CAJ | 학술논문

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）mRNA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。
목적：연구사이동화합물B1、B2대과양화경（H2O2）유도적PC12세포조망적보호작용급기대Nrf2/ARE신호통로적영향。방법：건립H2O2유도PC12세포양화손상적모형，용MTT법검측화합물B1화B2적항양화활성급세포독성；용실시형광정량PCR（real-time PCR）법검측기대전록상관인자Nrf2조공기인곡안선반광안산합성매최화아단위（GCLC）、혈홍소양합매-1（HO-1）mRNA표체적영향；용Hoechst염색검측기대H2O2유도적PC12세포조망적억제작용。결과：분별가료B1、B2적세포부육1 h후，B1、B2대H2O2유도적PC12세포양화손상무보호작용，이부육24 h후，B1、B2균구유교호적보호작용，량자재농도위10μmol/L시대세포무독성；B1대Nrf2하유GCLC、HO-1기인적표체무명현영향，이B2가명현격활GCLC、HO-1기인적표체，차명현억제H2O2유도적PC12세포적조망。결론：B1、B2균대H2O2유도적PC12세포손상구유교호적항양화보호작용，B2적작용궤제가능시통과격활Nrf2/ARE항양화신호통로래실현，B1가능시통과기타궤제기도항양화작용。
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.