灯盏花素对缺氧/复氧诱导的皮质神经元凋亡的影响

Effects of scutellarin on apoptosis of cortical neurons induced by hypoxia/reoxygenation

目的探讨灯盏花素对缺氧/复氧损伤大鼠皮质神经元的保护作用,并探讨其机制。方法培养大鼠皮质神经元,造成缺氧/复氧损伤模型,分为对照组、模型组、灯盏花素低浓度组及灯盏花素高浓度组,CCK-8法观察细胞活性,检测各组细胞上清液乳酸脱氢酶(LDH)及细胞超氧化物歧化酶(SOD)、丙二醛(MDA)含量变化,流式细胞技术检测细胞内活性氧(ROS)水平,Hoechst染色检测凋亡细胞比例,Western blot分析Bcl-2、Bax的变化。结果 CCK-8结果显示,用1、10μmol/L浓度的灯盏花素处理后,皮质神经元存活率分别提高到59.87%±4.53%和69.93%±5.27%,均高于模型组(36.68%±4.79%)(P<0.05);2′,7′-二氯二氢荧光素二乙酸酯(H2DCFDA)荧光探针染色结果显示,灯盏花素抑制了神经元细胞内ROS水平及细胞凋亡,增强了Bcl-2蛋白的表达,降低了Bax蛋白的表达。结论灯盏花素可以抑制缺氧/复氧诱导的神经元氧化损伤及凋亡的发生,为预防和治疗缺血性脑卒中及退行性神经疾病提供了理论支持。

Objective To investigate the protective effect of scutellarin on cortical neurons damaged by hypoxia/reoxygenation and discuss its mechanism.Methods Cortical neurons were cultured and hypoxia/reoxygenation models were constructed,and the cells were divided into control group,model group,low-concentration scutellarin group and high-concentration scutellarin group.The cell survival rate was detected by CCK-8 colorimetry;the changes of lactate dehydrogenase(LDH) in cell supernatant and the levels superoxide dismutase(SOD)and malondialdehyde(MDA)in cells were measured;the level of intracellular reactive oxygen species(ROS) was detected by flow cytometry technique;the apoptotic cells proportion was detected by Hoechst staining;the changes in Bcl-2 and Bax were analyzed by Western blot.Results The CCK-8 results showed that cortical neurons survival rate increased to 59.87%±4.53%and 69.93%±5.27% after being treated with 1 μmol/L and 10 μmol/L scutellarin,which was higher than that of model group(36.68%±4.79%,P<0.05);the H2DCFDA fluorescent probe dying showed that scutellarin reduced ROS generation in neurons and cell apoptosis,enhanced the Bcl-2 protein expression,and reduced the expression of Bax.Conclusion Scutellarin can inhibit hypoxia/reoxygenation-induced oxidative damage and apoptosis in cortical neurons,which provides theoretical support for the prevention and treatment of ischemic stroke and degenerative neurological disease.