Wang Yuming, Wang Wei, Qiu Enduo
Department of bone and soft-tissue tumor surgery, Cancer Hospital of China Medical University, Liaoning Cancer Hospital & Institute, No.44 Xiaoheyan Road, Dadong District, Shenyang, Liaoning Province, 110042, PR China.
Saudi J Biol Sci. 2017 May;24(4):837-842. doi: 10.1016/j.sjbs.2016.12.004. Epub 2016 Dec 18.
Current study was aimed to investigate the effect of dihydromyricetin on hydrogen peroxide induced oxidative stress in the osteosarcoma cells. MTT assay showed that hydrogen peroxide treatment at a concentration of 100 μM caused a significant ( < 0.005) reduction in the viability of MG63 cells. However, reduction in cell viability caused by 100 μM concentration of hydrogen peroxide was completely prevented on incubation with 30 μM dose of dihydromyricetin. Treatment with 100 μM concentration of hydrogen peroxide for 24 h led to condensation of chromatin material, rounding of cell shape and detachment of cells. The results from flow cytometry using annexin V-FITC and PI double staining showed apoptosis induction in 47.84 ± 5.21% cells on treatment with 100 μM concentration of hydrogen peroxide compared to 2.32 ± 0.54% in controlcells. The apoptotic alterations in MG63 cell morphology were prevented significantly on pre-treatment with 30 μM doses of dihydromyricetin for 48 h. Annexin V-FITC and PI staining showed reduction of hydrogen peroxide induced apoptotic cell percentage to 3.07 ± 0.86% on pre-treatment of MG63 cells with 30 μM dose of dihydromyricetin. Western blot analysis showed a significant increase in the activation of caspase-3 and -9 on treatment of MG63 cells for 24 h with 100 μM concentration of hydrogen peroxide. The expression level of Bcl-2 was decreased significantly by 100 μM concentration of hydrogen peroxide in MG63 cells. However, pre-treatment of MG63 cells with 30 μM dose of dihydromyricetin for 48 h significantly prevented hydrogen peroxide induced increase in caspase-3 and -9 levels and reduction in Bcl-2 level. Thus dihydromyricetin prevents hydrogen peroxide induced reduction in viability and induction of apoptosis in MG63 cells through down-regulation of caspase activation and up-regulation of Bcl-2 levels.
当前研究旨在探讨二氢杨梅素对过氧化氢诱导的骨肉瘤细胞氧化应激的影响。MTT 法检测显示，100 μM 浓度的过氧化氢处理导致 MG63 细胞活力显著降低（<0.005）。然而，在与 30 μM 剂量的二氢杨梅素共同孵育后，100 μM 浓度过氧化氢所导致的细胞活力降低被完全阻止。用 100 μM 浓度的过氧化氢处理 24 小时导致染色质物质凝聚、细胞形状变圆以及细胞脱离。使用膜联蛋白 V-FITC 和碘化丙啶（PI）双重染色的流式细胞术结果显示，与对照细胞中的 2.32±0.54%相比，用 100 μM 浓度的过氧化氢处理后，47.84±5.21%的细胞发生凋亡诱导。在用 30 μM 剂量的二氢杨梅素预处理 48 小时后，MG63 细胞形态的凋亡改变得到显著预防。膜联蛋白 V-FITC 和 PI 染色显示，在用 30 μM 剂量的二氢杨梅素预处理 MG63 细胞后，过氧化氢诱导的凋亡细胞百分比降至 3.07±0.86%。蛋白质免疫印迹分析显示，用 100 μM 浓度的过氧化氢处理 MG63 细胞 24 小时后，半胱天冬酶 -3 和 -9 的激活显著增加。在 MG63 细胞中，100 μM 浓度的过氧化氢显著降低了 Bcl-2 的表达水平。然而，用 30 μM 剂量的二氢杨梅素预处理 MG63 细胞 48 小时可显著预防过氧化氢诱导的半胱天冬酶 -3 和 -9 水平升高以及 Bcl-2 水平降低。因此，二氢杨梅素通过下调半胱天冬酶激活和上调 Bcl-2 水平，预防过氧化氢诱导的 MG63 细胞活力降低和凋亡诱导。
