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白细胞介素1β刺激对脑微血管内皮细胞COX活性、mRNA表达和PGE_2释放的影响
Alternative TitleStudy on COX activity and its mRNA expression,and PGE_2 release from cerebral microvascular endothelial cells stimulated by IL-1β
郭建友; 霍海如; 赵保胜; 刘洪斌; 李兰芳
2007
Source Publication中国病理生理杂志
ISSN1000-4718
Volume23Issue:4Pages:710-714
Abstract目的:探讨白细胞介素1β(IL-1β)刺激不同时间对大鼠脑微血管内皮细胞(rCMEC)的环氧合酶(COX)活性及其mRNA表达和PGE2释放的影响。方法:建立rCMEC培养,进行Ⅷ因子相关抗原鉴定。细胞长至融合状态后加入IL-1β(30μg/L),分别刺激0.5、1、2、4、8、12、24 h,ELISA测定细胞内COX-1、COX-2的活性及细胞外液PGE2含量,荧光实时定量PCR检测COX-1、COX-2的mRNA表达量,扩增产物进行熔解曲线图及琼脂糖电泳分析,并与未刺激的rCMEC作比较。结果:①Ⅷ因子抗体免疫组化染色可见90%以上的培养细胞呈阳性,确认为rCMEC。②IL-1β刺激4 h时细胞培养液PGE2含量已明显高于未刺激组(P<0.05);12 h时PGE2含量达到最大值(P<0.01);24 h时PGE2含量有所回降,但与未刺激组比较仍有显著差异(P<0.05)。③IL-1β刺激不同时间rCMEC内COX-1活性与未刺激组相比无统计学差异(P>0.05);COX-2活性在第8 h时已明显高于未刺激组(P<0.05),12 h活性达峰值(P<0.01),24 h活性有所回降,但仍具显著差异(P<0.05)。④IL-1β刺激不同时间COX-1 mRNA表达与未刺激组比较无明显差异(P>0.05);未刺激组在本实验条件下未检测到COX-2 mRNA表达,IL-1β刺激1h时可见COX-2 mRNA表达,4 h时COX-2 mRNA表达至峰值,而后开始回降,第12 h时已未见表达。熔解曲线图显示无非特异性扩增;琼脂糖电泳可见扩增基因与目的基因长度相符,结果与荧光定量PCR一致。结论:IL-1β作用下,rCMEC释放的PGE2持续增加并于12h达峰值,这一过程主要与COX-2 mRNA表达激活及COX-2活性增加有关。
Keyword白细胞介素1 内皮细胞 前列腺素E类 环氧合酶-2
Subject Area系统生物学/生物信息学
Indexed By中文核心期刊要目总览 ASPT来源刊 CJFD收录刊
Language中文
Funding Project国家自然科学基金重大项目资助(No.90209006)
NoteAIM: To study the cyclooxygenase(COX) activity and its mRNA expression,and PGE2 release from rats cerebral microvascular endothelial cells(rCEMC) stimulated by IL-1β(30 μg/L) at different times.METHODS: rCMEC were cultured,and identified by immunohistochemistry for von Willebrand factor(Ⅷ factor,a marker for all endothelial cells) in cytoplasm of the cells.After rCEMC grew to confluency,they were stimulated with IL-1β for 0.5,1,2,4,8,12 and 24 h,respectively.Activity of COX-1 and COX-2 in rCEMC and producti...
Document Type期刊论文
Identifierhttp://ir.psych.ac.cn/handle/311026/3332
Collection中国科学院心理研究所回溯数据库(1956-2010)
Corresponding Author霍海如
Recommended Citation
GB/T 7714
郭建友,霍海如,赵保胜,等. 白细胞介素1β刺激对脑微血管内皮细胞COX活性、mRNA表达和PGE_2释放的影响[J]. 中国病理生理杂志,2007,23(4):710-714.
APA 郭建友,霍海如,赵保胜,刘洪斌,&李兰芳.(2007).白细胞介素1β刺激对脑微血管内皮细胞COX活性、mRNA表达和PGE_2释放的影响.中国病理生理杂志,23(4),710-714.
MLA 郭建友,et al."白细胞介素1β刺激对脑微血管内皮细胞COX活性、mRNA表达和PGE_2释放的影响".中国病理生理杂志 23.4(2007):710-714.
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