黄芩苷通过上调SIRT1保护SH-SY_5Y氧化应激的损伤

Effects of baicalin against oxidative stress injury of SH-SY_5Y cells by up-regulating SIRT1

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。

This study is to investigate the effect of baicalin（BL） against oxidative injury stress of SH-SY 5 Y cells induced by H 2 O 2 and the possible mechanism.SH-SY 5 Y cells were pre-incubated with baicalin（25,50,and 100 μmol·L 1） for 12 h prior to exposure to H 2 O 2 （150 μmol·L 1） for 24 h.The viability of SH-SY 5 Y 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-SY 5 Y cells.This study results suggest that baicalin could inhibit the H 2 O 2-induced neuronal apoptosis.The further mechanism studies show that baicalin inhibit apoptosis via reducing Caspase-3 expression and up-regulating SIRT1.