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铜绿假单胞菌感染小鼠眼睛中促炎细胞因子和趋化因子的基因表达。

Gene expression of pro-inflammatory cytokines and chemokines in mouse eye infected with Pseudomonas aeruginosa.

作者信息

Xue Mei-Lang, Thakur Archana, Willcox Mark

机构信息

Cooperative Research Centre For Eye Research and Technology, School of Optometry, University of New South Wales, Sydney, New South Wales, Australia.

出版信息

Clin Exp Ophthalmol. 2002 Jun;30(3):196-9. doi: 10.1046/j.1442-9071.2002.00510.x.

Abstract

Ocular infection with Pseudomonas aeruginosa triggers extensive host inflammatory response and corneal damage. The purpose of present study was to investigate the gene expression of pro-inflammatory mediators interleukin (IL)-1 beta, IL-6, tumour necrosis factor-alpha (TNF-alpha),macrophage inflammatory protein (MIP)-2 and cytokine-induced neutrophil chemoattractant (KC) in the mouse eye challenged with P. aeruginosa. Scratched mouse corneas were infected with three phenotypes of P. aeruginosa individually. Total RNA was extracted from mouse eyes at 4 h, 8 h,16 h and 24 h post-challenge. Single stranded cDNA was synthesized from total RNA by reverse transcription and then subjected to polymerase chain reaction (PCR) using specific primers for IL-1 beta, IL-6, TNF-alpha, MIP-2 and KC. Results revealed three patterns of cytokines and chemokines expression in response to ocular infection with three phenotypes of P. aeruginosa. Ocular infection with the invasive strain induced the highest levels of IL-1 beta, IL-6, MIP-2 and KC mRNA, followed by the infection with the cytotoxic strain. Ocular infection with the CLARE strain induced the lowest levels of IL-1 beta, IL-6, MIP-2 and KC mRNA. The expression of TNF-alpha mRNA was very low and irregular following P. aeruginosa challenge. These data indicate that over-expression of pro-inflammatory cytokines and chemokines may represent a vigorous immune response and therefore may contribute to corneal damage during P. aeruginosa infection.

摘要

铜绿假单胞菌引起的眼部感染会引发广泛的宿主炎症反应和角膜损伤。本研究的目的是调查在用铜绿假单胞菌攻击的小鼠眼中促炎介质白细胞介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)、巨噬细胞炎性蛋白(MIP)-2和细胞因子诱导的中性粒细胞趋化因子(KC)的基因表达。分别用三种铜绿假单胞菌表型感染划痕的小鼠角膜。在攻击后4小时、8小时、16小时和24小时从小鼠眼中提取总RNA。通过逆转录从总RNA合成单链cDNA,然后使用针对IL-1β、IL-6、TNF-α、MIP-2和KC的特异性引物进行聚合酶链反应(PCR)。结果揭示了三种细胞因子和趋化因子表达模式,以响应三种铜绿假单胞菌表型的眼部感染。用侵袭性菌株进行眼部感染诱导了最高水平的IL-1β、IL-6、MIP-2和KC mRNA,其次是细胞毒性菌株感染。用CLARE菌株进行眼部感染诱导了最低水平的IL-1β、IL-6、MIP-2和KC mRNA。在铜绿假单胞菌攻击后,TNF-αmRNA的表达非常低且不规则。这些数据表明促炎细胞因子和趋化因子的过度表达可能代表一种强烈的免疫反应,因此可能在铜绿假单胞菌感染期间导致角膜损伤。

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