黄芩苷通过上调SIRT1保护SH-SY5Y氧化应激的损伤
Effects of baicalin against oxidative stress injury of SH-SY5Y cells by up-regulating SIRT1
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摘要:
观察黄芩苷 (baicalin, BL) 对过氧化氢诱导SH-SY5Y细胞损伤的作用, 并初步探讨SIRT1是否是其发挥作用的靶点及其可能机制。培养的人神经母细胞瘤细胞系SH-SY5Y细胞加入黄芩苷 (25、50及100 μmol·L−1) 预孵育12 h, 加入H2O2 (150 μmol·L−1) 作用24 h诱导产生氧化损伤, 应用MTT比色法检测细胞存活率, 测定培养液中乳酸脱氢酶 (LDH) 漏出率、一氧化氮 (NO) 含量; 并采用流式细胞仪检测细胞凋亡及应用免疫荧光组化法检测Caspase-3的活性、RT-PCR检测SIRT1水平。 结果表明, 与H2O2模型组比较, 黄芩苷 (50和100 μmol·L−1) 组的细胞存活率增高 (P < 0.05、P < 0.01), 而LDH、NO的释放量及细胞凋亡数 (P < 0.05、P < 0.01)、Caspase-3的表达量均减少; SIRT1 mRNA表达明显 (P < 0.05、P < 0.01)。黄芩苷对H2O2损伤的SY5Y细胞具有保护作用, 可减轻神经细胞的损伤、抑制细胞的凋亡, 其作用机制可能是通过上调SIRT1水平而抑制Caspase-3表达实现的。
Abstract:This study is to investigate the effect of baicalin (BL) against oxidative injury stress of SH-SY5Y cells induced by H2O2 and the possible mechanism. SH-SY5Y cells were pre-incubated with baicalin (25, 50, and 100 μmol·L−1) for 12 h prior to exposure to H2O2 (150 μmol·L−1) for 24 h. The viability of SH-SY5Y cells was measured by MTT assay. The contents of LDH and NO were determined. The percentage of apoptotic cells was assessed by flow cytometry (FCM). The content of Caspase-3 was tested by immunofluorescence histochemical method. BL at 50 and 100 μmol·L−1 separately increased the cell viability and up-regulated SIRT1, reduced the contents of LDH, NO, Caspase-3 and the apoptotic percentage of SH-SY5Y cells. This study results suggest that baicalin could inhibit the H2O2-induced neuronal apoptosis. The further mechanism studies show that baicalin inhibit apoptosis via reducing Caspase-3 expression and up-regulating SIRT1.
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