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目的 通过形态学和生化学指标观察APP17肽对Aβ2 5 3 5诱发SH SY5Y细胞凋亡是否有保护作用 ,并对其保护作用机制作进一步的探讨。方法 选用细胞形态观察、半定量LM PCRDNAladderAssay和流式细胞仪定量测定细胞凋亡率的方法观察APP17肽是否对Aβ2 5 3 5诱发SH SY5Y细胞凋亡有保护作用 ;通过测定细胞MTT代谢率 ,共聚焦显微镜观察Aβ2 5 3 5和APP17肽对细胞内过氧化物含量、线粒体膜电位的影响以及WesternBlotting观察细胞内凋亡诱导因子AIF及核转录因子NF κB的表达情况 ,探讨了APP17肽的保护作用机制。结果 APP17肽可以明显改善Aβ2 5 3 5造成的细胞形态损伤 ,显著降低细胞凋亡比率 ;与Aβ2 5 3 5损伤组相比 ,APP17肽预孵育能明显提高细胞MTT代谢率 ,稳定线粒体膜电位 ,减少AIF的表达 ;有效降低Aβ2 5 3 5引起的细胞内过氧化物含量增高 ;并且使具有神经保护作用的核转录因子NF κB表达增加。结论 APP17肽可以稳定线粒体功能、降低细胞内过氧化物含量 ,激活NF κB并使其持续表达 ,对Aβ2 5 3 5诱导的SH SY5Y细胞凋亡有比较明显的保护作用。
OBJECTIVE: To observe whether APP17 peptide has a protective effect on the apoptosis of SH SY5Y cells induced by Aβ2 5 3 5 by morphological and biochemical indexes and to further explore its protective mechanism. Methods Cell morphology, semi-quantitative LM PCRDNAladderAssay and flow cytometry were used to determine the apoptotic rate. To observe whether APP17 peptide has a protective effect on the apoptosis of SH SY5Y cells induced by Aβ2 5 3 5. MTT metabolic rate The effects of Aβ2 5 3 5 and APP17 peptide on intracellular peroxide content and mitochondrial membrane potential were observed under a confocal microscope. The expression of AIF and NF κB in cells were observed by Western Blotting. The protective effect of APP17 peptide mechanism. Results APP17 peptide could significantly improve cell morphological damage induced by Aβ2 5 3 5 and significantly reduce the apoptosis rate. Compared with Aβ2 5 3 5 injury group, pretreatment with APP17 peptide significantly increased the MTT metabolic rate, mitochondrial membrane potential, Reduce the expression of AIF; effectively reduce the Aβ2 5 3 5-induced increase in intracellular peroxide content; and increase the neuroprotective effect of nuclear transcription factor NF κB expression. Conclusion APP17 peptide can stabilize mitochondrial function, decrease intracellular peroxide content, activate NF κB and persistently express it, and have a significant protective effect on apoptosis of SH SY5Y cells induced by Aβ2 5 3 5.