K_(ATP)通道开放剂对氧糖剥夺诱导的SH-SY5Y细胞凋亡及AIF信号通路的影响  

Effects of ATP sensitive potassium channel opener on SH-SY5Y cells apoptosis and AIF signaling pathway induced by oxygen and glucose deprivation

作　　者：王占强[1] 张鸿[1] 何平平[1] 

机构地区：[1]中国医科大学附属盛京医院神经内科,辽宁沈阳110004

出　　处：《解剖科学进展》2015年第2期184-187,191,共5页Progress of Anatomical Sciences

基　　金：辽宁省科学技术计划项目(2012225069);沈阳市科学技术计划(F12-277-1-02);辽宁省"百千万人才工程"资助项目(2011921041)

摘　　要：目的观察KA TP通道开放剂对氧糖剥夺诱导SH-SY5Y细胞凋亡的影响,并进一步探讨K ATP通道开放剂对凋亡诱导因子(apoptosis inducing factor,AIF)信号通路的影响。方法取传代后3d的SH-SY5Y细胞,分为A组(正常对照组)、B组(氧糖剥夺组)、C组(二氮嗪处理组)、D组(二氮嗪+5-羟葵酸处理组)。采用MTT方法检测细胞活力,应用流式检测细胞凋亡,应用Western-blot及免疫荧光染色检测AIF蛋白表达变化及移位。结果与正常对照组细胞相比,氧糖剥夺后细胞活力明显降低,100及200μmol/L的二氮嗪处理以后细胞活力明显升高,200μmol/L二氮嗪升高细胞活力更明显。与正常对照组细胞相比,氧糖剥夺后细胞凋亡率明显增高,二氮嗪处理以后细胞凋亡率明显下降。A,B,C,D四组AIF蛋白总量表达无显著性的差异(P>0.05);在正常对照组,AIF蛋白以胞浆表达为主,氧糖剥夺导致AIF发生了明显的细胞核转移,二氮嗪处理显著减少氧糖剥夺诱导的AIF发生的细胞核转移。结论 KA TP通道开放剂二氮嗪能明显增加氧糖剥夺后SH-SY5Y细胞活力,减少细胞凋亡,减少氧糖剥夺诱导的AIF发生的细胞核转移。Objective To observe the effects of KATP channel openers on OGD induced SH-SY5 Y cells apoptosis and apoptosis inducing factor（AIF） signaling pathway. Methods 3d passaged SH-SY5 Y cells were divided into A group（normal control）, B group（OGD group）, C group（diazoxide treatment group） and D group（diazoxide＋5-HD treatment group）. Cell viability was detected by MTT method, neuronal apoptosis was detected by Heochst 33342/PI double-dyed flow cytometry, AIF protein expression and shift were detected by Western-blot and immunofluorescence staining. Results The cell viability decreased significantly after OGD compared with normal control, but increased significantly after 100 and 200μmol/L diazoxide treatment. The cells apoptosis rate increased significantly after OGD treatment, but decreased significantly after diazoxide treatment compared with normal control group. The expression level of AIF protein had no significant differences for A, B, C and D groups, AIF proteins were mainly expressed in the cytoplasm in normal control group, with obvious nuclear transfer after OGD treatment, but reduced significantly after diazoxide treatment. Conclusion KATP channel opener can significantly increase SH-SY5 Y cell viability, reduce cell apoptosis and decrease AIF nuclear transfer after OGD.

关 键 词：ATP敏感性钾通道 SH-SY5Y细胞 二氮嗪 AIF 5-HD 

分 类 号：Q255[生物学—细胞生物学]