藤茶提取物及活性成分对创伤弧菌的体外抑菌活性及作用机制  

In vitro antibacterial activity and mechanism of Ampelopsis grossedentata extract and its active components against Vibrio vulnificus

作　　者：祁子麟 陈咏琪 蔡延渠[2,3,4] 李苑新 鲁湘鄂[2,3,4] 张萱萱 QI Zilin;CHEN Yongqi;CAI Yanqu;LI Yuanxin;LU Xiange;ZHANG Xuanxuan(School of Medical Department,Hubei University of Science and Technology,Xianning 437100,China;Center for Drug Research and Development,Guangdong Pharmacological University,Guangzhou 510006,China;ClassⅢLaboratory of Modern Chinese Medicine Preparation,State Administration of Traditional Chinese Medicine of China,Guangzhou 510006,China;Key Laboratory of Modern Chinese Medicine of Education Department of Guangdong Province,Guangzhou 510006,China;Guang Provincial Key Laboratory of Aduanced Drug Dilivery Systens,Guagnzhou 510006,China)

机构地区：[1]湖北科技学院医学部,湖北咸宁437100 [2]广东药科大学新药研发中心,广东广州510006 [3]国家中医药管理局中药制剂实验室(三级),广东广州510006 [4]广东省教育厅现代中药重点实验室,广东广州510006 [5]广东省药物新剂型重点实验室,广东广州510006

出　　处：《广东药科大学学报》2023年第1期95-99,共5页Journal of Guangdong Pharmaceutical University

基　　金：广东省驻镇帮镇扶村农村科技特派员项目(KTP20210273);广东省医学科研基金项目(A2020368)。

摘　　要：目的 研究藤茶提取物及活性成分对创伤弧菌的体外抑菌活性和作用机制,为其开发应用提供实验依据。方法 通过加热回流提取法制备藤茶水提取物、70%乙醇提取物,采用K-B纸片法、微量稀释法测定不同藤茶提取物及其活性成分对创伤弧菌的抑菌环直径、最低抑菌浓度(MIC)、最低杀菌浓度(MBC)。利用酶标仪测定培养液在不同作用时间下的吸光度(A_(600))值,分析不同浓度藤茶醇提物对创伤弧菌生长特性的影响;利用酶标仪结合试剂盒法测定不同浓度藤茶醇提物作用创伤弧菌6 h后的细胞外碱性磷酸酶(AKP)、核酸及可溶性蛋白质的含量,分析其对菌体细胞壁及细胞膜的影响。结果 (1)藤茶水提物、藤茶70%醇提物、二氢杨梅素、没食子酸对创伤弧菌的抑菌环直径(mm)为18.68±0.71、21.45±0.63、20.16±0.49、19.59±0.67,MIC分别为5.00、0.63、1.25、2.50 mg/mL,MBC分别为5.00、0.63、2.50、5.00 mg/mL。(2)与空白组相比,藤茶70%醇提物MIC、2 MIC浓度组均能够显著抑制菌体生长和引起胞外AKP、核酸、蛋白质含量的显著提高(P<0.01)。结论 藤茶提取物均具有良好的抑杀创伤弧菌作用,以70%醇提物作用更强;其抗菌作用机制与破坏菌体细胞壁和细胞膜的完整性有关,可作为新型植物源抑菌剂进一步开发。Objective To study the antibacterial activity and mechanism of Ampelopsis grossedentata extract and its active components against Vibrio vulnificus in vitro, and provide experimental basis for its development and application. Methods Ampelopsis grossedentata water extract and 70% ethanol extract were prepared by heating reflux extraction. The inhibitory ring diameter, minimum inhibitory concentration(MIC) and minimum bactericidal concentration(MBC) of different Ampelopsis grossedentata extracts and their active components against Vibrio vulnificus were determined by K-B paper disk method and microdilution method. The A600value of the culture medium at different action time was measured by microplate reader to analyze the effect of different concentrations of alcoholic extract of Ampelopsis grossedentata on the growth characteristics of Vibrio vulnificus.The contents of extracellular alkaline phosphatase, nucleic acid and soluble protein of Vibrio vulnificus treated with different concentrations of Ampelopsis grossedentata ethanol extracts for 6 hours were determined by microplate reader and kit method, and its effects on cell wall and cell membrane were measured. Results(1) The inhibitory ring diameters of water extract of Ampelopsis grossedentata, 70% alcohol extract of Ampelopsis grossedentata, dihydromyricetin and gallic acid against Vibrio vulnificus were 18.68 ± 0.71 mm, 21.45 ± 0.63 mm,20.16±0.49 mm and 19.59±0.67 mm, MIC was 5.00, 0.63, 1.25 and 2.50 mg/m L, and MBC was 5.00, 0.63, 2.50 and5.00 mg/mL, respectively.(2) Compared with the blank group, the MIC and 2 MIC concentration groups of 70%ethanol extract of Ampelopsis grossedentata significantly inhibited the growth of bacteria and caused the significant increase of extracellular AKP, nucleic acid and protein content(P < 0.01). Conclusion The extracts of Ampelopsis grossedentata have good inhibitory effect on Vibrio vulnificus, and the 70% ethanol extract has a stronger effect. Its antibacterial mechanism is related to the destruction of the integr

关 键 词：藤茶 提取物 活性成分 创伤弧菌 抑菌活性 作用机制 

分 类 号：R285.5[医药卫生—中药学]