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目的探讨咪达唑仑对凝聚态Aβ25～35诱导的大鼠PC12细胞Tau蛋白过度磷酸化的影响及凋亡相关蛋白表达的变化。方法将培养的PC12细胞随机分为4组:对照组(C组);10μmol/L Aβ25～35(A组);20μmol/L咪达唑仑(M组);咪达唑仑和Aβ25～35(MA组),作用时间均为6小时。四甲基偶氮唑盐(MTT)比色法测定细胞活力;用免疫印迹法和免疫细胞化学染色,观察咪达唑仑对Aβ25～35诱导的PC12细胞不同磷酸化位点Tau蛋白的表达、凋亡相关蛋白(Bcl-2、Bax)的表达和PC12细胞形态的变化。结果咪达唑仑作用于凝聚态Aβ25～35诱导的PC12可使凋亡相关蛋白Bax表达增高,Bcl-2表达降低。Tau蛋白Ser396、Ser404位点磷酸化水平显著增高(P<0.05)。结论咪达唑仑可增强凝聚态Aβ25～35诱导的大鼠PC12细胞Tau蛋白的过度磷酸化和细胞凋亡蛋白表达的增加。
Objective To investigate the effect of midazolam on Tau hyperphosphorylation in rat PC12 cells induced by condensed Aβ25-35 and the changes of apoptosis related protein expression. Methods The cultured PC12 cells were randomly divided into 4 groups: control group (group C), 10μmol / L Aβ25-35 (group A), 20μmol / L midazolam (M group), midazolam and Aβ25-35 (MA group), the role of time are 6 hours. MTT assay was used to measure cell viability. Western blotting and immunocytochemical staining were used to observe the effect of midazolam on the expression of Tau protein at different phosphorylation sites in PC12 cells induced by Aβ25-35. Apoptotic Related Protein (Bcl-2, Bax) Expression and PC12 Cell Morphology. Results Midazolam induced the expression of Bax and Bcl-2 in PC12 induced by condensed Aβ25-35. Tau protein Ser396, Ser404 phosphorylation level was significantly higher (P <0.05). Conclusion Midazolam can increase the expression of Tau hyperphosphorylation and apoptosis protein in rat PC12 cells induced by condensed Aβ25-35.