奥氮平抑制Aβ淀粉样蛋白25-35诱导PC12细胞损伤的保护作用

Protective effect of olanzapine on amyloid β_(25-35) induced PC12 cells injury

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 explore the protective effect of olanzapine on amyloid β25-35 （Aβ25-35） induced PC12 cells injury. Methods The cells were divided into normal group , model group and low -dose, middle-dose, high -dose test groups. The model group and test group were constructed oxidative stress model with 10 μmol · L^-1 Aβ25-35. The low- dose, middle- dose, high- dose test groups were given 10, 30, 100 μmol · L^-1 olanzapine, respectively. And control and model groups were given blank media without drug. The viability of PC12 cells, cell apoptosis and activity of Caspase 3 were measured by MTT assay, Hoechst staining and spectrophotometry, respectively. The B -eel1 lymphoma -2 （Bcl -2）, Bcl -2 associated X protein （Bax） and the activation of phosphatidylinositol 3 kinase / protein kinase B （PI3K/AKT） were measured by Western blot. Results In the control group, model group and low- dose, middle - dose, high - dose test groups, the viability of PC12 cells were （ 100.00 ± 9. 35 ） %, （ 58.46 ± 5.59 ） %, （67.43±6.52）%, （76.31 ±7.60）%, （89.21 ±8.85）%; the activity 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 of Bel -2 were （0.68 ±0.06）, （0.24±0.02）, （0.38 ±0.04）, （0.63 ±0.06）, （0.69 ±0.07）; the expression of Bax were （0.27±0.02）, （0.82±0.08）, （0.63±0,06）, （0.43±0.04）, （0.30 ±0.03）; the expression of PI3K were （0.89±0.08）, （0.22±0.02）, （0.34±0.03）, （0.50±0.05）, （0.54±0.05）; the expression ofp-AKTwere （2.32 ±0.22）, （0.28 ±0. 02）, （0. 27 ±0.02）, （1.88 ±0. 18）, （2. 00 ±0. 20） ; the expression ofp-CREB were （2.79±0.24）, （0.18 ±0.01）, （0.14 ±0.01）, （0.47 ±0.04）, （0.50 ±0.05）. Compared with the control group, the those indexes in model group had a significant difference （ P 〈 0. 01 ） ; the differences on viability of PC12 cells, activity of Caspase 3, expression of Bax, Bcl - 2 and PI3K in low - dose, middle - dose, high - dose test groups and model group were statistically significant （ P 〈 0. 01 ） . Also, compared with model group, the differences of p - AKT and p - CREB in middle - dose, high - dose test groups were significant （ P 〈 0. 01 ）. Conclusion Olanzapine has a definitive protect efficacy for Aβ25-35 induced PC12 cells injury, which might be related to activating PI3K/AKT signal pathway.