酰化4-羟基查耳酮对H_2O_2诱导的PC12细胞凋亡的抗氧化保护作用及初步机制  

Antioxidant protection against PC12 cells apoptosis induced by H_2O_2 and the preliminary mechanism of acylated 4-hydroxy chalcones

作　　者：王亚男[1] 沈莱莱 王占坤[3] 周嘉蓓 程婵婵[2] 胡琼琼[1] 方鹏程[1] 仇佩虹[2] 吴建章[2] 梁广[2] 

机构地区：[1]温州医科大学检验医学院生命科学学院,浙江温州325035 [2]温州医科大学药学院化学生物学研究中心,浙江温州325035 [3]温州医科大学体育科学学院,浙江温州325035

出　　处：《温州医学院学报》2015年第1期26-30,共5页Journal of Wenzhou Medical College

基　　金：国家自然科学基金资助项目(81272462);浙江省自然科学基金资助项目(Y13H300005);浙江省大学生科技创新活动计划(新苗人才计划)项目(2013R413032)

摘　　要：目的:研究查尔酮化合物B1、B2对过氧化氢(H2O2)诱导的PC12细胞凋亡的保护作用及其对Nrf2/ARE信号通路的影响。方法:建立H2O2诱导PC12细胞氧化损伤的模型,用MTT法检测化合物B1和B2的抗氧化活性及细胞毒性;用实时荧光定量PCR(real-time PCR)法检测其对转录相关因子Nrf2调控基因谷氨酰半胱氨酸合成酶催化亚单位(GCLC)、血红素氧合酶-1(HO-1)m RNA表达的影响;用Hoechst染色检测其对H2O2诱导的PC12细胞凋亡的抑制作用。结果:分别加了B1、B2的细胞孵育1 h后,B1、B2对H2O2诱导的PC12细胞氧化损伤无保护作用,而孵育24 h后,B1、B2均具有较好的保护作用,两者在浓度为10μmol/L时对细胞无毒性;B1对Nrf2下游GCLC、HO-1基因的表达无明显影响,而B2可明显激活GCLC、HO-1基因的表达,且明显抑制H2O2诱导的PC12细胞的凋亡。结论:B1、B2均对H2O2诱导的PC12细胞损伤具有较好的抗氧化保护作用,B2的作用机制可能是通过激活Nrf2/ARE抗氧化信号通路来实现,B1可能是通过其他机制起到抗氧化作用。Objective： To investigate the protective function of previously synthesized and screened chalcones B1 and B2 against PC12 cells apoptosis induced by H2O2, and their impact on Nrf2/ARE signaling pathway.Methods： A model that PC12 cells were damaged by H2O2 was established, and MTT assay was used to detect the antioxidant activity as well as cytotoxicity of compounds B1 and B2. Real-time PCR was used to determine mRNA expression of Nrf-2 regulated genes GCLC and HO-1. Hoechst staining was applied to test the inhibitory ability against H2O2-mediated PC12 cells apoptosis.Results： B1 and B2 showed no protection on PC12 cells induced by H2O2 in case of 1 hour incubation, while they represented potent antioxidant efifciency af-ter incubation for 24 hours, and both were nontoxic at 10 μmol/L. B1 showed no obvious effect on the expression of GCLC and HO-1 which were Nrf2-regulated downstream genes. B2 signiifcantly prevented the PC12 cells apoptosis caused by H2O2 with activating the expression of GCLC and HO-1.Conclusion： B1 and B2 show a signiifcant antioxidant activity against H2O2-induced PC12 cells injury. The possible mechanism of B2 may be its activation of Nrf2/ARE signaling pathways while B1 may perform the antioxidant action via other route.

关 键 词：查尔酮 合成 抗氧化 H2O2 Nrf2/ARE信号通路 

分 类 号：R966[医药卫生—药理学]