基于Nrf2通路探讨红花黄色素A对H_(2)O_(2)诱导PC12细胞株氧化应激损伤的保护作用

Protective effect study of safflor yellow A against H_(2)O_(2)⁃induced oxidative stress injury in pheochromocytoma PC12 Cells

目的:探讨红花黄色素A(SYA)对氧化应激损伤的保护作用,并探讨其可能的分子机制。方法:采用过氧化氢(H_(2)O_(2))和SYA处理嗜铬细胞瘤细胞株PC12;采用CCK-8实验测定细胞增殖,采用CM-H2DCF-DA染色细胞内活性氧(ROS);采用Western blot检测细胞凋亡相关蛋白B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、胱天蛋白酶-3(Caspase-3)、切割型胱天蛋白酶-3(cleaved Caspase-3)和氧化应激调节因子核因子E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)的水平。结果:CCK-8检测结果显示,经SYA处理后,H_(2)O_(2)诱导的氧化应激PC12细胞的存活率明显提高。荧光显微镜(CM-H2DCF-DA染色)显示SYA可以通过减少细胞内ROS来降低H_(2)O_(2)诱导的PC12细胞凋亡。Western blot检测显示,SYA可以上调经H_(2)O_(2)处理的PC12细胞中Nrf2和Bcl-2的表达,同时可以下调Bax、Caspase-3和cleaved Caspase-3的表达。结论:SYA可保护神经细胞免受氧化应激损伤,激活Nrf2通路可能是其相关作用机制。

Objective:To explore the protective effect of safflor yellow A(SYA)on oxidative stress injury and its possible molecular mechanism.Methods:Pheochromocytoma PC12 cells were treated with H_(2)O_(2)and SYA.Cell proliferation was determined by Cell counting kit-8(CCK-8)assay.Reactive oxygen species(ROS)was stained with 5-(and 6)-Chloromethyl-2′,7′-Dichloro-dihydrofluorescein diacetate(CM-H2DCF-DA).The levels of apoptosis-related proteins of Bax,Bcl-2,Caspase-3,cleaved Caspase-3 and oxidative stress regulators Nrf2,HO-1 were detected by Western blot.Results:CCK-8 assay results showed the survival rate of H_(2)O_(2)-induced oxidative stress PC12 cells was significantly increased after SYA treatment.Fluorescence microscope(CM-H2DCF-DA staining)indicated the H_(2)O_(2)-induced apoptosis in PC12 cells was decreased by reducing intracellular ROS because of SYA.Western blot assay showed the expressions of Nrf2 and Bcl-2 were up-regulated by SYA while the expressions of Bax,Caspase-3,cleaved Caspase-3 were down-regulated in H_(2)O_(2)-induced PC12 cells.Conclusions:SYA can protect nerve cells from oxidative stress injury,and its mechanism may be related to the activation of the Nrf2 pathway.