黄芩射干汤对金黄色葡萄球菌泛毒力的实验研究  

Study on pan-virulence of Huangqin Shegan Decoction against Staphylococcus aureus

作　　者：刘畅[1] 宋伍 姜涛[1] 李雨芬 周豪芳 苏鑫[1] LIU Chang;SONG Wu;JIANG Tao;LI Yu-fen;ZHOU Hao-fang;SU Xin(Changchun University of Chinese Medicine,Chang chun 130117 China)

机构地区：[1]长春中医药大学,吉林长春130117

出　　处：《时珍国医国药》2024年第11期2566-2571,共6页Lishizhen Medicine and Materia Medica Research

基　　金：吉林省教育厅科学技术研究项目(JJKH20230985KJ)。

摘　　要：目的研究最低抑菌浓度(MIC)以下浓度的黄芩射干汤对金黄色葡萄球菌(金葡菌)的体外毒力作用的影响。方法首先采用微量稀释法和时间-生长曲线测定黄芩射干汤对金葡菌的MIC和对细菌生长的影响;用溶血实验和蛋白质印迹实验来探究MIC浓度以下黄芩射干汤对金葡菌的溶血作用和α-溶血素(Hla)分泌的影响;用脲酶活性实验来研究黄芩射干汤对金葡菌毒力表达的影响;用纤维蛋白原黏附实验和生物膜形成实验研究MIC浓度以下黄芩射干汤对金葡菌生物膜形成和黏附能力的影响;用流式细胞仪来研究MIC浓度以下黄芩射干汤对金葡菌表面蛋白A(SpA)锚定的影响。结果黄芩射干汤对金葡菌USA300的MIC为16 mg/mL,对Newman的MIC为16 mg/mL,对SA37的MIC为32 mg/mL。在不明显抑菌的情况下,MIC以下浓度的黄芩射干汤表现出对金葡菌USA300溶血能力、生物膜形成和黏附能力的抑制作用,并可以减少Hla的分泌;可以通过抑制SrtA的活性来减少细胞壁中SpA锚定的含量。此外,推测黄芩射干汤可能是通过抑制ClpP的活性而抑制金葡菌毒力表型。结论MIC浓度下的黄芩射干汤在体外减弱了金葡菌毒力相关的表型,给金葡菌的治疗提供中医药选择。Objective To study the effect of Huangqin Shegan Decoction below the minimum inhibitory concentration(MIC)on the virulence of Staphylococcus aureus(S.aureus)in vitro.Methods Microdilution method and time-growth curve were used to determine the MIC of Huangqin Shegan Decoction on S.aureus and the effect on bacterial growth.The effects of Huangqin Shegan Decoction on the hemolysis andα-hemolysin(Hla)secretion of S.aureus under MIC concentration were investigated by hemolysis test and western blot test.The effect of Huangqin Shegan Decoction on the expression of virulence of S.aureus was studied by urease activity experiment.The effects of Huangqin Shegan Decoction under MIC concentration on the biofilm formation and adhesion of S.aureus were studied by fibrinogen adhesion experiment and biofilm formation experiment.The effect of Huangqin shegan Tang under MIC concentration on SpA anchoring of S.aureus was studied by flow cytometry.Results The MIC of Huangqin Shegan Decoction was 16 mg/mL for S.aureus USA300,16 mg/mL for Newman and 32 mg/mL for SA37.Under the condition of no obvious bacteriological inhibition,Huangqin Shegan Decoction with MIC concentration below showed inhibitory effects on the hemolytic ability,biofilm formation and adhesion ability of S.aureus USA300,and could reduce the secretion of Hla.The amount of SpA anchoring in the cell wall can be reduced by inhibiting the activity of SrtA.In addition,it was speculated that Huangqin Shegan Decoction might inhibit the virulence phenotype of S.aureus by inhibiting the activity of ClpP.Conclusion Huangqin Shegan Decoction with MIC concentration can attenuate the virulence related phenotype of S.aureus in vitro and provide a choice of traditional Chinese medicine for the treatment of S.aureus.

关 键 词：黄芩射干汤 金黄色葡萄球菌 溶血 生物膜 毒力 

分 类 号：R285.5[医药卫生—中药学]