作　　者： (陈兴泳）; (张旭）; (雷惠新）; (廖之君）; (汪银洲）;

机构地区： 福建医科大学省立临床医学院、福建省立医院神经内科,福州350001

出　　处： 《中国临床药理学杂志》 2017年第17期1687-1689,1705,共4页

摘　　要： 目的观察RNA干扰免疫蛋白酶体亚基低分子重量蛋白2(LMP2)对脑缺血再灌注大鼠血脑屏障结构相关蛋白闭合蛋白(Occludin)和闭锁小带蛋白(ZO^(-1))表达的影响。方法线栓法制作SD大鼠大脑中动脉阻塞再灌注(MCAO)模型,脑缺血1 h再灌注72 h。按照体重将SD大鼠随机分为3组:假手术组、实验组和对照组,每组10只。在MCAO术前1 h,实验组和对照组分别立体定位地脑内注射慢病毒载体LMP2-shRNA和阴性对照shRNA液体。以免疫印迹法分析Occludin、ZO^(-1)蛋白表达,用伊文思蓝(EB)示踪法监测血脑屏障损伤程度。结果 Occludin蛋白与ZO^(-1)蛋白的灰度值:假手术组、对照组和实验组分别1.76±0.09,1.37±0.12;0.24±0.07,0.26±0.07;0.83±0.11,0.74±0.09,与假手术组比较,对照组差异均有统计学意义(均P<0.001);与对照组比较,实验组差异也均有统计学意义(均P<0.001)。脑组织EB含量:假手术组、对照组和实验组分别(3.32±0.14),(11.41±0.21),(6.23±0.28)μg·g^(-1),与假手术组比较,对照组差异有统计学意义(P<0.001);与对照组比较,实验组差异也有统计学意义(P<0.001)。结论沉默LMP2基因表达可显著改善大鼠脑缺血后血脑屏障透通性。 Objective To investigate the effects of RNA interference targeting immunoproteasome subunit low molecular weight protein 2 （LMP2） on expression of Occludin and zonula occludens （ ZO - 1 ） of blood brain barrier （BBB） after cerebral ischemia/reperfusion in a rat model. Methods The middle cerebral artery occlusion（ MCAO ）was in- duced using the intraluminal suture occlusion technique in SD rats. Reperfusion of cerebral blood flow was allowed by gently removing the monofilament after 1 hour ischemia, followed by 72 h reperfusion. Rats were randomly assigned into three groups： sham - operated group, experimental group and control group （t = 10 in each group ）. The preparations of lentivirus vector targeting LMP2 short hairpin RNA （shRNA） and carrying scrambled shRNA were infused stereotactically into the ipsilateral hemispheric region 1 h before MCAO in the experimental group and control group, respectively. The expressions of Oeeludin and ZO - 1 protein in the brain were detected by Western blot, respectively. BBB permeability was detected by Evans Blue （EB）leakage. Results The expression of gray value of Occludin and ZO - 1 ：it in sham - operated group, control group and experimental group were 1.76 ±0. 09,1.37 ± 0. 12 ;0. 24 ± 0. 07,0. 26 ± 0. 07 ;0. 83 ± 0. 11,0. 74 ± 0. 09. Compared with sham - operated group, the expression of Occludin and ZO - 1 in control group were reduced significantly （ all P 〈0. 001）. Compared with control group, the expression of Occludin and ZO- 1 in experimental group were also reduced significantly （all P 〈0. 001 ）. The contents of EB：it in sham -operated group, control group and experimental group were （ 3.32 ± 0. 14 ） , （ 11.41 ± 0. 21 ） , （ 6.23 ± 0. 28 ）μg·g-（-1）. Compared with sham - operated group, the contents of EB in control group were reduced significantly （all P 〈 0. 001 ）. Compared with control group, the contents of EB in experimental group were also reduced significantly （ all P 〈 0. 001 �

关 键 词： 免疫蛋白酶体 干扰低分子重量蛋白 血脑屏障 脑缺血