三叶青提取物TH-w3对金黄色葡萄球菌生物膜的抑制和清除作用  

Inhibitory and scavenging effects of cloverleaf extract TH-w3 on Staphylococcus aureus biofilm

作　　者：孙丹 姬会春 祝宇翀 单宇 刘军权 廖雨琴 Sun Dan;Ji Huichun;Zhu Yuchong;Shan Yu;Liu Junquan;Liao Yuqin(Department of Clinical Laboratory,The First People's Hospital of Linping District,Hangzhou Zhejiang 311100,China;Department of Infectious Diseases,Suqian First People's Hospital,Suqian Jiangsu 223800,China;Hangzhou KingMed Medical Laboratory Co.,Ltd.,Hangzhou Zhejiang 310053,China)

机构地区：[1]临平区第一人民医院检验科,浙江杭州311100 [2]宿迁市第一人民医院传染病科,江苏宿迁223800 [3]杭州金域医学检验所有限公司,浙江杭州310053

出　　处：《中华临床实验室管理电子杂志》2022年第4期227-232,共6页Chinese Journal of Clinical Laboratory Management(Electronic Edition）

摘　　要：目的探讨三叶青提取物TH-w3对金黄色葡萄球菌生物膜形成的抑制和清除作用。方法TH-w3是用80%的乙醇提取后的三叶青干燥块根水煮后减压回收蒸干产品;实验菌株为本实验室临床分离培养并保存的耐甲氧西林金黄色葡萄球菌(MRSA)和甲氧西林敏感金黄色葡萄球菌(MSSA)各30株;菌落计数法绘制金黄色葡萄球菌生物膜形成曲线;二倍稀释法肉眼观察+酶标仪比浊法测定最小抑菌浓度(MIC);结晶紫法检测TH-w3对金黄色葡萄球菌生物膜的抑制及清除作用。结果对各30株MRSA和MSSA的成膜能力进行比较,MRSA菌株成膜阳性22株(73.3%),MSSA成膜阳性菌株3株(10.0%),成膜阳性率MRSA菌株与MSSA菌株比较差异有统计学意义(χ^(2)=43.35,P<0.01);TH-w3对“MRSA(+)组”和“MSSA(+)组”菌株的MIC均为8.00μg/ml;TH-w3对“MRSA(+)组”和“MSSA(+)组”菌株生物膜的抑制和清除作用均具有浓度依赖性,在1个MIC浓度下抑制率和清除率最高,可达68%以上。相同MIC的TH-w3对金黄色葡萄球菌生物膜的抑制和清除作用随培养时间延长明显升高,至48 h后提升渐缓。在1/4 MIC浓度下,诱导24 h后MSSA(+)的抑制率高于MRSA(+)(P<0.05)。在1/4和1/2 MIC浓度下,诱导24 h后MSSA(+)的清除率高于MRSA(+)(P<0.01)。结论三叶青提取物TH-w3对金黄色葡萄球菌生物膜有明显抑制和清除作用,具有临床转化价值。Objective To explore the inhibition and scavenging effects of TH-w3 extracted from Tetrastigma hemsleyanum on the formation of Staphylococcus aureus biofilm.Methods TH-w3 was prepared from dried root of Tetrastigma hemsleyanum by reducing pressure and boiling dry after extraction with 80%ethanol.30 strains of Methicillin-resistant Staphylococcus aureus(MRSA)and methicillin-sensitive staphylococcus aureus(MSSA)were isolated,cultured and preserved respectively.Colony-counting method was used to draw the biofilm formation curve of Staphylococcus aureus.The minimum inhibitory concentration(MIC)of TH-w3 were determined by visual inspection and absorbance detection via double ratio dilution method.The inhibition and scavenging effects of TH-w3 on the formation of Staphylococcus aureus biofilm were observed by Crystal violet staining.Results 22 strains of MRSA(73.3%)and 3 strains of MSSA(10.0%)were observed positive membrane formation.The biofilm formation rates between MRSA and MSSA were statistically significant(χ^(2)=43.35,P<0.01).The MIC of TH-w3 for MRSA and MSSA in positive biofilm-forming groups were both 8.00μg/ml.The inhibition and scavenging effects of TH-w3 on MRSA and MSSA showed concentration-dependent,and the inhibition and scavenging rates reached the highest(more than 68%)at the concentration of 1 MIC.The inhibitory and clearing effects of TH-w3 with the same MIC on the biofilm of Staphylococcus aureus expressed a smooth increase after 48 hrs.The inhibition and scavenging rates for MSSA showed higher than MRSA in 1/2 and 1/4 MIC group after induction for 24 hours(P<0.05).Conclusions The trifoliate extract TH-w3 showed significant inhibition and scavenging effects on Staphylococcus aureus biofilm,which promised a potential clinical translational value.

关 键 词：三叶青 金黄色葡萄球菌 生物膜 抑制作用 清除作用 

分 类 号：R446.5[医药卫生—诊断学]