肥大细胞激活对大鼠蛛网膜下腔出血后血脑屏障损伤的作用  

Effect of mast cell activation on the blood-brain barrier injury after subarachnoid hemorrhage in rats

作　　者：孔德训 王翔 张继勤 谭赢 向欣 Kong Dexun;Wang Xiang;Zhang Jiqin;Tan Ying;Xiang Xin(Clinical Medical College of Guizhou Medical University,Guiyang 5504,China;Department of Anesthesiology,Guizhou Provincial People's Hospital,Guiyang 550002,China;Department of Neurosurgery,Guizhou Provincial People's Hospital,Guiyang 550002,China;Department of Neurosurgery,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China)

机构地区：[1]贵州医科大学临床医学院,贵阳550004 [2]贵州省人民医院麻醉科,贵阳550002 [3]贵州省人民医院神经外科,贵阳550002 [4]贵州医科大学附属医院神经外科,贵阳550004

出　　处：《中华实验外科杂志》2024年第9期1972-1975,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(82360376、82360482、81660239)。

摘　　要：目的 探讨肥大细胞(MCs)激活对大鼠蛛网膜下腔出血(SAH)后血脑屏障(BBB)损伤的作用及其机制.方法 采用血管内穿孔法制备SAH模型.将80只成年雄性Sprague-Dawley大鼠按随机数字表法分为8组,每组10只.另外66只大鼠按数字表法随机分为sham组、SAH组和色苷酸钠(Cro)组,每组22只.Cro组术后立即经腹腔注射25 mg/kg Cro,SAH组注射等体积0.9%NaCl.术后8h进行SAH评分、神经功能评分、脑含水量的测定、伊文思蓝染色和旷场实验.采用甲苯胺蓝染色和免疫组织化学染色检测MCs激活.利用蛋白免疫印记检测紧密连接蛋白(claudin-5)、闭锁小带蛋白-1(ZO-1)、闭锁蛋白(Occludin)、类胰蛋白酶(Tryptase)的表达.组间比较采用配对t检验.结果 大鼠蛛网膜下腔出血后8hMCs激活达高峰.SAH组伊文思蓝染色比sham组和Cro组较深.SAH组神经功能评分低于sham组和Cro组(12.90±0.89比18.00±0、15.80±1.26,t=9.75、2.47,P<0.05);SAH 组脑含水量高于 sham 组和 Cro 组(79.69±0.13 比 78.47±0.17、79.22±0.12,t=11.16、5.38,P<0.05).旷场实验SAH组大鼠进出中心区域次数、中心区域运动距离低于 sham 组和 Cro组(6.80±3.11 比 19.00±6.04、14.40±5.94,t=4.04、2.53,P<0.05),(0.44±0.09 比 0.96±0.20、0.86±0.20,t=4.33、4.12,P<0.05);SAH 组在中心区域时间高于sham 组和 Cro 组(153.40±44.42 比 93.60±15.11、52.20±20.63,t=2.85、4.62,P<0.05).SAH组 ZO-1 低于 sham 组和 Cro 组(0.25±0.10 比 0.63±0.05、0.51±0.08,t=5.91、3.45,P<0.05);SAH 组 Occludin 低于 sham 组和 Cro 组(0.18±0.01 比 0.67±0.07、0.46±0.07,t=12.40、7.21,P<0.05)、SAH 组 Claudin-5 低于 sham 组和 Cro 组(0.45±0.03 比 1.25±0.04、0.67±0.07,t=27.52、4.85,P<0.05);SAH 组 Tryptase 高于 sham 组和 Cro 组(0.60±0.01 比 0.33±0.01、0.42±0.01,t=28.13、20.31,P<0.05).结论 大鼠蛛网膜下腔出血后8 h MCs激活达高峰;MCs的激活促进了血脑屏障的破坏介导了早期的脑损伤,肥大细胞稳定剂Cro抑制了 MCs的激活Objective To investigate the effect and mechanism of mast cells(MCs)activation on blood-brain barrier(BBB)injury after subarachnoid hemorrhage(SAH)in rats.Methods The SAH mod-el was prepared by intravascular perforation.A total of 80 adult male Sprague-Dawley rats were divided into 8 groups of 10 rats in each group according to the random number table method.The other 66 rats were ran-domly divided into sham group,SAH group and cromolyn sodium(CRO)group according to the numerical table method,with 22 rats in each group.The CRO group was injected with 25 mg/kg CRO intraperitoneal-ly immediately after surgery,and the SAH group was injected with the same volume of normal saline.SAH score,neurological function score,brain water content measurement,Evans blue staining and open field test were performed 8 h after surgery.Toluidine blue staining and immunohistochemical staining were used to detect MCs activation.Western blotting was used to detect the expression of claudin-5,ZO-1,Occludin,and tryptase.The t-test was used for comparison between groups.The Paired t-tests were used for compari-sons between groups.Results The expression of tryptase and the activation of MCs peaked 8 h after sub-arachnoid hemorrhage in rats.Evans blue staining in the SAH group was darker than that in the sham group and the Cro group.The neurological function score of the SAH group was lower than that of the sham group and the CRO group(12.90±0.89 vs.18.00±0,15.80±1.26,t=9.75,2.47,P<0.05).The cere-bral water content of the SAH group was higher than that of the sham group and the CRO group(79.69±0.13 vs.78.47±0.17,79.22±0.12,t=11.16,5.38,P<0.05).The number of entries and exits in the SAH group and the distance of movement in the center were lower than those in the sham group and the CR0 group(6.80±3.11 vs.19.00±6.04,14.40±5.94,t=4.04,2.53,P<0.05),(0.44±0.09 vs.0.96±0.20,0.86±0.20,t=4.33,4.12,P<0.05).The central time in the SAH group was longer than that in the sham group and the CRO group(153.40±44.42 vs.93.60±15.11,52.20±20.63,

关 键 词：肥大细胞 虫蛛网膜下腔出血 早期脑损伤 血脑屏障 

分 类 号：R743.35[医药卫生—神经病学与精神病学]