Bioactivities of Alchemilla mollis, Alchemilla persica and Their Active Constituents

Abstract The aim of this study was to determine antimicrobial activities of Alchemilla mollis, Alchemilla persica as well as ellagic acid and miquelianin against Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, Candida albicans by using microbroth dilution method and anti-inflammatory activity by using human red blood cell (HRBC) membrane stabilization method. Microbroth dilution method was used to determine the antimicrobial activities. Extracts possessed activity having MIC values of 2.5-5-10mg/ mL, compounds possessed activity having MIC values of 1.25-2.5-4-5mg/mL. A.mollis aerial parts displayed the highest anti-inflammatory activity (IC50=1.22±0.07mg/mL). Ellagic acid and miquelianin were also determined as anti-inflammatory agents with 0.57±0.01mg/mL and 1.23±0.02mg/mL IC50 values, respectively. Total phenolic content and tannin content of the A.mollis and A.persica were determined as 357.00±75.80mg, 282.50±28.70mg PGE/g plant material and 18.02%, 18.63% respectively according to the method described by European Pharmacopoeia. Ellagic acid, miquelianin and catechin were analyzed by HPLC. The highest catechin content was detected in A.persica roots (6.69±0.05g/100g plant material). A.mollis aerial parts contain higher miquelianin (0.39±0.02g/plant material) and ellagic acid (1.56±0.01g/ plant material) than A.persica.

摘要 本研究旨在通过微量肉汤稀释法，检测斗篷草（Alchemilla mollis）、波斯斗篷草（Alchemilla persica）、鞣花酸（ellagic acid）及miquelianin（miquelianin）对金黄色葡萄球菌（Staphylococcus aureus）、粪肠球菌（Enterococcus faecalis）、枯草芽孢杆菌（Bacillus subtilis）、大肠埃希菌（Escherichia coli）、铜绿假单胞菌（Pseudomonas aeruginosa）及白色念珠菌（Candida albicans）的抗菌活性，并采用人红细胞（human red blood cell, HRBC）膜稳定法测定其抗炎活性。
本研究采用微量肉汤稀释法开展抗菌活性检测。结果显示，供试提取物的最低抑菌浓度（minimum inhibitory concentration, MIC）为2.5 mg/mL、5 mg/mL及10 mg/mL，单体化合物的MIC为1.25 mg/mL、2.5 mg/mL、4 mg/mL及5 mg/mL。斗篷草地上部位的抗炎活性最优，其半数抑制浓度（half maximal inhibitory concentration, IC50）为1.22±0.07mg/mL。鞣花酸与miquelianin同样具备抗炎活性，对应的IC50值分别为0.57±0.01mg/mL及1.23±0.02mg/mL。
按照欧洲药典（European Pharmacopoeia）记载的分析方法，本研究测定了斗篷草与波斯斗篷草的总酚含量及鞣质含量：斗篷草的总酚含量为357.00±75.80mg PGE/g植物材料，鞣质含量为18.02%；波斯斗篷草的总酚含量为282.50±28.70mg PGE/g植物材料，鞣质含量为18.63%。
采用高效液相色谱法（High Performance Liquid Chromatography, HPLC）对鞣花酸、miquelianin及儿茶素（catechin）进行了含量分析。结果检测到波斯斗篷草根中的儿茶素含量最高，为6.69±0.05g/100g植物材料。斗篷草地上部位的miquelianin及鞣花酸含量均高于波斯斗篷草，其中miquelianin含量为0.39±0.02g/植物材料，鞣花酸含量为1.56±0.01g/植物材料。