Effect of inhibition of nitric oxide synthase on Pseudomonas aeruginosa infection of respiratory mucosa in vitro
Effect of inhibition of nitric oxide synthase on Pseudomonas aeruginosa infection of respiratory mucosa in vitro
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DOI：
10.1165/ajrcmb.19.6.2904
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发表时间：
1998-12-01
影响因子：
6.4
通讯作者：
Wilson, R
中科院分区：
文献类型：
--
作者：
Dowling, RB;Newton, R;Wilson, R
我们研究了一氧化氮合酶（NOS）抑制剂不对称二甲基精氨酸（ADMA）和非活性对映体N-G-甲基-D-精氨酸（D-NMMA）对铜绿假单胞菌感染的呼吸道粘膜在鼻甲器官培养的影响。我们还研究了铜绿假单胞菌培养滤液对上皮细胞系（A549）诱导型NOS（iNOS）信使RNA（mRNA）表达的影响。在细菌感染前，将器官培养物与ADMA（0.1至4 x 10（-4）M）或D-NMMA（2 x 10（-4）M）预孵育30 min。感染的器官培养物（8小时）有显着（P小于或等于0.05）更大的上皮细胞损伤和更少的纤毛和unciliated细胞比对照培养。与对照培养物相比，在喂养感染器官培养物的培养基中亚硝酸盐水平增加。ADMA以浓度依赖性方式显著（P <或等于0.05）减少细菌诱导的上皮损伤和纤毛细胞损失。D-NMMA不影响铜绿假单胞菌感染粘膜的效果。ADMA，但不是D-NMMA，显着（P小于或等于0.04）减少粘附到呼吸道粘膜的细菌总数。铜绿假单胞菌培养24 h和36 h显著（P = 0.02）增加iNOS的甘油醛-3-磷酸脱氢酶mRNA表达。这些结果表明，铜绿假单胞菌刺激细胞系的iNOS表达和器官培养物的NO产生。ADMA。减少粘膜损伤和纤毛细胞的损失，这表明NO可能是铜绿假单胞菌引起的上皮损伤的介质。
We studied the effect of the nitric oxide synthase (NOS) inhibitor asymmetric dimethyl arginine (ADMA) and the inactive enantiomer N-G-methyl-D-arginine (D-NMMA) on Pseudomonas aeruginosa infection of the respiratory mucosa in nasal turbinate organ cultures. We also investigated the effect of P. aeruginosa culture filtrate on the expression of inducible NOS (iNOS) messenger RNA (mRNA) by an epithelial cell line (A549). Organ cultures were preincubated with ADMA (0.1 to 4 x 10(-4) M) or D-NMMA (2 x 10(-4) M) for 30 min prior to bacterial infection. Infected organ cultures (8 h) had significantly (P less than or equal to 0.05) greater epithelial damage and fewer ciliated and unciliated cells than did control cultures. There was an increased level of nitrite in the medium feeding infected organ cultures as compared with control cultures. ADMA significantly (P less than or equal to 0.05) reduced both bacterially induced epithelial damage and loss of ciliated cells in a concentration-dependent manner. D-NMMA did not influence the effect of P. aeruginosa infection of the mucosa. ADMA, but not D-NMMA, significantly (P less than or equal to 0.04) reduced total bacterial numbers adherent to the respiratory mucosa. P. aeruginosa culture filtrates (24 h and 36 h) significantly (P = 0.02) increased iNOS with respect to glyceraldehyde-3-phosphate dehydrogenase mRNA expression. These results show that P. aeruginosa stimulates iNOS expression by a cell Line and NO production by an organ culture. ADMA. reduces mucosal damage and loss of ciliated cells, which suggests that NO may be a mediator of epithelial damage caused by P. aeruginosa.