目的 以秀丽隐杆线虫铜绿假单胞菌(pseudomonas aeruginosa,PA)感染模型,评价美罗培南的体内抗菌作用及机制.方法 PA感染秀丽隐杆线虫,用培养液配制不同浓度的药物进行有效药物作用评价和筛选;将线虫置入药液培养,以秀丽线虫感染致死率判断药效;蛋白印迹法检测秀丽线虫感染后促分裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)活性的改变及美罗培南作用.PCR方法检测秀丽线虫抗菌肽基因Lys-1,clec-85,F55G11.7,K08D8.5的表达活性变化及美罗培南对其的影响.结果与对照组比较,PA感染模型组的秀丽线虫在致死率方面发生明显变化,抗菌药物美罗培南对感染细菌的秀丽隐杆线虫显示出较好的保护效果(P<0.01).对MAPK激酶活性表达的检测表明,PA感染可致PMK-1激酶活性升高.但同时发现,对照组OP-50大肠杆菌饲养组线虫的PMK-1激酶也被激活,而美罗培南并不影响PA感染组秀丽线虫的PMK-1激酶活性.在PA感染模型组,秀丽线虫的抗菌肽基因Lys-1,clec-85,F55G11.7,K08D8.5的表达活性升高(P<0.01);而抗菌药物美罗培南进一步促进这些抗菌肽基因的表达增高(P<0.01),表现出一定的协同作用.结论 通过PA培养可以建立稳定的秀丽线虫感染模型,此模型可应用于病原菌感染敏感药物快速简便筛选.
Effect of meropenem on killing of caenorhabditis elegans by pseudomonas aeruginosa
Objective To evaluate antimicrobial effect and mechanism of meropenem in the model of PA infection by C.elegans.Methods To evaluate drug effects of PA infection with caenorhabditis elegans by different concentrations of culture medium, determinate the lethal rate of C.elegans.Western blot detected mitogen activated protein kinase ( Mitogen-activated protein kinase MAPK ) activity change, and PCR detected antimicrobial peptide genes expression in C.elegans after PA infection,the effect of meropenem on MAPK activity change and antimicrobial peptide genes expression.Results Compared with the control group (OP-50), the death rate of C.elegans in PA infection group changed significantly (P<0.01). Meropenem showed protective effect after C.elegans infection ( P <0.01 ) .Detection of MAPK kinase activity showed that PA infection caused PMK-1 kinase activation, further study showed that antibiotics meropenem did not affect the activation of PMK-1 kinase (no significant difference).C.elegans antimicrobial peptide gene Lys-1, clec-85, F55G11.7, K08D8.5 activity increased in PA infection (P<0.01).Meropenem promoted the expression of the antimicrobial peptide gene increased (P<0.01),with synergistic effects.Conclusion Our results show that a C.elegans pathogenicity model can be applied screening drug susceptible to pathogens infection quickly and easily.
NETL
NSTL
万方数据
维普
