摘要
目的:观察知母总皂苷对β淀粉样蛋白(β-amyloid,Aβ)Aβ25-35诱导的PC12细胞中抗凋亡基因Bcl-2表达的影响。方法:培养PC12细胞,分为6组:正常组、模型组、知母低、中、高剂量组、盐酸多奈哌齐组。模型组加入终浓度为20μmol.L-1Aβ25-35的培养基,作用48 h;知母低、中、高剂量组和盐酸多奈哌齐分别加入含有低、中、高剂量知母总皂苷(5,10,20 mg.L-1)及盐酸多奈哌齐(1μmol.L-1)和Aβ25-35(终浓度为20μmol.L-1)的培养基共同作用于PC12细胞48 h。然后采用MTT检测各组细胞活力的变化;采用RT-PCR检测各组细胞中Bcl-2的mRNA表达变化;采用Western blotting技术检测各组细胞中Bcl-2蛋白的表达变化。结果:和正常组比较,模型组细胞活力降低(P<0.01);和模型组比较,高、中、低剂量知母总皂苷能显著升高PC12细胞活力(P<0.05);和正常组比较(0.476±0.072,0.14±0.02),模型组抗凋亡基因Bcl-2的mRNA(0.316±0.026)和蛋白(0.08±0.01)表达水平降低(P<0.01);和模型组比较,高、中、低剂量知母总皂苷能显著性升高抗凋亡基因Bcl-2的mRNA(0.447±0.016,0.465±0.043,0.472±0.023)和蛋白(0.17±0.01,0.19±0.02,0.13±0.01)的表达水平(P<0.05)。结论:知母总皂苷能升高Aβ25-35诱导的PC12细胞的活力降低,其机制可能是升高Bcl-2的表达水平。
Objective： To investigate the elevating expression of Bcl-2 by anemarrhena total saponins on beta-amyloid（Aβ）-induced PC12 cell injury.Method： PC12 cells were randomly divided into 6 group as following： normal group,model group,anemarrhena total saponins groups（5,10,20 mg.L-1）,the donepezil HCl group.Excluding the normal group,the PC12 cells in others groups were exposed to DMEM containing betaamyloid（20 μmol.L-1） and anemarrhena total saponins groups（5,10,20 mg.L-1,the donepezil HCl（1 μmol.L-1）.The mRNA and protein expression level of the Bcl-2 were detected using semi-quantitative reverse transcriptase PCR（RT-PCR） and western blot respectively.The cell survival were measured by MTT methods.Result： Anemarrhena total saponins significantly attenuated Aβ-induced PC12 cell injury,and elevated the mRNA and protein expression of Bcl-2,compared with model group（P ＆lt; 0.05）.Conclusion： Anemarrhena total saponins can inhibit PC12 cell injury induced by Aβ25-35,and the mechanism maybe involving in increasing,the expression levels of Bcl-2 mRNA and protein.
出处
《中国实验方剂学杂志》
CAS
北大核心
2013年第12期208-211,共4页
Chinese Journal of Experimental Traditional Medical Formulae
