白蛋白对老年大鼠全脑缺血再灌注后G蛋白偶联受体124表达和血脑屏障的影响  

Effects of albumin on the expression of G-protein coupled receptor 124 and blood-brain barrier after global cerebral ischemia-reperfusion in elderly rats

作　　者：唐跃东[1,2,3] 谢启亮 汪兰曌 杨斐宇 张峰 周小勇[1,2,3] 申捷 Tang Yuedong;Xie Qiliang;Wang Lanzhao;Yang Feiyu;Zhang Feng;Zhou Xiaoyong;Shen Jie(Center of Emergency and Critical Medicine,Jinshan Hospital Affiliated to Fudan University,Shanghai,201508,P.R.China;Key Laboratory of Chemical Injury,Emergency and Critical Medicine of Shanghai Municipal Health Commission,Shanghai,201508,P.R.China;Research Center for Chemical Injury,Emergency and Critical Medicine of Fudan University,Shanghai,201508,P.R.China)

机构地区：[1]复旦大学附属金山医院急危重病中心,上海201508 [2]上海市卫生健康委员会化学伤害急危重病医学重点实验室,上海201508 [3]复旦大学化学伤害与急危重病研究中心,上海201508

出　　处：《老年医学与保健》2022年第6期1245-1251,共7页Geriatrics & Health Care

基　　金：上海市金山区卫生健康委员会科研课题面上项目(JSKJ-KTMS—2019-04);上海市公共卫生体系建设三年行动计划(2020—2022年)重点学科建设计划基金(GWV-10.1-XK26)。

摘　　要：目的研究人血白蛋白(HSA)对老年大鼠全脑缺血/再灌注后血脑屏障通透性和G蛋白偶联受体124(GPR124)表达的影响。方法选用老年健康雄性Wistar大鼠共108只,按随机法分成3组:假手术组(Sham)、全脑缺血再灌注模型组(GCI/R)和人血白蛋白治疗组(GCI/R+HSA),每组36只。应用改良Pulsinelli四血管阻断法(4-VO)构建全脑缺血再灌注大鼠模型(GCI/R),短暂缺血15 min后再灌注。再灌注2 h后,经颈外静脉注射2.5 g/kg的HSA。再灌注后6、24、48 h,用伊文思蓝(EB)定性法分别检测血脑屏障的通透性。采用Western Blot(WB)、实时荧光定量PCR(qRT-PCR)和免疫荧光分析(IF)方法检测GPR124表达的变化。结果脑组织中的伊文思蓝(EB)在再灌注后开始增加,这种增加趋势可以通过HSA的干预来减轻。免疫荧光分析(IF)显示,GCI/R组GPR124的平均荧光强度在全脑缺血再灌注后24 h显著高于Sham组(P<0.01);与GCI/R组相比,GCI/R+HSA组GPR124的平均荧光强度进一步增加,表明GPR124表达增加,具有显著性差异(P<0.05)。WB和qRT-PCR检测到的GPR124蛋白和mRNA的表达趋势与IF结果基本一致。结论HSA能减轻大鼠全脑缺血/再灌注损伤和血脑屏障的破坏,促进GPR124在脑组织中的表达。GPR124可能在HSA的脑保护机制中发挥重要作用。Objective To investigate the effects of human serum albumin(HSA)on the permeability of blood-brain barrier(BBB)and the expression of G-protein coupled receptor 124(GPR124)after global cerebral ischemia/reperfusion(GCI/R)in elderly rats.Methods 108 elderly male Wistar rats were randomly divided into 3 groups:sham operation group,global cerebral ischemia/reperfusion(GCI/R)group and HSA treatment(GCI/R+HSA)group,with 36 ones in each group.The modified Pusinelli four-vessel occlusion method(4-VO)was used to establish a rat model of global cerebral ischemia/reperfusion(GCI/R),which was reperfusion after transient ischemia for 15 min.Two hours after reperfusion,2.5 g/kg HSA was injected through external jugular vein.The permeability of BBB was detected by Evans Blue(EB)qualitative method at 6,24,48 h after reperfusion,respectively.Western blot(WB),quantitative real-time PCR(qRT-PCR)and immunofluorescence(IF)analysis were used to detect the changes of GPR124 expression.Results EB in brain tissue began to increase after reperfusion,and this increasetrend can be diminished by the administration of HSA.IF assay demonstrated that the mean fluorescence intensity of GPR124 in the GCI/R group was significantly higher than that in the sham group 24 h after global cerebral ischemia reperfusion(P<0.01).Compared with GCI/R group,the mean fluorescence intensity of GPR124 in the GCI/R+HSA group was further increased,indicating that the expression of GPR124 was increased(P<0.05).The expression trends of GPR124 proteins and mRNA detected by WB and qRT-PCR were basically consistent with the results of IF.Conclusion HSA can alleviate the global cerebral ischemia/reperfusion injury and the damage of BBB in rats,and promote the expression of GPR124 in brain tissue.GPR124 may play an important role in the brain protection mechanism of HSA.

关 键 词：老年 大鼠 全脑缺血/再灌注损伤 血脑屏障 人血白蛋白 G蛋白偶联受体124 

分 类 号：R743[医药卫生—神经病学与精神病学]