YANG Guoqiang,HUANG Jiacheng,YUAN Junjie,et al.Expression and role of peroxiredoxin 1 in rats after intracerebral hemorrhage[J].J Third Mil Med Univ,2020,42(03):300-306.

过氧化物还原酶1在脑出血后的表达变化及作用(/HTML )




Expression and role of peroxiredoxin 1 in rats after intracerebral hemorrhage
YANG Guoqiang HUANG Jiacheng YUAN Junjie ZHANG Qin GONG Changxiong XIE Lexing XIONG Xiaoyi YANG Qingwu
Department of Neurology, Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400037, China
peroxiredoxin 1 intracerebral hemorrhage RNA sequencing
R345; R363.21; R743.34

目的探讨过氧化物还原酶1(peroxiredoxin 1,Prdx1)在中枢神经系统中的细胞定位,以及脑出血(intracerebral hemorrhage,ICH)后Prdx1的表达变化和可能的作用机制。方法取5只8~10周龄SD大鼠脑组织灌注,冰冻切片,免疫荧光染色比较Prdx1在神经元、星形胶质细胞和小胶质细胞中的表达情况。35只8~10周龄SD大鼠按随机数字表法分为sham组、ICH 12 h组、ICH 1 d 组、ICH 2 d 组、ICH 3 d组、ICH 4 d组、ICH 5 d组(n=5),后5组采用自体血注射法建立ICH模型,sham组予以等量生理盐水作为对照。取各组大鼠脑组织,Western blot 和qRT-PCR检测各组中Prdx1 mRNA和蛋白的表达。在HeLa细胞中转染Prdx1干扰质粒或空质粒载体,分为干扰组和对照组,并对两组HeLa细胞进行转录组测序(RNA sequencing,RNA-seq),对差异表达基因(differentially expressed genes,DEGs)进行GO分析,预测Prdx1在ICH中可能的作用。结果免疫荧光染色结果提示:在皮层,Prdx1主要分布于NeuN+的神经元,而在纹状体中,Prdx1主要分布于GFAP+的星形胶质细胞中。Western blot及qRT-PCR结果提示:Prdx1表达在ICH后3 d显著增高(P<0.05),随着时间延长,逐渐降低。RNA-seq及GO分析提示Prdx1引起的差异表达基因主要涉及在炎症反应、细胞凋亡、固有免疫反应、轴突发育等与ICH继发损伤相关的通路。结论Prdx1在不同脑区有不同的细胞分布特点;ICH后,Prdx1升高明显,其在ICH中的可能作用机制与炎症、免疫反应、轴突发育等相关。


Objective To investigate the cellular localization of peroxiredoxin 1 (Prdx1) in the central nervous system, and its expression changes and possible mechanisms after intracerebral hemorrhage (ICH). MethodsFive 8~10-week old SD rats underwent cerebral perfusion, and the brain tissue was sectioned for immunofluorescence staining to observe the localization of Prdx1 in the neurons, astrocytes and microglia. A total of 35 SD rats were randomly divided into sham group, and ICH for 12 h, and 1, 2, 3, 4 and 5 d groups (n=5). The ICH model was established by autologous blood injection in the latter 5 groups, and the sham group was treated with the same amount of normal saline as the control. The expression of Prdx1 at mRNA and protein levels in each group was detected by qRT-PCR and Western blotting. Prdx1 interference plasmid or blank vector were transfected into HeLa cells, and RNA-seq was performed to detect the differentially expressed genes (DEGs) in 2 groups of cells. GO analysis was performed on DEGs to predict the possible role of Prdx1 in ICH. ResultsImmunofluorescence staining showed that Prdx1 was mainly distributed in the NeuN+ neurons in the cortex, while in the striatum, Prdx1 was mainly in the astrocytes. Western blot and qRT-PCR results showed that Prdx1 significantly increased on the 3rd day after ICH (P<0.05), and gradually decreased with elapse of time. RNA-seq and Go analysis suggested that the DEGs caused by Prdx1 were mainly involved in the inflammatory response, apoptosis, innate immune response, axon development and other pathways associated with secondary injury of ICH. ConclusionPrdx1 has different distribution characteristics in different cerebral regions. After ICH, Prdx1 is significantly elevated, and it may be associated with inflammation, immune response, and axon development in ICH. 


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更新日期/Last Update: 2020-02-06