells and increase cell activity under conditions of oxidative stress caused by hydrogen peroxide (H2O2) in vitro.
METHODS: In this experimental study, Schwann cells were pre-treated with various concentrations of idebenone and H2O2; after determining the appropriate doses, the cells were treated with 10 μM idebenone for 48 hours and 1000 μM H2O2 for the last two hours. The malondialdehyde (MDA) level, and activity of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were assessed by ELISA. Cell viability was assessed by the MTT assay. Western blot analysis was conducted to determine the expressions of myelin protein zero (MPZ) and peripheral myelin protein 22 (PMP22), and expression ratio of the Bax/Bcl-2 proteins. The percentage of cell apoptosis was evaluated by annexin V staining using flow cytometry.
RESULTS: Schwann cells under oxidative stress conditions caused by H2O2 and treated with idebenone had increased cell viability; increased SOD, CAT, and GPx activity; and increased expressions of the MPZ and PMP22 proteins. There was a decreased level of MDA, decreased expression ratio of Bax/Bcl-2 proteins, and a decrease in the percentage of apoptotic cells stained with Annexin V.
CONCLUSIONS: The appropriate dose of idebenone may improve both survival and function of Schwann cells exposed to H2O2 by reducing oxidative stress and apoptosis.
METHODS: In this experimental study, Schwann cells were pre-treated with various concentrations of idebenone and H2O2; after determining the appropriate doses, the cells were treated with 10 μM idebenone for 48 hours and 1000 μM H2O2 for the last two hours. The malondialdehyde (MDA) level, and activity of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were assessed by ELISA. Cell viability was assessed by the MTT assay. Western blot analysis was conducted to determine the expressions of myelin protein zero (MPZ) and peripheral myelin protein 22 (PMP22), and expression ratio of the Bax/Bcl-2 proteins. The percentage of cell apoptosis was evaluated by annexin V staining using flow cytometry.
RESULTS: Schwann cells under oxidative stress conditions caused by H2O2 and treated with idebenone had increased cell viability; increased SOD, CAT, and GPx activity; and increased expressions of the MPZ and PMP22 proteins. There was a decreased level of MDA, decreased expression ratio of Bax/Bcl-2 proteins, and a decrease in the percentage of apoptotic cells stained with Annexin V.
CONCLUSIONS: The appropriate dose of idebenone may improve both survival and function of Schwann cells exposed to H2O2 by reducing oxidative stress and apoptosis.
摘要：
目的：雪旺细胞是周围神经髓鞘形成和再生的主要细胞。艾地苯醌是一种用于治疗中枢神经系统疾病的合成抗氧化剂。该研究的目的是确定艾地苯醌是否可以在体外由过氧化氢（H2O2）引起的氧化应激条件下保护雪旺细胞并增加细胞活性。
方法：在本实验研究中，施旺细胞用不同浓度的艾地苯醌和H2O2预处理;确定合适的剂量后，用10μM艾地苯醌处理细胞48小时,用1000μMH2O2处理最后2小时。丙二醛（MDA）水平，和超氧化物歧化酶（SOD）的活性，过氧化氢酶(CAT),和谷胱甘肽过氧化物酶（GPx）通过ELISA评估。通过MTT测定评估细胞活力。进行Westernblot分析以确定髓鞘蛋白零（MPZ）和外周髓鞘蛋白22（PMP22）的表达。Bax/Bcl-2蛋白的表达比例。使用流式细胞术通过膜联蛋白V染色评估细胞凋亡的百分比。
结果：施万细胞在H2O2引起的氧化应激条件下，用艾地苯醌处理后，细胞活力增加；SOD增加，CAT,和GPx活性;以及MPZ和PMP22蛋白的表达增加。MDA水平下降，Bax/Bcl-2蛋白表达率降低，膜联蛋白V染色的凋亡细胞百分比降低。
结论：适当剂量的艾地苯醌可以通过减少氧化应激和凋亡来改善暴露于H2O2的雪旺细胞的存活和功能。
方法：在本实验研究中，施旺细胞用不同浓度的艾地苯醌和H2O2预处理;确定合适的剂量后，用10μM艾地苯醌处理细胞48小时,用1000μMH2O2处理最后2小时。丙二醛（MDA）水平，和超氧化物歧化酶（SOD）的活性，过氧化氢酶(CAT),和谷胱甘肽过氧化物酶（GPx）通过ELISA评估。通过MTT测定评估细胞活力。进行Westernblot分析以确定髓鞘蛋白零（MPZ）和外周髓鞘蛋白22（PMP22）的表达。Bax/Bcl-2蛋白的表达比例。使用流式细胞术通过膜联蛋白V染色评估细胞凋亡的百分比。
结果：施万细胞在H2O2引起的氧化应激条件下，用艾地苯醌处理后，细胞活力增加；SOD增加，CAT,和GPx活性;以及MPZ和PMP22蛋白的表达增加。MDA水平下降，Bax/Bcl-2蛋白表达率降低，膜联蛋白V染色的凋亡细胞百分比降低。
结论：适当剂量的艾地苯醌可以通过减少氧化应激和凋亡来改善暴露于H2O2的雪旺细胞的存活和功能。
