表皮葡萄球菌生物膜抑制肽对该菌黏附及生物膜形成的影响  

Effects of inhibitory peptide of Staphylococcus epidermidis biofilm on adhesion and biofilm formation of this bacterium

作　　者：欧阳净[1] 熊丽蓉[1] 冯伟[1] 孙凤军[1] 陈勇川[1] 

机构地区：[1]第三军医大学西南医院药剂科,重庆400038

出　　处：《中华烧伤杂志》2015年第4期285-289,共5页Chinese Journal of Burns

基　　金：重庆市应用开发计划（cstc2013yykfA10012）

摘　　要：目的研究表皮葡萄球菌生物膜抑制肽（简称抑制肽）对表皮葡萄球菌早期黏附及生物膜形成的影响。方法采用多肽合成仪合成抑制肽，其纯度为96．8％、相对分子质量为874．4。（1）采用终浓度为1～256μg／mL的抑制肽培养表皮葡萄球菌ATCC35984（下同）菌液，不含菌液的M—H肉汤为空白对照，观察抑制肽对该菌的MIC（样本数为3）。（2）采用含终浓度为16、32、64、128、256μg／mL抑制肽的胰蛋白胨大豆肉汤（TSB）培养液培养表皮葡萄球菌菌液（设为相应浓度抑制肽组），以TSB培养基培养前述菌液作为阴性对照组，从培养即刻起每小时观察该菌的生长情况（结果以吸光度值表示），绘制该菌培养24h内的生长曲线（每组各时相点样本数为3）。（3）采用含终浓度为16、32、64、128、256Ixg／mL抑制肽的TSB培养液培养表皮葡萄球菌菌液（设为相应浓度抑制肽组），以TSB培养基培养前述菌液作为阴性对照组，培养4h检测该菌黏附情况，培养20h检测该菌生物膜的形成情况（结果均以吸光度值表示，样本数均为10）。（4）采用含128μg／mL抑制肽的TSB培养液培养表皮葡萄球菌菌液（设为128μg／mL抑制肽组），以TSB培养基培养前述菌液作为阴性对照组，培养24h后采用激光扫描共聚焦显微镜观察该菌的黏附与生物膜形成情况（样本数为3）。对数据行单因素方差分析、LSD检验、DunnettT3检验。结果（1）抑制肽对表皮葡萄球菌的MIC大于256μg／mL。（2）与阴性对照组比较，各浓度抑制肽组表皮葡萄球菌培养24h内的生长曲线无明显变化。（3）256、128、64、32μg／mL抑制肽组培养4h表皮葡萄球菌的黏附情况分别为0．20±0．04、0．27±0．03、0．35．±0．04、0．40．±0．04，明显少于阴性对照组（0．53±0．10，P〈0．05或P〈0．01）；16Ixg／mL抑制肽组培养4h该菌的黏附情况（0．47±0．Objective To study the effects of inhibitory peptide of Staphylococcus epidermidis （SE） biofilm （briefly referred to as inhibitory peptide） on adhesion and biofilm formation of SE at early stage. Methods By using peptide synthesizer, the inhibitory peptide was synthesized with purity of 96.8% and relative molecular mass of 874.4. （ 1 ） Solution of SE ATCC 35984 （ the same below） was cultivated with inhibitory peptide in the final concentrations of 1 - 256 μg/mL, and the M-H broth without bacteria solution was used as blank control. The MIC of the inhibitory peptide against SE was determined （ n = 3）. （2） Solution of SE was cultivated with trypticase soy broth （TSB） culture solution containing inhibitory peptide in the final concentrations of 16, 32, 64, 128, and 256 μg/mL （ set as inhibitory peptide groups in corresponding concentration） , and solution of SE being cultivated with TSB culture medium was used as negative control group. Growth of SE was observed every one hour from immediately after cultivation （denoted as absorbance value）, and the growth curve of SE during the 24 hours of cultivation was drawn, with 3 samples in each group at each time point. （3） Solution of SE was cultivated with TSB culture solution containing inhibitory peptide in the final concentrations of 16, 32, 64, 128, and 256μg/mL （set as inhibitory pcptide groups in corresponding concentration）, and solution of SE being cultivated with TSB culture medium was used as negative control group. Adhesive property of SE was observed after cultivation for 4 hours （ denoted as absorbance value, n = 10） ; biofilm formation of SE was observed after cultivation for 20 hours （ denoted as absorbance value, n = 10）. （4） Solution of SE was cultivated with TSB culture solution containing inhibitory peptide in the final concentration of 128 μg/mL （ set as 128μg/mL inhibitory peptide group） , and solution of SE being cultivated with TSB culture medium was used as negative control group. Adhe

关 键 词：葡萄球菌 表皮 生物膜 抑制肽 

分 类 号：R96[医药卫生—药理学]