川乌水提取物及其单体成分对金黄色葡萄球菌α-溶血素分泌的抑制作用  

Inhibitory Effect of Aconiti Radix Water Extract and Its Monomeric Components on the Secretion ofα-Hemolysin in Staphylococcus Aureus

作　　者：李詹 谈玲 丁曼琳 黄远帅 LI Zhan;TAN Ling;DING Manlin;HUANG Yuanshuai(Southwest Medical University Affiliated Hospital,Luzhou,Sichuan,China 646000)

机构地区：[1]西南医科大学附属医院,四川泸州646000

出　　处：《中国药业》2024年第13期51-55,共5页China Pharmaceuticals

基　　金：国家自然科学基金面上项目[81974299]。

摘　　要：目的探讨川乌水提取物及其单体成分对金黄色葡萄球菌α-溶血素分泌的抑制作用。方法采用转录组测序技术(RNAseq)分析川乌水提取物对金黄色葡萄球菌USA300 hla,psma,isda基因转录水平的影响。采用反转录-实时荧光定量聚合酶链式反应(RT-qPCR)法测定川乌水提取物及质量浓度分别为0.01,0.1,1 mg/mL的乌头原碱、次乌头碱、新乌头碱单体成分对hla,psma,isda基因表达水平的影响,采用溶血定量试验考察溶血活性。采用免疫印迹(Western blot)法考察质量浓度分别为0.01,0.1,1 mg/mL的乌头原碱、次乌头碱、新乌头碱单体成分对金黄色葡萄球菌菌液上清液中α-溶血素分泌的影响。结果USA300经250 mg/mL川乌水提取物处理后,α-溶血素、δ-溶血素、酚溶性调节蛋白等毒力因子相关基因的表达均显著下调(P<0.005)。USA300经250 mg/mL川乌水提取物及1 mg/mL乌头原碱、次乌头碱、新乌头碱单体成分处理后,hla,psma,isda基因转录水平均受到显著抑制(P<0.05),其中对hla基因的抑制作用最显著;金黄色葡萄球菌的溶血活性均受到显著抑制(P<0.01)。USA300经0.01,0.1,1 mg/mL乌头原碱、次乌头碱、新乌头碱单体成分处理后,金黄色葡萄球菌菌液上清液中α-溶血素分泌量与乌头原碱呈浓度依赖性降低,但经次乌头碱、新乌头碱单体成分处理后的趋势不明显。结论川乌水提取物及其单体成分可抑制金黄色葡萄球菌中α-溶血素的分泌,尤其是单体成分乌头原碱有用于抗金黄色葡萄球菌感染的潜能。Objective To investigate the inhibitory effect of Aconiti Radix water extract and its monomeric components on the secretion ofα-hemolysin in Staphylococcus aureus.Methods RNAseq was used to analyze the effect of Aconiti Radix water extract on the transcription levels of USA300 hla,psma,and isda genes in Staphylococcus aureus.The reverse transcription real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)method was used to determine the effect of Aconiti Radix water extract and its monomeric components of aconitine(0.01 mg/mL),hypaconitine(0.1 mg/mL),and mesaconitine(1 mg/mL)on the expression levels of hla,psma,and isda genes.The quantitative hemolysis test was used to evaluate the hemolytic activity.Western blot was used to investigate the effect of monomeric components of aconitine(0.01 mg/mL),hypaconitine(0.1 mg/mL),and mesaconitine(1 mg/mL)on the secretion ofα-hemolysin in the supernatant of Staphylococcus aureus bacterial solution.Results After USA300 was treated with Aconiti Radix water extract(250 mg/mL),genes related to virulence factors such asα-hemolysin,δ-hemolysin,and phenol-soluble regulatory proteins were significantly down-regulated(P<0.005).After USA300 was treated with the Aconiti Radix water extract(250 mg/mL)and its monomeric components of aconitine(1 mg/mL),hypaconitine(1 mg/mL),and mesaconitine(1 mg/mL),the transcript levels of hla,psma,and isda genes were significantly inhibited(P<0.05),with the most significant inhibition level of the hla gene.The hemolytic activity of Staphylococcus aureus was significantly inhibited(P<0.01).After USA300 was treated with the monomeric components of aconitine(0.01 mg/mL),hypaconitine(0.1 mg/mL),and mesaconitine(1 mg/mL),the secretion ofα-hemolysin in the supernatant of Staphylococcus aureus decreased in a concentration dependent manner with aconitine,but the trend was not significant after treatment with monomeric components of hypaconitine and mesaconitine.Conclusion The Aconiti Radix water extract and its monomeric components can inhib

关 键 词：金黄色葡萄球菌 川乌 α-溶血素 乌头原碱 次乌头碱 新乌头碱 

分 类 号：R932[医药卫生—生药学] R285.5[医药卫生—药学]