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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 effect of chalcone compounds B1 and B2 on apoptosis of PC12 cells induced by hydrogen peroxide (H2O2) and its effect on Nrf2 / ARE signaling pathway. Methods: The model of H2O2-induced oxidative damage in PC12 cells was established. The anti-oxidative activity and cytotoxicity of compounds B1 and B2 were detected by MTT assay. The effects of transcription factor Nrf2 on the oxidative stress and cytotoxicity of PC12 cells were detected by real-time PCR. (CLC) and heme oxygenase-1 (HO-1) m RNA expression were detected by enzyme-linked immunosorbent assay (ELISA). Hoechst staining was used to detect the inhibitory effect on H2O2-induced PC12 cell apoptosis. Results: After B1 and B2 cells were incubated for 1 h, B1 and B2 had no protective effect on H2O2-induced oxidative damage of PC12 cells, while B1 and B2 had a good protective effect after incubating for 24 h The concentration of 10μmol / L had no toxic effect on the cells; B1 had no effect on the expression of GCLC and HO-1 downstream of Nrf2, while B2 could obviously activate the expression of GCLC and HO-1, and significantly inhibited the H2O2-induced PC12 cells Apoptosis. CONCLUSION: Both B1 and B2 have good antioxidative protective effects on H2O2-induced injury of PC12 cells. The mechanism of B2 may be through activation of Nrf2 / ARE antioxidant signaling pathway. B1 may play an antioxidant role through other mechanisms effect.