基于SIRT1/FoxO1信号通路探讨高压氧对脑缺血再灌注损伤大鼠血脑屏障的影响  

Effects of hyperbaric oxygen on the blood-brain barrier via the SIRT1/FoxO1 signaling pathway after cerebral ischemia and reperfusion

作　　者：佟雪 张倩茹[1] 赵红[1] 陈学新[2] Tong Xue;Zhang Qianru;Zhao Hong;Chen Xuexin(Functional Experimentation Center,College of Basic Medical Science,China Medical University,Shenyang 110122,China;Hyperbaric Oxygen Department,The First Affiliated Hospital of China Medical University,Shenyang 110001,China)

机构地区：[1]中国医科大学基础医学院机能实验中心,沈阳110122 [2]中国医科大学附属第一临床医院高压氧科,沈阳110122

出　　处：《中华物理医学与康复杂志》2022年第1期13-17,共5页Chinese Journal of Physical Medicine and Rehabilitation

基　　金：辽宁省科技厅博士启动基金资助项目(20101149)。

摘　　要：目的基于沉默信息调节因子1/叉头转录因子O1(SIRT1/FoxO1)信号通路探讨高压氧(HBO)对脑缺血再灌注(CIR)大鼠血脑屏障(BBB)的保护作用。方法选取雄性Wistar大鼠40只,按照随机数字表法将其分成假手术组(Sham组)、模型组(CIR组)、CIR+HBO组、CIR+HBO+SIRT1抑制剂(EX527)组,每组10只。采用改良线栓法建立大鼠右侧大脑中动脉栓塞模型,Sham组大鼠不进行结扎和线栓导入,CIR+HBO组和CIR+HBO+EX527组予以HBO干预,CIR+HBO+EX527组在此基础上再予以EX527处理。CIR+HBO组和CIR+HBO+EX527组于再灌注1 h、9 h、21 h、45 h、69 h进入HBO舱,期间行HBO治疗5次。在处死动物前1 h,经尾静脉注射2%伊文思兰(EB),采用比色法、逆转录-聚合酶链(RT-PCR)和Western blot法分别检测再灌注72 h大鼠海马区脑组织EB含量,SIRT1、FoxO1、ZO-1、Occludin、Claudin-5 mRNA及蛋白表达变化。结果CIR 72 h,CIR组、CIR+HBO组和CIR+HBO+EX527组大鼠海马区脑组织EB含量均较Sham组明显增加(P<0.05),CIR+HBO+XE527组大鼠海马区脑组织EB的含量较CIR+HBO组显著增加(P<0.05)。CIR 72 h,CIR组、CIR+HBO组和CIR+HBO+EX527组大鼠海马区脑组织SIRT1、FoxO1、ZO-1、Occludin、Claudin-5 mRNA表达及其相应蛋白表达较Sham组均显著降低(P<0.05),CIR+HBO+EX527组大鼠海马区脑组织SIRT1、FoxO1、ZO-1、Occludin、Claudin-5 mRNA表达及其相应蛋白表达较CIR+HBO组显著降低(P<0.05)。结论HBO可以通过SIRT1/FoxO1通路增加紧密连接蛋白的表达,从而在CIR损伤中对BBB起到保护作用。Objective To explore the effect of hyperbaric oxygen(HBO)on the blood-brain barrier via the silent information regulator 1(SIRT1)/Forkhead box O1(FoxO1)signaling pathway after cerebral ischemia and reperfusion using a rat model.Methods Forty Wistar rats were randomly assigned into sham,cerebral ischemia-reperfusion(CIR),CIR+HBO and CIR+HBO+EX527 groups,each of 10.The cerebral ischemia-reperfusion model was established in all groups except the sham group by right middle cerebral artery occlusion using the modified thread-occlusion method.The sham group was not ligated.Both the CIR+HBO and CIR+HBO+EX527 groups were given HBO 1,9,21,45 and 69 hours after the reperfusion.The CIR+HBO+EX527 group was additionally injected with 5mg/kg of EX527(a SIRT1inhibitor)peritoneally 4,12,24,48 and 72 hours after the reperfusion.Then 2%Evens blue(EB)was injected into the tail vein an hour before the rats were sacrificed.The content of EB and the expression of SIRT1,FoxO1,ZO-1,Occludin,Claudin-5 mRNA and their proteins were determined using spectrophotometry,reverse transcription-polymerase chain reactions and Western blotting.Results The average EB content of the hippocampal brain tissue from the CIR,CIR+HBO and CIR+HBO+EX527 rats was significantly greater than the Sham group′s average 72h after reperfusion.The average expression of SIRT1,FoxO1,ZO-1,Occludin and Claudin-5 mRNA and their proteins was significantly lower,with the CIR+HBO+EX 527 group′s average significantly lower than that of the CIR+HBO group.Conclusions HBO can increase the expression of tight junction protein via the SIRT1/FoxO1 pathway.It helps to protect the blood-brain barrier in CIR injury situations.

关 键 词：高压氧 血脑屏障 脑缺血再灌注 SIRT1 FOXO1 紧密连接蛋白 

分 类 号：R743.3[医药卫生—神经病学与精神病学]