山大颜根、茎、叶提取物的体外抑菌活性评价及其作用机制研究
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篇名: 山大颜根、茎、叶提取物的体外抑菌活性评价及其作用机制研究
TITLE:
摘要: 目的:考察山大颜根、茎、叶3个部位提取物的体外抑菌活性,并探究其抑菌作用机制。方法:将山大颜根、茎、叶的95%醇提物均制备成质量浓度为100 mg/mL(以提取物计)的药液,并以青霉素钠(0.5 μg/mL)和硫酸链霉素(128 mg/mL)为阳性对照,采用平板打孔法测定不同药用部位提取物对金黄色葡萄球菌、大肠埃希菌、铜绿假单胞菌、枯草芽孢杆菌、藤黄微球菌、粪肠球菌等的抑菌作用。采用系统溶剂萃取法依次用石油醚、乙酸乙酯、正丁醇萃取山大颜抑菌活性部位,得到相应极性部位萃取物,制备成质量浓度均为100 mg/mL(以提取物计)药液后,考察其对上述6种菌的抑菌作用;并分别采用微量肉汤稀释法和琼脂培养基平板法测定其最低抑菌浓度(MIC)和最低杀菌浓度(MBC),筛选出具有抑菌作用的极性部位及对药物敏感的试验菌株。然后在0.5MIC和MIC药物浓度下绘制敏感菌的生长曲线(作用36 h,每4 h测定1次),检测菌悬液中碱性磷酸酶(AKP)和可溶性蛋白含量(作用10 h,每2 h测定1次),并以肉汤培养基代替提取物作为空白对照。结果:100 mg/mL山大颜根、茎和叶的醇提物对上述6种细菌均有较好的抑菌效果,其作用强弱为茎>叶>根;在山大颜茎的不同极性部位中,以乙酸乙酯部位的抑菌作用最强,尤其是对金黄色葡萄球菌,其抑菌圈直径为(38.93±0.12) mm,MIC、MBC均为0.39 mg/mL。山大颜茎乙酸乙酯部位可显著抑制金黄色葡萄球菌的增殖,菌悬液中碱性磷酸酶和蛋白质含量较空白对照显著升高(P<0.05)。结论:山大颜茎为抗菌活效部位,对常见的6种病原菌有不同程度的抑制效果;其中以山大颜茎的乙酸乙酯部位对金黄色葡萄球菌的抑制作用最为显著,能逐步破坏其细胞壁和细胞膜的完整性。
ABSTRACT: OBJECTIVE: To investigate in vitro antibacterial activity of the extracts from the root, stem and leaves of Psychotria rubra, and to investigate its mechanism of antibacterial effects. METHODS: 95% ethanol extracts from the root, stem and leaves of P. rubra were used to prepare solution with mass concentration of 100 mg/mL (calculated by extract). Using as penicillin sodium (0.5 mg/mL) and streptomycin sulfate (128 mg/mL) as positive control, plat stiletto method was used to determine antibacterial effects of the extracts from different parts of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Micrococcus luteus and Enterococcus faecalis. Using systematic solvent extraction method, the petroleum ether, ethyl acetate and n-butyl alcohol were used to extract antibacterial activity parts from P. rubra as ad to obtain the extracts of corresponding polar parts. After preparing 100 mg/mL drug solution (calculated by extract), antibacterial effects of above 6 kinds of bacteria were investigated. The micro-broth dilution method and agar culture medium plate method were used to determine minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), and screen polar parts with bacteriostasis and drug-sensitive strains. Under the concentration of 0.5MIC and MIC, the growth curves of sensitive bacteria were drawn (treated for 36 h, every 4 h); the contents of alkaline phosphatase (AKP) and soluble protein were detected in suspension (treated for 10 h, every 2 h); bouillon culture medium instead of the extract as blank control. RESULTS: 100 mg/mL ethanol extracts from the root, stem and leaves of P. rubra showed good antibacterial effect to above 6 kinds of bacteria, in descending order as stem>leaves>root. Among different polar parts from the stem of P. rubra, antibacterial effect of the ethyl acetate extract was best, especially for the S. aureus, the diameter of inhibition zone reached (38.93±0.12) mm, and both MIC and MBC were 0.39 mg/mL. The ethyl acetate extract could significantly inhibit the proliferation of S. aureus, and its alkaline phosphatase and protein content in suspension were increased significantly compared with blank control (P<0.05). CONCLUSIONS: The stem from P. rubra is effective antibacterial parts, and exhibit good antibacterial activity to 6 kinds of common pathogens. The ethyl acetate extract from the stem of P. rubra shows strongest antibacterial effect on S. aureus, and could gradually destroy the integrity of cell wall and cell membrane.
期刊: 2019年第30卷第1期
作者: 罗晓东,魏丽芳,钟昳,丘波,张超,邹浩元,张声源
AUTHORS: LUO Xiaodong,WEI Lifang,ZHONG Yi,QIU Bo,ZHANG Chao,ZOU Haoyuan,ZHANG Shengyuan
关键字: 山大颜;不同部位;体外抗菌活性;作用机制;最小抑菌浓度;最小杀菌浓度
KEYWORDS: Psychotria rubra; Different parts; Antibacterial activity in vitro; Mechanism of effect; Minimum inhibitory concentration; Minimum bactericidal concentration
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