【摘要】 目的研究ATP敏感的钾离子通道开放剂二氮嗪(diazoxide)预处理对Aβ1~42作用原代培养神经元N-甲基-D-天冬氨酸(NMDA)受体2B(NR2B)亚基蛋白表达的影响。方法原代培养大鼠皮层海马神经元并进行鉴定,将细胞随机分为对照组、单纯Aβ1~42干预组、二氮嗪预处理1h后Aβ1~42干预组(Aβ1~42+diazoxide)、单纯二氮嗪预处理组,并采用免疫印迹检测不同时间点(24 h、72 h)细胞NR2B亚基蛋白表达水平的变化。结果 Aβ1~42(2μmol/L)作用神经元24 h后,与对照组相比,单纯Aβ1~42干预组和Aβ1~42+diazoxide组的NR2B亚基蛋白表达均无明显改变。Aβ1~42作用神经元72 h后,与对照组比较,单纯Aβ1~42组NR2B亚基蛋白表达量显著升高(P<0.05);而与单纯Aβ1~42干预组相比,Aβ1~42+diazoxide组NR2B亚基蛋白表达明显降低(P<0.05)。结论 Aβ1~42作用原代培养神经元72 h,能够显著增加神经细胞NR2B亚基蛋白的表达量;同时,二氮嗪能拮抗Aβ1~42所引起的NR2B亚基蛋白表达量升高,提示二氮嗪可能通过NMDA受体通路影响Aβ1~42的细胞毒性作用。更多还原

【Abstract】 Objective To study the effects of pretreatment of ATP sensitive potassium channel opener diazoxide on neural NR2 B subunit protein expression induced by Aβ1 ~ 42. Methods The primary neurons were cultured and evaluated,then neurons were randomly divided into control group,Aβ1 ~ 42 treatment group,diazoxide pretreatment for 1 h + Aβ1 ~ 42group( Aβ1 ~ 42+ diazoxide) and simple diazoxide group,the protein expression level of NR2 B subunit was determined by Western blotting for different times( 24 h and 72 h). Results Being exposed to Aβ1 ~ 42 for 24 h,compared with the control group,the protein expression level of NR2 B subunit of both Aβ1 ~ 42 group and Aβ1 ~ 42+ diazoxide group have no significant difference. However,compared with control group,the protein expression level of NR2 B subunit of Aβ1 ~ 42 group significantly increases for 72 h( P < 0. 05); Compared with Aβ1 ~ 42 group,the protein expression level of NR2 B subunit of Aβ1 ~ 42+ diazoxide group significantly decreases( P < 0. 05). Conclusions Aβ1 ~ 42 could increase the protein expression level of NR2 B subunit in cultured primary neurons for 72 h,and ATP sensitive potassium channel opener diazoxide could inhibit the increasing of protein expression level of NR2 B subunit induced by Aβ1- 42,it is indicated that diazoxide might affect Aβ1 ~ 42-induced cytotoxicity via NMDA receptor pathway.更多还原