转染NEP基因对Aβ(下标25-35)诱导神经元凋亡的保护作用
张亚伦, 丁岩, 邵世滨, 杨玲玲, Kai Ren, 段珊, 曾季平, 崔行
Abstract
张亚伦, 丁岩, 邵世滨, 杨玲玲, Kai Ren, 段珊, 曾季平, 崔行
Abstract
Objective To explore the protective effects of transfection of the Neprilysin gene on primary cells against apoptosis induced by Aβ(subscript 25-35). Methods Primary cortical neurons of fetal Wistar rats in vitro were cultured and the apoptosis cell model was induced by Aβ(subscript 25-35). The NEP genes were transiently transfected into neurons. Changes of cell apoptosis were determined by MTT assay and flow cytometric DNA analysis. The expression of target mRNA and proteins was determined by RT-PCR and Western blot. Results Exposure of primary neurons to Aβ(subscript 25-35) resulted in changes of cell apoptosis. The MTT assay showed that cell activity remarkably decreased (P<0.01) and cytometric DNA analysis showed an increase in the apoptosis rate. While in the neurons transfected with NEP genes, there were no such remarkable changes when treated with Aβ(subscript 25-35). The suppression effect of the NEP gene on APP expression was shown by RT-PCR and Western blot, and the expression of Bax was decreased while that of Bcl-xl was increased after transfection. Conclusion Transfection of the NEP gene can protect primary neurons from apoptosis induced by Aβ(subscript 25-35).
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Objective To explore the protective effects of transfection of the Neprilysin gene on primary cells against apoptosis induced by Aβ(subscript 25-35). Methods Primary cortical neurons of fetal Wistar rats in vitro were cultured and the apoptosis cell model was induced by Aβ(subscript 25-35). The NEP genes were transiently transfected into neurons. Changes of cell apoptosis were determined by MTT assay and flow cytometric DNA analysis. The expression of target mRNA and proteins was determined by RT-PCR and Western blot. Results Exposure of primary neurons to Aβ(subscript 25-35) resulted in changes of cell apoptosis. The MTT assay showed that cell activity remarkably decreased (P<0.01) and cytometric DNA analysis showed an increase in the apoptosis rate. While in the neurons transfected with NEP genes, there were no such remarkable changes when treated with Aβ(subscript 25-35). The suppression effect of the NEP gene on APP expression was shown by RT-PCR and Western blot, and the expression of Bax was decreased while that of Bcl-xl was increased after transfection. Conclusion Transfection of the NEP gene can protect primary neurons from apoptosis induced by Aβ(subscript 25-35).
Key concepts: Apoptosis, Transfection, Molecular biology, Western blot, MTT assay, Flow cytometry, Cell culture, Biology