氯胺酮上调GSK-3β活性加重Aβ_(25-35)诱导的大鼠PC12细胞Tau蛋白过度磷酸化  

Ketamine aggravates Tau hyperphosphorylation by up-regulating activity of GSK-3β in Aβ_(25-35 )induced PC12 cells

作　　者：许杰[1] 张蕊[2] 岳云[1] 左萍萍[3] 杨楠[3] 纪超[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]中国医学科学院北京协和医学院基础医学研究所药理室,北京100005

出　　处：《基础医学与临床》2011年第5期509-514,共6页Basic and Clinical Medicine

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

摘　　要：目的探讨氯胺酮对凝聚态Aβ25-35诱导的大鼠肾上腺嗜铬细胞瘤细胞(PC12)Tau蛋白过度磷酸化的影响及可能作用的机制。方法将培养的PC12细胞随机分为对照组(C)、10μmol/L Aβ25-35(A组)、1 mmol/L氯胺酮(K组)、氯胺酮和Aβ25-35(AK组),作用时间均为6 h。MTT比色法测定细胞活力,蛋白免疫印迹检测Tau蛋白不同磷酸化位点和糖原合成激酶3β(GSK-3β)在Ser9位点的磷酸化(p-GSK-3βSer9)相对于GSK-3β的表达水平,用GSK-3β特异性抑制剂氯化锂(LiCl)干预。结果 AK组的细胞存活率显著低于其余组;Tau蛋白在Ser396、Ser404和Thr231位点的磷酸化水平显著高于A组(P<0.05)。LiCl可通过增加p-GSK-3βSer9的表达抑制上述变化。结论氯胺酮通过上调GSK3β的活性增加Aβ25-35诱导的大鼠PC12细胞Tau蛋白过度磷酸化。Objective To find potential effect of ketamine can induce Tau hyperphosphorylation by increasing activity of glycogen synthase kinase-3β（GSK-3β） in Aβ25-35 induced PC12 cells.Methods We performed these studies in cultured rat pheochromocytoma cells（PC12） and randomly allocated PC12 cells into four groups： control group（C）,10 μmol/L Aβ25-35（A）,1 mmol/L ketamine（K）,Aβ25-35 and ketamine（AK）.All groups were incubated for 6 hours.The cell viability was determined by MTT assay.Western blot was performed to observe the protein expression of Tau phosphorylation of different sites,GSK-3β and p-GSK-3βSer9.In addition,lithium chloride,inhibitor of GSK-3β,was administered to all above groups to investgate the changes of protein expression.Results Thecell survival rate was significantly decreased in AK group as compared with the others groups.The level of Tau phosphorylation in the sites of Ser396,Ser404 and Thr231 increased after intubation with ketamine and Aβ25-35.And these increases were accompanied by up-regulation of activity of GSK-3β.Lithium chloride attenuates these changes above all.Conclusion Ketamine increases Tau hyperphosphorylation by up-regulating activity of GSK-3β in Aβ25-35 induced PC12 cells.

关 键 词：氯胺酮 TAU蛋白 PC12细胞 β淀粉样蛋白多肽片段25-35 GSK-3Β 

分 类 号：R614.1[医药卫生—麻醉学]