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

作　　者：陈卉 谢安劼 游毅 陆逸莹 王长德 CHEN Hui;XIE Anjie;YOU Yi;LU Yiying;WANG Changde(Department of Neurology,Shanghai Traditional Chinese Medicine-Integrated Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200082,China)

机构地区：[1]上海中医药大学附属上海市中西医结合医院脑病科,上海200082

出　　处：《上海中医药大学学报》2023年第6期34-40,65,共8页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：上海市进一步加快中医药事业发展三年行动计划中医药诊疗模式创新试点建设项目(ZY[2018-2020]-FWTX-6013);上海市虹口区卫健委中医药科学研究项目(HKQ-ZYY-2021-04,HKQ-ZYY-2020-15)。

摘　　要：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.

关 键 词：红花黄色素A 氧化应激 Nrf2通路 PC12细胞 

分 类 号：R285.5[医药卫生—中药学]