Effects of RNA interference targeting low molecular weight protein 2 on blood brain barrier after cerebral ischemia in a rat model
10.13699/j.cnki.1001-6821.2017.17.022
- VernacularTitle: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
- Author:
Xing-Yong CHEN
1
;
Xu ZHANG
;
Hui-Xin LEI
;
Zhi-Jun LIAO
;
Yin-Zhou WANG
Author Information
1. 福建医科大学省立临床医学院、福建省立医院神经内科
- Keywords:
immunoproteasome;
RNA interference targeting low molecular weight protein 2;
blood brain barrier;
cerebral ischemia
- From:
The Chinese Journal of Clinical Pharmacology
2017;33(17):1687-1689,1705
- CountryChina
- Language:Chinese
-
Abstract:
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 induced 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 (n =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 Occludin 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 Oceludin 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,th e 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).Conclusion RNA interference targeting silence LMP2 significantly improved the stability of BBB after cerebral ischemia.
