RNA干扰低分子重量蛋白2基因对脑缺血大鼠血脑屏障的影响

发布时间：2018-04-11 08:30

本文选题：免疫蛋白酶体 + RNA干扰低分子重量蛋白　；参考：《中国临床药理学杂志》2017年17期

【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P0.001);与对照组比较,实验组差异也均有统计学意义(均P0.001)。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(P0.001);与对照组比较,实验组差异也有统计学意义(P0.001)。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。
[Abstract]:Objective To observe the effect of RNA interference immunity of proteasome subunit low molecular weight protein 2 (LMP2) on cerebral ischemia reperfusion related protein structure of rat blood brain barrier occluden (Occludin) and zonula occludens protein (ZO~ (-1)) on the expression of reperfusion of portal obstruction. Methods MCAO SD in the rat brain (MCAO) model, 1 h ischemia and 72 h. reperfusion in accordance with the weight of the SD rats were randomly divided into 3 groups: sham operation group, experimental group and control group, 10 rats in each group. In MCAO 1 h before operation, the experimental group and the control group received stereotactic brain injection of lentiviral vector LMP2-shRNA and negative control shRNA liquid. With immunoblotting analysis of Occludin, ZO~ (-1) protein expression by Evans blue (EB) tracer monitoring blood brain barrier damage. The results of Occludin protein and ZO~ protein (-1) of the gray value: sham operation 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 the sham group, the control group had significant difference (P0.001); compared with the control group, the experimental group differences were statistically significant (P0.001). The content of EB in brain tissue of sham operation group, control group and experimental group respectively (3.32 + 0.14), (11.41 + 0.21), (6.23 + 0.28) g - g~ (-1), compared with the sham group, control group (P0.001); compared with the control group, there were significant differences between the experimental group (P0.001). Conclusion LMP2 silencing can significantly improve blood after cerebral ischemia in rats brain barrier permeability.

【作者单位】：福建医科大学省立临床医学院福建省立医院神经内科;福建医科大学基础医学院生物化学与分子生物学系;
【基金】：国家自然科学基金面上基金资助项目(81771250) 福建省自然科学基金面上基金资助项目(2016J01432) 福建省卫生系统中青年骨干人才培养项目基金资助项目(2015-ZQN-JC-5)
【分类号】：R743.3

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