CAJ | 학술논문

目的观察RNA干扰免疫蛋白酶体亚基低分子重量蛋白2 ( LMP2 )对大鼠脑缺血再灌注后神经炎症反应的影响. 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. SD大鼠按照体重随机分为假手术组、实验组和对照组,每组10只. 实验组和对照组分别在MCAO术前1 h,立体定位脑内注射慢病毒载体LMP2-shRNA及阴性对照shRNA液体. 用Western blot法分析LMP2、LMP7、磷酸化核因子-κB p65 ( NF-κB p65 )蛋白表达,用ELISA法测定脑组织白介素-1β( IL-1β)、肿瘤坏死因子-α( TNF-α)的浓度. 结果局灶性脑缺血再灌注后,梗死灶周边的皮层和纹状体区LMP2、NF-κB、IL-1β和TNF-α蛋白表达上调,注射慢病毒LMP2-shRNA载体可显著下调这些蛋白产生,减少脑梗死体积,且未见明显的药物不良反应. 结论RNA干扰LMP2表达,可显著下调脑缺血后神经炎症反应,发挥神经保护作用.
목적 관찰RNA간우면역단백매체아기저분자중량단백2 ( LMP2 )대대서뇌결혈재관주후신경염증반응적영향. 방법 선전법제작SD대서대뇌중동맥조새재관주( MCAO)모형,뇌결혈1 h재관주72 h. SD대서안조체중수궤분위가수술조、실험조화대조조,매조10지. 실험조화대조조분별재MCAO술전1 h,입체정위뇌내주사만병독재체LMP2-shRNA급음성대조shRNA액체. 용Western blot법분석LMP2、LMP7、린산화핵인자-κB p65 ( NF-κB p65 )단백표체,용ELISA법측정뇌조직백개소-1β( IL-1β)、종류배사인자-α( TNF-α)적농도. 결과 국조성뇌결혈재관주후,경사조주변적피층화문상체구LMP2、NF-κB、IL-1β화TNF-α단백표체상조,주사만병독LMP2-shRNA재체가현저하조저사단백산생,감소뇌경사체적,차미견명현적약물불량반응. 결론RNA간우LMP2표체,가현저하조뇌결혈후신경염증반응,발휘신경보호작용.
Objective To investigate the effects of RNA interference targeting immunoproteasome subunit low molecular weight protein 2 ( LMP2 ) on the neuroinflammatory reaction after cerebral ischemia/reperfusion in a rat model .Methods The middle cerebral artery occlu-sion( MCAO) was induced using the intraluminal suture occlusion tech-nique in SD rats.Reperfusion of cerebral blood flow was allowed by gen-tly removing the monofilament after 1 hour ischemia , followed by 72 h reperfusion.Rats were randomly assigned into 3 groups:sham-operated group, experimental group and control group ( n=10).The preparations of lentivirus vector targeting LMP2 short hairpin RNA (shRNA) and ca-rrying scrambled shRNA were infused stereotactically into the ipsilateral hemispheric region 1 h before MCAO in the experimental group and con-trol group, respectively.The expressions of LMP2, phospho -nuclear factor-κB p65 ( NF-κB P-p65 ) protein and levels of interleukin -1β( IL-1β) and tumor necrosis factor -α( TNF-α) protein in the brain were detected by Western blot and enzyme -linked immunosorbent assay (ELISA), respectively.Results The expression of LMP2, NF-κB, IL -1βand TNF -αincreased significantly higher in the cortex and striatum surrounding the infarct core than that in the sham -operation group ( P<0.001 ) .Pretreated with LMP2 -shRNA 1 h before MCAO significantly decreased the levels of LMP2, NF-κB, IL-1βand TNF-αprotein, more importantly.The shRNA-mediated inhibition of LMP2 efficiently reduced infarction volume and without significant adverse drug reaction . Conclusion The shRNA-mediated inhibition of LMP2 significantly reduces neuroinflammatory reaction and displays neuroprotective effect.