多叶棘豆体外抗菌作用部位的筛选研究

孟根小++奥?乌力吉++霍万学++王秀兰++王欢

摘要：目的 对比研究多叶棘豆不同提取物对临床常见6种致病菌体外抗菌作用，筛选抗菌活性部位，初步判断药效物质种类。方法 以多叶棘豆不同溶剂（石油醚、二氯甲烷、乙酸乙酯、正丁醇）提取物为试验药物，采用微孔板二倍稀释TTC法和琼脂平板稀释法测定最低抑菌浓度（MIC），液体转染法测定最小杀菌浓度（MBC）。采用分光光度法测定总黄酮含量。结果 多叶棘豆5种提取物均有体外抗菌作用，其中乙酸乙酯提取物抗菌作用最强，对最敏感菌（金黄色葡萄球菌）的MIC和MBC分别为0.02 mg/mL和1.25 mg/mL；其次为正丁醇提取物和石油醚提取物；剩余物的作用最弱。各提取物的总黄酮含量大小与抗菌活性强弱顺序一致，即乙酸乙酯提取物>正丁醇提取物>石油醚提取物>二氯甲烷提取物>剩余物。结论 多叶棘豆体外抗菌作用与黄酮类成分密切相关。

关键词：多叶棘豆；提取物；体外抗菌；作用部位

DOI：10.3969/j.issn.1005-5304.2016.12.013

中图分类号：R284.1 文献标识码：A 文章编号：1005-5304（2016）12-0051-04

Study on Screening for Antibacterial Active Site of Oxytropis Myriophylla in Vitro MENG Gen-xiao1，2， AO Wuliji2， HUO Wan-xue2， WANG Xiu-lan2， WANG Huan2 （1. College of Agronomy and Biotechnology， China Agricultural University， Beijing 100193， China； 2. College of Traditional Mongolian Medicine， Inner Mongolia University for the Nationalities， Tongliao 028300， China）

Abstract： Objective To compare the antibacterial activity of the different extracts from Oxytropis Myriophylla against 6 common pathogens in vitro； To select the best active site of Oxytropis Myriophylla with antibacterial activity； To preliminarily determine the material basis for efficacy category. Methods 4 different solvent （petroleum ether， dichloromethane， ethyl acetate and normal butanol） extracts of Oxytropis Myriophylla were selected as experimental medicine. MIC was determined by microtiter plate double dilution TTC method and agar disk dilution method. MBC were tested by liquid infection method. Spectrophotometric method was adopted to determine the contents of total flavonoid in the samples. Results All extracts had the antibacterial activity in vitro. But the ethyl acetate extract was the best one that MIC and MBC against one the most sensitive pathogen （Staph. aureus） were 0.02 mg/mL and 1.25mg/mL. Followed by normal butanol extract and petroleum ether extract， the remainder had the lowest activity. Total flavonoid contents of the extracts were exactly the same order with antibacterial effects of them：that ethyl acetate extract > normal butanol extract > petroleum ether extract > dichloromethane extract > remainder. Conclusion The antibacterial activity of Oxytropis Myriophylla is closely related to flavonoid.

Key words： Oxytropis Myriophylla； solvent extracts； antibacterial activity in vitro； active site

多叶棘豆为豆科植物狐尾藻棘豆Oxytropis myriophylla（Pall.）DC.的干燥地上部分，蒙药名为“那布其热哈格·乌如吐扎”，是蒙医常用杀“粘”药物之一，具有杀粘、清热、燥黄水、愈伤、生肌、合脉止

基金项目：国家科技支撑计划（2012BA128B00）

血、消肿、软便的功效[1-2]。多叶棘豆含有黄酮类[3-4]、木酚素类[5]、三萜皂苷[6]和生物碱类[7]等多种成分。因此，化学成分的探讨成为该药材的研究热点。但关于杀“粘”作用，只有对白假丝酵母和大肠杆菌的抗菌研究[8-9]。为了进一步评价多叶棘豆的杀“粘”作用、筛选最佳活性部位，本实验研究该药不同提取物对临床常见6种致病菌的体外抗菌作用，并基于笔者从该药提取的化合物多为黄酮类[3]，测定各部位的总黄酮含量，初步判断抑菌活性物质。 ……