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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100μmol·L-1)预孵育12 h,加入H2O2(150μmol·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100μmol·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。
To observe the effect of baicalin (BL) on hydrogen peroxide-induced SH-SY5Y cell injury and to investigate whether SIRT1 is the target of its action and its possible mechanism. The cultured human neuroblastoma cell line SH-SY5Y cells were pre-incubated with baicalin (25, 50 and 100 μmol·L-1) for 12 h, then induced by H2O2 (150 μmol·L-1) for 24 h, Cell viability was measured by MTT colorimetric assay. LDH leakage rate and nitric oxide (NO) content were determined in the culture medium. Apoptosis was detected by flow cytometry and Caspase -3 activity, RT-PCR detection of SIRT1 levels. The results showed that compared with the H2O2 model group, the cell viability of baicalin (50 and 100μmol·L-1) group increased (P <0.05, P <0.01), while the release of LDH, NO and the number of apoptotic cells <0.05, P <0.01), and the expression of Caspase-3 decreased; the expression of SIRT1 mRNA was significantly increased (P <0.05, P <0.01). Baicalin has a protective effect on SY5Y cells injured by H2O2, which can reduce the damage of nerve cells and inhibit the apoptosis of cells. Its mechanism may be through up-regulating the expression of SIRT1 and inhibiting the expression of Caspase-3.