三种方法检测Aβ所致PAJU细胞凋亡

目的分析三种检测细胞凋亡方法在观察缺氧条件下Aβ所致PAJU细胞凋亡中的运用。方法将人神经母细胞瘤细胞分为常态组、Aβ干预组、缺氧组、缺氧+Aβ干预组。分别用MTT法、Hoechst 33258细胞核染色法、流式细胞术检测细胞凋亡。结果 MTT法结果显示,缺氧、Aβ42、Aβ42并缺氧处理组与对照组相比其存活率之间的差异具有统计学意义(P<0.05)。Hoechst 33258细胞核染色法:对照组的PAJU细胞,细胞中核浓染、边集及碎裂比例较少,经缺氧、Aβ42处理、缺氧+Aβ42共同处理后,细胞中核浓染、边集及碎裂比例较多;各处理组的凋亡核百分率与对照组相比其差异具有统计学意义(P<0.05);流式细胞仪检测结果:PAJU对照组与各处理组细胞凋亡率的差异具有统计学意义(P<0.05),缺氧组与Aβ组比较凋亡率无显著变化,其差异无统计学意义(P>0.05)。结论 Aβ42能引起PAJU细胞的凋亡,缺氧能促进其凋亡。

Three methods for detecting the apoptosis of PAJU cells induced by amyloid beta-protein

Objective To explore application of MTT,Hoechst 33258 staining and flow cytometry detecting the apoptosis of PAJU cells induced by amyloid beta-protein under hypoxia condition.Methods PAJU cells were divided into ①normal group: 50 ml/L culturing PAJU cells in CO2 box for 96 h;②Aβ group:giving PAJU cells 1.0 nmol/L aged Aβ42,then culturing them for 96 h;③hypoxia group: culturing PAJU cells in 1%～2% O2,5% CO2,93% N2 box for 96 h;④Aβ+hypoxia group: giving PAJU cells 1.0 nmol/L aged Aβ42,culturing them in 1%～2% O2,5% CO2,93% N2 box for 96 h.Apoptosis of PAJU cells were observed by MTT,Hoechst 33258 staining and flow cytometry methods respectively.Results The number of PAJU cells of Aβ,hypoxia and Aβ+ hypoxia groups were decreased,and the activity of them were decreased compared with controlled ones in MTT,Hoechst 33258 staining and flow cytometry respectively.There was statistically significance between control and experimental groups(P<0.05),and there was no statistically significance between Aβ group and hypoxia groups in apoptosis rate(P>0.05).Conclusions Aβ42 could cause the apoptosis of PAJU cells,hypoxia could enhance the apoptosis.