现代中西医结合杂志2025，Vol.34Issue(16)：2228-2236,9.DOI:10.3969/j.issn.1008-8849.2025.16.006
银杏二萜内酯调节Nrf2/HO-1信号通路对皮质神经元氧糖剥夺/复氧复糖损伤的影响
Effect of Ginkgo biloba diterpenoid lactones on cortical neuron with oxygen glucose deprivation/reoxygenation injury via regulating Nrf2/HO-1 signaling pathway
摘要
Abstract
Objective It is to investigate the effects and potential mechanisms of Ginkgo biloba diterpenoid lactones(GDL)on cortical neuron with oxygen glucose deprivation/reoxygenation(OGD/R)injury based on nuclear factor E2 relat-ed factor 2(Nrf2)/heme oxygenase 1(HO-1)signal pathway.Methods The cortical neurons of newborn 24 hour neonatal SD rats were isolated in a sterile environment.After 7 days of culture,the cortical neurons with a purity>90%and in the logarithmic growth phase were selected and divided into normal group,model group,GDL group,GDL+Nrf2 agonist group and GDL+Nrf2 inhibitor group.The cortical neurons of the normal group were normally cultured,and the neurons of the other groups were prepared with hypoxia(5%CO2+95%N2)for4 hours and reoxygenation(5%CO2+95%air)for24 hours after cultured for 24 h to establish models of cortical neuron OGD/R injury.The cortical neurons of the control group and model group were cultured in replaced conventional medium containing no drugs,the neurons of the GDL group,GDL+Nrf2 agonist group and GDL+Nrf2 inhibitor group were cultured in replaced medium containing 50 μg/mL GDL,50 μg/mL GDL+1.6 μg/mL TBHQ,50 μg/mL GDL+5 μg/mL ML385,respectively.After culture for 24 hours,the activity corti-cal neurons was determined by CCK-8,and the apoptosis rate of neurons was determined by flow cytometry,the content of reactive oxygen species(ROS)in cortical neurons was measured by dichlorodihy drofluorescein acetoacetate(DCFH-DA)fluorescent probe method,the contents of oxidative stress indicators[content of malondialdehyde(MDA),activities of su-peroxide dismutase(SOD)and glutathione peroxidase(GSH-Px)]and the levels of inflammatory factors[tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),monocyte chemotactic protein-1(MCP-1)]in the culture medium were measured by ELISA,the mRNA expressions of Nrf2,HO-1 in cortical neurons were detected by RT-PCR,and the protein expressions of Nrf2,HO-1,nuclear factor-κB p65(NF-κB p65),p-NF-κB p65,Bcl-2,Bax,Caspase-3,Cleaved Caspase-3 were detected by Western blot.Results Compared with the normal group,the activity of cortical neuron,activi-ties of SOD and GSH-Px,relative mRNA expressions of Nrf2 and HO-1,and relative protein expressions of Nrf2,HO-1,Bcl-2 in the model group were significantly decreased(all P<0.05),while the apoptosis rate of cortical neuron,contents of ROS,MDA,TNF-α,IL-1β,MCP-1 and the relative protein expressions of p-NF-κB p65,Bax,Cleaved Caspase-3,and p-NF-κB p65/NF-κB p65,Bax/Bcl-2,Cleaved Caspase-3/Caspase-3 were significantly increased(all P<0.05).Com-pared with the model group,the activity of cortical neuron,activities of SOD and GSH-Px,relative mRNA expressions of Nrf2 and HO-1,and relative protein expressions of Nrf2,HO-1,Bcl-2 in the GDL group and GDL+Nrf2 agonist group were significantly increased(all P<0.05),while the apoptosis rate of cortical neuron,contents of ROS,MDA,TNF-α,IL-1β,MCP-1 and the relative protein expressions of p-NF-κB p65,Bax,Cleaved Caspase-3,and p-NF-κB p65/NF-κB p65,Bax/Bcl-2,Cleaved Caspase-3/Caspase-3 were significantly decreased(all P<0.05).Compared with the GDL group,the effects on the above indicators were more significant in the GDL+Nrf2 agonist group(all P<0.05),converse-ly,the effects were significantly attenuated in the GDL+Nrf2 inhibitor group(all P<0.05).Conclusion GDL can inhibit OGD/R injury in cortical neurons,the mechanism may be related to activating Nrf2/HO-1 signaling pathway to inhibit oxi-dative stress,inflammation and apoptosis.关键词
皮质神经元/银杏二萜内酯/氧糖剥夺/复氧复糖/核因子E2相关因2/血红素加氧酶1/氧化应激/炎症Key words
cortical neurons/Ginkgo biloba diterpenoid lactone/oxygen glucose deprivation/reoxygenation/Nrf2/HO-1/oxidative stress/inflammation分类
医药卫生引用本文复制引用
李晓燕,霍好利..银杏二萜内酯调节Nrf2/HO-1信号通路对皮质神经元氧糖剥夺/复氧复糖损伤的影响[J].现代中西医结合杂志,2025,34(16):2228-2236,9.基金项目
河北省2022年度医学科学研究课题计划项目(20220570) （20220570）
