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目的探讨奥氮平对Aβ淀粉样蛋白25-35(Aβ_(25-35))诱导PC12细胞损伤的保护作用。方法将PC12细胞随机分为对照组、模型组、低中高3个剂量实验组。模型组和实验组用10μmol·L~(-1)Aβ_(25-35)诱导PC12细胞,构建氧化应激模型。低、中、高剂量实验组分予以10,30,100μg·mL~(-1)奥氮平培养,对照组和模型组予以不含任何药物的培养基。用噻唑蓝法检测细胞活力,用Hoechst染色法检测细胞凋亡情况,用分光光度法检测Caspase 3活性,用Western blot法检测B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)含量及磷脂酰肌醇3激酶/蛋白激酶B(PI3K/AKT)通路的激活状况。结果对照组、模型组和低中高3个剂量实验组的细胞活力分别为(100.00±9.35)%,(58.46±5.59)%,(67.43±6.52)%,(76.31±7.60)%,(89.21±8.85)%;Caspase 3活性分别为(1.00±0.12),(3.48±0.35),(2.85±0.28),(2.03±0.20),(1.34±0.13)U·mg~(-1);Bcl-2表达量分别为(0.68±0.06),(0.24±0.02),(0.38±0.04),(0.63±0.06),(0.69±0.07);Bax表达量分别为(0.27±0.02),(0.82±0.08),(0.63±0.06),(0.43±0.04),(0.30±0.03);PI3K表达量分别为(0.89±0.08),(0.22±0.02),(0.34±0.03),(0.50±0.05),(0.54±0.05);p-AKT表达量分别为(2.32±0.22),(0.28±0.02),(0.27±0.02),(1.88±0.18),(2.00±0.20);p-CREB表达量分别为(2.79±0.24),(0.18±0.01),(0.14±0.01),(0.47±0.04),(0.50±0.05);其中模型组的上述指标与对照组比较,差异均均有统计学意义(均P<0.01),低中高3个剂量实验组的细胞活力、Caspase 3活性、Bcl-2、Bax及PI3K表达量与模型组比较,差异均有统计学意义(均P<0.01),中、高剂量组p-AKT及p-CREB表达量与模型组比较,差异均有统计学意义(P<0.01)。结论奥氮平对Aβ_(25-35)诱导PC12细胞损伤具有显著的保护作用,可能与激活PI3K/AKT信号通路有关。
Objective To investigate the protective effect of olanzapine on PC12 cells induced by Aβ 25-35 (Aβ 25-35). Methods PC12 cells were randomly divided into control group, model group, low, medium and high dose three experimental groups. Model group and experimental group induced PC12 cells with 10μmol·L -1 Aβ 25-35 to establish oxidative stress model. Low, medium and high doses of experimental components were administered olanzapine at 10, 30 and 100 μg · mL -1, while control and model groups were given medium without any drug. Cell viability was detected by thiazolyl blue staining. Cell apoptosis was detected by Hoechst staining. Caspase 3 activity was detected by spectrophotometry. Bcl-2, Bcl-2 related protein X (Bax) and phosphoinositide 3 kinase / protein kinase B (PI3K / AKT) pathway activation status. Results The cell viability in control, model and low, medium and high dose groups were (100.00 ± 9.35)%, (58.46 ± 5.59)%, (67.43 ± 6.52)%, (76.31 ± 7.60)%, (89.21 ± 8.85)% respectively. The activities of Caspase 3 were (1.00 ± 0.12), (3.48 ± 0.35), (2.85 ± 0.28), (2.03 ± 0.20), (1.34 ± 0.13) U · mg -1 The expression levels of Bax were (0.68 ± 0.06), (0.24 ± 0.02), (0.38 ± 0.04), (0.63 ± 0.06), (0.69 ± 0.07) , (0.63 ± 0.06), (0.43 ± 0.04) and (0.30 ± 0.03), respectively. The PI3K expression levels were 0.89 ± 0.08, 0.22 ± 0.02, 0.34 ± 0.03, 0.50 ± 0.05, ± 0.05). The expression of p-AKT was (2.32 ± 0.22), (0.28 ± 0.02), (0.27 ± 0.02), (1.88 ± 0.18) and ± 0.24), (0.18 ± 0.01), (0.14 ± 0.01), (0.47 ± 0.04) and (0.50 ± 0.05), respectively. The differences between the above indexes in the model group and the control group were statistically significant (P < 0.01). Compared with model group, the cell viability, Caspase 3 activity, Bcl-2, Bax and PI3K expression in low, medium and high dose groups were significantly different (all P <0.01) The expression of p-AKT and p-CREB was lower than that of the model group It was statistically significant (P <0.01). Conclusion Olanzapine has a significant protective effect on PC12 cells induced by Aβ 25-35, which may be related to the activation of PI3K / AKT signaling pathway.