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目的:研究花旗松素对β-淀粉样肽(β-amyloid peptide,Aβ)损伤神经元的保护作用及其机制。方法:从新生乳鼠大脑皮层中分离纯化神经元,与Aβ(5μmol·L-1)、不同浓度花旗松素(20~100μmol·L-1)共同孵育24 h,CCK8法检测细胞存活率,Hoechst 33258染色和Annexin V-FITC/PI检测细胞凋亡,分光光度法检测半胱天冬酶-3(caspase-3)活性的变化。结果:在细胞活力实验中,与正常组相比,模型组细胞活力显著下降(49.2±1.3)%,不同浓度的花旗松素可以提高神经元活力,其中80μmol·L-1花旗松素给药组细胞活力增至(68.7±3.2)%,与模型组相比有极显著差异;细胞凋亡结果显示,与正常组相比较,模型组细胞凋亡率为(50.8±1.5)%(P<0.01),不同浓度的花旗松素给药组可以降低细胞凋亡率,其中40μmol·L-1花旗松素凋亡率为(41.5±2.7)%,与模型组比较呈显著性差异(P<0.05);凋亡酶caspase-3的活性与正常组(0.12±0.02)U·μg-1比较,模型组的caspase-3的活性升高(2.37±0.16)U·μg-1,P<0.01),与模型组相比,40μmol·L-1花旗松素组caspase-3显著降低(1.77±0.07)U·μg-1,P<0.01)。花旗松素能明显提高Aβ损伤神经元的细胞活力,抑制神经元的凋亡,降低凋亡酶caspase-3的活性。结论:花旗松素对Aβ损伤神经元具有保护作用,其机制可能与抑制caspase-3活性从而实现抗凋亡作用有关。
Objective: To study the protective effect and the mechanism of Taxifolin on neurons injured by β-amyloid peptide (Aβ). Methods: Neurons were isolated and purified from neonate neonatal rat cerebral cortex, incubated with Aβ (5μmol·L-1) and different concentrations of Taxol (20-100μmol·L-1) for 24 h. Cell viability was determined by CCK8 assay. Hoechst 33258 staining and Annexin V-FITC / PI to detect apoptosis. The change of caspase-3 activity was detected by spectrophotometry. RESULTS: Compared with the normal group, the viability of model group decreased significantly (49.2 ± 1.3)% in the cell viability test. Different concentrations of Taxifolin could increase the neuron viability. Among them, 80 μmol·L-1 of Taxifolin Compared with the normal group, the apoptotic rate of the model group was (50.8 ± 1.5)% (P <0.01), and the cell viability increased to (68.7 ± 3.2)%, which was significantly different from the model group 0.01). The apoptotic rates of Taxol treated with different concentrations of Taxifolin were lower than that of the model group (41.5 ± 2.7%), which was significantly different from the model group (P < 0.05). Compared with the normal group (0.12 ± 0.02) U · μg-1, the activity of caspase-3 in the model group increased (2.37 ± 0.16) U · μg-1, P <0.01 ), Compared with the model group, caspase-3 in 40μmol·L-1 taxanesignificantly decreased (1.77 ± 0.07) U · μg-1, P <0.01). Taxifolin significantly increased cell viability of Aβ injured neurons, inhibited neuronal apoptosis and decreased the activity of caspase-3, a apoptotic enzyme. CONCLUSION: Taxifolin plays a protective role in Aβ-injured neurons, which may be related to the inhibition of caspase-3 activity and anti-apoptotic effects.