檀香醇对金黄色葡萄球菌CS株抑菌活性和机制的研究  

Study on antibacterial activity and the mechanism of santalol against Staphylococcus aureus CS strain

作　　者：邵萌萌 刘鑫[1] 王宇凤 许文博 李升 黄复深[1,2] SHAO Mengmeng;LIU Xin;WANG Yufeng;XU Wenbo;LI Sheng;HUANG Fushen(College of Veterinary Medicine,Hunan Agricultural University,Changsha 410128,China;Hunan Provincial Engineering Technology Research Center for Veterinary Drugs,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]湖南农业大学动物医学院,长沙410128 [2]湖南农业大学湖南兽药工程技术研究中心,长沙410128

出　　处：《黑龙江畜牧兽医》2023年第23期98-102,121,共6页Heilongjiang Animal Science And veterinary Medicine

基　　金：湖南省自然科学基金项目(2017JJ2116)。

摘　　要：为了探讨檀香醇对金黄色葡萄球菌(Staphylococcus aureus,Sa)的抑菌效果和机制,试验以Sa CS株为研究对象,采用倍比稀释法测定檀香醇对Sa CS株的50%最低抑菌浓度(50%minimum inhibitory concentration,MIC50);采用寇氏法计算Sa CS株对小鼠的半数致死量;将40只4周龄的昆明小鼠随机分为4组,先用半数致死量的细菌滴鼻攻毒,各组小鼠每只腹腔注射32,64,128μg/mL的檀香醇0.2 mL,以未经檀香醇处理的小鼠作为对照,正常饲喂4 d,观察小鼠存活情况,试验结束时处死全部小鼠,取肺部组织进行细菌计数,测定檀香醇的体内抑菌效果。以未经檀香醇处理的细菌作为对照,利用SDS-PAGE检测16,32,64,128μg/mL檀香醇对Sa CS株可溶性蛋白表达的影响;采用实时荧光定量PCR方法检测32μg/mL檀香醇对Sa CS株细胞分裂基因和核糖体基因表达的影响。结果表明:檀香醇对Sa CS株的MIC50为32μg/mL。对照小鼠4 d内全部死亡,经32,64,128μg/mL檀香醇处理的小鼠分别存活5,7,9只;檀香醇处理小鼠的肺部细菌数显著或极显著低于对照小鼠(P<0.05或P<0.01)。与对照相比,经64,128μg/mL檀香醇处理6 h后菌体的可溶性蛋白表达量降低。与对照相比,经檀香醇处理的Sa CS株细胞分裂基因ftsZ相对表达量极显著降低(P<0.01);核糖体基因rpsJ相对表达量显著下调(P<0.05),rplB和rpoE基因相对表达量极显著下调(P<0.01),rpsO、rpsA、rpsS和rpsI基因相对表达量极显著上调(P<0.01),rpsC、rpmJ和rplD基因相对表达量差异不显著(P>0.05)。说明檀香醇对Sa CS株具有体内体外抑菌活性,其抑菌机制可能是通过降低Sa CS株的细胞分裂基因、扰乱核糖体基因的表达影响蛋白质表达实现的。In order to investigate the antibacterial effects and the mechanism of santalol against Staphylococcus aureus(Sa),in this experiment,Sa CS strain was used as the research object and the minimum inhibitory concentration(MIC50)of santalol against Sa CS strain was determined by double dilution method.The Karber method was used to calculate half of the lethal dose of Sa CS strain to mice.Forty 4-week-old Kunming mice were randomly divided into 4 groups.First,half of the lethal dose of bacteria were used for challenge by nasal inhalation,and each group of mice was injected with 32,64,128μg/mL of santalol 0.2 mL intraperitoneally;mice untreated with santalol were the control.Normal feeding lasted for 4 days,and the survival of mice was observed;at the end of the experiment,all mice were sacrificed,and lung tissue was taken for bacterial counting,and the antibacterial effect of santalol was measured.SDS-PAGE was used to determine the effect of 16,32,64,128μg/mL santalol on the expression of soluble protein in Sa CS strain,and bacteria without santalol treatment were used as control.The effect of 32μg/mL santalol on the expression of cell division-related genes and ribosomal constituent protein-related genes in Sa CS strain was detected by real-time PCR,and the bacteria treated without santalol were used as control.The results showed that the MIC50 of santalol for Sa CS strain was 32μg/mL.All control mice died within 4 d;5,7,and 9 mice treated with 32,64,128μg/mL santalol survived,respectively.Santalol-treated mice had significantly or very significantly lower lung bacterial counts than the controlled mice(P<0.05 or P<0.01).Compared with the control group,the expression of soluble protein decreased after 6 h of treatment with 64,128μg/mL santalol.Compared with the control group,the relative expression of ftsZ in cell division-related genes in Sa CS strain treated with santalol was significantly reduced(P<0.01).The relative expression of genes rpsJ was significantly downregulated(P<0.05).The relative expression of r

关 键 词：金黄色葡萄球菌CS株 檀香醇 抑菌活性 基因 抑菌机制 

分 类 号：S852.611[农业科学—基础兽医学] S859.7[农业科学—兽医学]