摘要
目的探索传统润下药火麻仁Fructus Cannabis与人源肠道菌相互作用的特点。方法运用高效液相色谱(HPLC)法对火麻仁水煎液中主要的不饱和脂肪酸亚油酸和亚麻酸进行含量测定,采用固体与液体的体外厌氧培养方式,结合16Sr RNA技术分析火麻仁水煎液对人源肠道菌生长情况的影响,并采用HPLC法测定人源肠道菌对火麻仁水煎液中亚油酸与亚麻酸代谢的影响,同时对可能出现的新产物共轭亚麻酸及共轭亚油酸进行测定。结果火麻仁水煎液显著促进了变形菌门Proteobacteria的生长,表现为大肠志贺杆菌Escherichia-shigella显著增多(P<0.01)、脆弱拟杆菌Bacteroides显著减少(P<0.01),故拟杆菌门Bacteroidetes的生长受到了抑制,而火麻仁水煎液中含有的不饱和脂肪酸亚油酸和亚麻酸经人源肠道菌代谢后含量均降低,且有代谢产物共轭亚油酸和共轭亚麻酸的生成。结论火麻仁水煎液与人源肠道菌相互作用明显,药物能改变菌群结构,菌群能代谢药物成分,这种分析方法在一定程度上部分还原了药物口服途经肠道部位的物质代谢反应,为探明火麻仁在临床治疗中的物质基础提供了依据。
Objective To explore the characteristics of the interactions between traditional laxative medicine Cannabis Fructus and human gut microflora. Methods HPLC method was used to determine the content of the main unsaturated fatty acids linoleic acid and linolenic acid in Cannabis Fructus Decoction. At the same time, solid culture and liquid culture in vitro anaerobic culture method were combined with 16 S rRNA technology to analyze the interactions between Cannabis Fructus Decotion and human gut microflora.Moreover, the metabolits of linoleic acid and linolenic acid in Cannabis Fructus Decoction by human intestinal microflora were determined using HPLC method. At the same time, the possible conjugated linolenic acid and linoleic acid were determined. Results Cannabis Fructus Decoction promoted the growth of Proteobacteria significantly, which showed that Escherichia-shigella was significantly increased(P < 0.01), but the growth of Bacteroidetes was decreased(P < 0.01), and the content of unsaturated fatty acids linoleic acid and linolenic acid in Cannabis Fructus Decoction were reduced after being incubated with human intestinal bacteria, and the metabolites were conjugated linoleic acid and conjugated linolenic acid. Conclusion The interactions between Cannabis Fructus Decoction and human intestinal microflora are obvious. The Chinese medicine can change the structure of the gut microflora, and the gut microflora can metabolize the drug components. This analysis method partially restores the pharmacokinetics process of the oral administration drug in the human intestinal tract. It could provide a new insight of the mechanism research of Cannabis Fructus.
引文
[1]吴宿慧,郭媛媛,张神涛,等.火麻仁油在家蚕抗衰老实验中的性别差异的研究[J].中药药理与临床,2015,31(1):100-103.
[2]李寒冰,孙静雅,马永洁,等.火麻仁油及甾醇对家蚕寿命影响的观察[J].中医学报,2012,27(9):1145-1147.
[3]魏月媛,李理.火麻仁的功效及食用安全性研究进展[J].食品工业,2015,36(7):256-260.
[4]李根林,张颜语,李寒冰.复方地芬诺酯建立大鼠便秘模型及对肠道菌群的影响[J].中国组织工程研究,2016,20(49):7404-7410.
[5]李寒冰,吴宿慧,张颜语,等.基于肠道内环境调整的火麻仁润下作用的现代研究[J].中草药,2018,49(14):3334-3342.
[6]门薇,陈颖,杨庆,等.不同配伍戊己丸的黄连生物碱在人源肠道菌群中的代谢研究[J].中国中药杂志,2013,38(3):417-421.
[7]熊万娜,黄敏琪,梁健钦,等.离体人肠道菌群对柚皮苷的代谢动力学分析[J].中国实验方剂学杂志,2015,21(12):69-71.
[8]黄慧学,谭珍媛,邓家刚,等.人肠道菌群对芒果苷体外代谢转化的研究[J].中国中药杂志,2011,36(4):443-445.
[9]杨展.衰老肠道微生态的变化及干预措施研究[D].北京:中国人民解放军军事医学科学院,2017.
[10]孙翔宇,高贵田,段爱莉,等.多不饱和脂肪酸的研究进展[J].食品工业科技,2012,33(7):418-423.
[11]赵敏.亚油酸及亚油酸甲酯的抗炎作用研究[D].成都:西南交通大学,2012.
[12]李根林,曹亚蕊,吴宿慧,等.火麻仁油对衰老模型小鼠血脂水平及炎症、抗氧化相关指标的影响[J].中药药理与临床,2015,31(1):109-111.
[13]Kim K,Nam Y A,Kim H S,et al.α-Linolenic acid:Nutraceutical,pharmacological and toxicological evaluation[J].Food Chem Toxicol,2014,doi:10.1016/j.fct.2014.05.009.
[14]张金铭,张秀英,郝丽红,等.α-亚麻酸和花生四烯酸对Nrf 2和I相代谢酶CYP7A1表达的影响[J].黑龙江畜牧兽医,2016(21):1-4.
[15]Druart C,Neyrinck A M,Vlaeminck B,et al.Role of the lower and upper intestine in the production and absorption of gut microbiota-derived PUFA metabolites[J].PLoS One,2014,9(1):e87560.
[16]郜玉婷.利用紫苏油生物合成共轭亚麻酸的研究[D].大连:大连工业大学,2011.
[17]彭永健,许新德,张莉华,等.共轭亚麻酸的研究进展[J].中国食品添加剂,2015(1):165-170.
[18]Gorissen L,Raes K,Weckx S,et al.Production of conjugated linoleic acid and conjugated linolenic acid isomers by Bifidobacterium species[J].Appl Microbiol Biotechnol,2010,87(6):2257-2266.
[19]Kazunori K,Imamura G,Akashoshi A,et al.Genetically modified rapeseed oil containing cis-9,trans-11,cis-13-octadecatrienoic acid affects body fat mass and lipid metabolism in mice[J].J Agric Food Chem,2007,55(9):3741-3748.
[20]王丽敏,吕加平,王强.微生物生产共轭亚油酸的研究进展[J].中国乳品工业,2005,33(11):38-41.
[21]王中太,曹健,尹艳丽,等.共轭亚油酸的生物合成及相关酶基因的克隆与表达[J].化学与生物工程,2008,25(4):7-11.
