溶血磷脂酶C对铜绿假单胞菌感染小鼠肺功能的作用机制研究
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摘要
目的研究溶血磷脂酶C(plcH)对铜绿假单胞菌肺部感染者肺功能的作用机制,为铜绿假单胞菌肺部感染的靶向治疗提供参考。方法选择8~12周龄成年雄性C57BI/6J小鼠共9只,随机分为3组,即PAO1组、ΔplcH组和对照组(即PBS组),每组各3只,对9只小鼠进行肺感染造模;分别在LB培养基中培养铜绿假单胞菌株PAO1(野生型,即WT型)和等基因ΔplcH缺失株用于小鼠肺感染造模;测定小鼠通气与肺功能,分析小鼠肺泡表面活性物质功能;采用SPSS 19.0软件进行数据统计分析。结果小鼠感染24h后,PAO1组和ΔplcH组的脾脏和肺脏内的铜绿假单胞菌含量差异均无统计学意义;PBS组BALF中的白细胞计数显著低于其他两组(P<0.05);P-V曲线和下降的滞后吸气肢评估结果显示,PAO1组出现极差的肺病理生理学,与PBS组小鼠相比,ΔplcH组小鼠的肺部生理功能显著异常,但好于PAO1组。结论铜绿假单胞菌plcH可能通过作用于肺表面活性物质从而影响肺功能。
OBJECTIVE To study the mechanism of action of hemolytic phospholipase C on pulmonary function of patients with Pseudomonas aeruginosa pulmonary infection so as to provide guidance for targeted treatment of the P.aeruginosa pulmonary infection.METHODS A total of 9C57BI/6Jmale mice aged from 8to 12 weeks were enrolled in the study and randomly divided into three groups:the PAO1 group,theΔplcH group,and the control gorup(the PBS group),with 3mice in each group.The 9mice were used to establish the pulmonary infection models;the P.aeruginosastrains PA01(wild type,known as type WT)and the strains with deletion of allele geneΔplcH were respectively cultured in LB medium so as to establish the mice pulmonary infection models.The ventilation and lung function of the mice were measured,the functions of the pulmonary surfactants were observed,and the statistical analysis of data was performed with the use of SPSS 19.0software.RESULTS There was significant difference in the colony counts of P.aeruginosain spleens or lungs between the PA01 group and theΔplcH group after the mice were infected for 24 hours.The white blood cell counts in BALF were significantly less in the PBS group than in other two groups(P<0.05).The P-V curve and falling delayed inspiratory limb assessment results showed that the PAO1 group had very poor pathophysiology of lungs,as compared with the PBS group,the pulmonary function of theΔplcH group was remarkably abnormal,but it was better than that of the PAO1 group.CONCLUSIONThe P.aeruginosa plcH may affect the lung function through influencing the pulmonary surfactants.
OBJECTIVE To study the mechanism of action of hemolytic phospholipase C on pulmonary function of patients with Pseudomonas aeruginosa pulmonary infection so as to provide guidance for targeted treatment of the P.aeruginosa pulmonary infection.METHODS A total of 9C57BI/6Jmale mice aged from 8to 12 weeks were enrolled in the study and randomly divided into three groups:the PAO1 group,theΔplcH group,and the control gorup(the PBS group),with 3mice in each group.The 9mice were used to establish the pulmonary infection models;the P.aeruginosastrains PA01(wild type,known as type WT)and the strains with deletion of allele geneΔplcH were respectively cultured in LB medium so as to establish the mice pulmonary infection models.The ventilation and lung function of the mice were measured,the functions of the pulmonary surfactants were observed,and the statistical analysis of data was performed with the use of SPSS 19.0software.RESULTS There was significant difference in the colony counts of P.aeruginosain spleens or lungs between the PA01 group and theΔplcH group after the mice were infected for 24 hours.The white blood cell counts in BALF were significantly less in the PBS group than in other two groups(P<0.05).The P-V curve and falling delayed inspiratory limb assessment results showed that the PAO1 group had very poor pathophysiology of lungs,as compared with the PBS group,the pulmonary function of theΔplcH group was remarkably abnormal,but it was better than that of the PAO1 group.CONCLUSIONThe P.aeruginosa plcH may affect the lung function through influencing the pulmonary surfactants.
引文
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[2]Lee J,Zhang L.The hierarchy quorum sensing network in Pseudomonas aeruginosa[J].Protein Cell,2015,6(1):26-41.
[3]Imperi F,Leoni L,Visca P.Antivirulence activity of azithromycin in Pseudomonas aeruginosa[J].Front Microbiol,2014,(5):178.
[4]Lopez DJ,Collado MI,Ibarguren M,et al.Multiple phospholipid substrates of phospholipase c/sphingomyelinase HR2from Pseudomonas aeruginosa[J].Chem Phys Lipids,2011,164(1):78-82.
[5]Massa CB,Scott P,Abramova E,et al.Acute chlorine gas exposure produces transient inflammation and a progressive alteration in surfactant composition with accompanying mechanical dysfunction[J].Toxicol Appl Pharmacol,2014,278(1):53-64.
[6]刘晓荣,翟俊伟,张晓群,等.医院ICU患者下呼吸道绿脓杆菌感染的耐药性分析[J].临床合理用药杂志,2014,7(10):42-43.
[7]余瑛,郭冰峰,郭志龙,等.N端色氨酸修饰增强hemokinin-1拮抗绿脓杆菌的活性[J].重庆理工大学学报:自然科学版,2014,28(12):55-58.
[8]Wargo MJ.Choline catabolism to glycine betaine contributes to Pseudomonas aeruginosasurvival during murine lung infection[J].PLoS One,2013,8(2):e56850.
[9]Beaufort N,Corvazier E,Mlanaoindrou S,et al.Disruption of the endothelial barrier by proteases from the bacterial pathogen Pseudomonas aeruginosa:implication of matrilysis and receptor cleavage[J].PLoS One,2013,8(9):e75708.
[10]Kuang Z,Hao Y,Hwang S,et al.The Pseudomonas aeruginosa flagellum confers resistance to pulmonary surfactant protein-a by impacting the production of exoproteases through quorum-sensing[J].Mol Microbiol,2011,79(5):1220-1235.