咪达唑仑对凝聚态Aβ25-35诱导的大鼠PC12细胞Tau蛋白过度磷酸化和凋亡蛋白的影响  

Effect of midazolam on Tau hyperphosphorylation and apoptosis proteins in cultured PC12 cell after Aβ_(25-35) induction

作　　者：许杰[1] 张蕊[2] 岳云[1] 左萍萍[3] 杨楠[3] 刘雁勇[3] 

机构地区：[1]首都医科大学附属北京朝阳医院麻醉科,北京100020 [2]潍坊医学院麻醉学系,山东潍坊261042 [3]中国医学科学院基础医学研究所,北京协和医学院药理室,北京100005

出　　处：《解剖学报》2011年第3期334-339,共6页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(30872425);山东省教育厅国内访问学者资助项目(BK2001132)

摘　　要：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 Midazolam is one of the most commonly used anesthetics in operating room and intensive care unit. However, the effects of midazolam on AD neuropathogenesis have not been well determined. Here, we set out to assess the effects of midazolam on Tau hyperphosphorylation and expression of apoptosis protein in beta-amyloid peptid%5.35 （Aβ25-35） treated PC12 cells. Methods The pheochromoeytoma cells（PC12） were randomly allocated into four groups： control group（C） ; 10 p, mol/L Aβ25-35 （A） ; 20μmol/L midazolam （M） ; Aβ25-35 and midazolam （MA） , above all groups were incubated for 6 hours. The cell viability was determined by MTT assay, Western blotting and immunocytochemical stain were performed to observe the protein expression of Bcl-2 family and Tau phosphorylation at different sites. Results Incubating PC12 cells with midazolam at different concentration with Aβ25-35 can decrease the number of living cells in a dose-dependent way. The level of Tau protein phosphorylation in the sites of Ser396 and Ser404 increased in MA group compared with A group （P 〈 0. 05） , and the Bax level of MA group significantly higher than A group （P 〈 0. 05 ）. Conclusion Midazolam aggravates the Tau hyperphosphorylation and expression of apoptosis protein in cultured PC12 ceils after Aβ25-35induction.

关 键 词：咪达唑仑 Bcl-2/Bax TAU蛋白 PC12细胞 β淀粉样蛋白多肽片段25~35 免疫印迹法 大鼠 

分 类 号：P614.1[天文地球—矿床学]