厚朴醇提物与远志醇提物配伍对大鼠粪便代谢物的影响
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摘要
目的:考察厚朴醇提物、远志醇提物及二者配伍对大鼠粪便代谢物的影响,分析其潜在的代谢通路,为探索厚朴缓解远志所致胃肠动力障碍的机制提供实验依据。方法:将40只SD雄性大鼠随机分为空白组,厚朴醇提物组(3. 50 g·kg~(-1)),远志醇提物组(1. 75 g·kg~(-1)),配伍组(3. 50 g·kg~(-1)+1. 75 g·kg~(-1)),连续灌胃给药3 d,收集大鼠末次给药24 h内的粪便,采用UPLC-Q-TOF-MS采集粪便代谢物数据,流动相乙腈(A)-0. 1%甲酸水溶液(B)梯度洗脱(0～0. 5 min,3%A;0. 5～12. 5 min,3%～70%A; 12. 5～13 min,70%～90%A; 13～16 min,90%～97%A; 16～17 min,97%～100%A; 17～22 min,100%A; 22～23 min,100%～3%A),电喷雾离子源,正、负离子模式数据采集范围m/z 50～1 200;借助Progenesis QI v2. 0,SIMCA-P 14. 0,SPSS 20. 0,MetaboAnalyst 4. 0等软件进行多元分析,筛选特征代谢标志物并分析其相关代谢通路。结果:共筛选出5-亚胺甲基四氢叶酸,L-3-羟基犬尿氨酸,7,8-二氢蝶酸等17个特征代谢标志物,其相关代谢通路为不饱和脂肪酸的生物合成、亚油酸代谢、维生素B_6代谢、叶酸生物合成等。结论:厚朴缓解远志所致胃肠动力障碍的作用机制可能与嘌呤代谢、叶酸生物合成、色氨酸代谢、初级胆汁酸生物合成有关。
Objective: To investigate the effect of alcohol extract of Magnoliae Officinalis Cortex,alcohol extract of Polygalae Radix and their compatibility on fecal metabolites of rats, analyze its potential metabolic pathways,and provide experimental basis for exploring the possible mechanism of Magnoliae Officinalis Cortex relieving gastrointestinal motility disorders induced by Polygalae Radix. Method: Forty male SD rats were randomly divided into the normal group,alcohol extract of Magnoliae Officinalis Cortex group( 3. 50 g·kg~(-1)),alcohol extract of Polygalae Radix group( 1. 75 g·kg~(-1)) and compatibility group( 3. 5 g·kg~(-1) of alcohol extract of Magnoliae Officinalis Cortex + 1. 75 g·kg~(-1) of alcohol extract of Polygalae Radix). Fecal samples were collected within 24 h after continuous gavage for 3 days. The fecal metabolites in each group was detected by ultra-high performance liquid chromatography-quadrupole-time of flight-mass spectrometry( UPLC-Q-TOF-MS), mobile phase was acetonitrile-0. 1% formic acid solution for gradient elution,data collection range was m/z 50~(-1) 200 under positive and negative ion mode of electrospray ionization. The characteristic biomarkers and corresponding metabolic pathways were analyzed or screened by Progenesis QI v2. 0, SIMCA-P 14. 0, SPSS 20. 0,MetaboAnalyst 4. 0 and other softwares. Result: A total of 17 characteristic metabolic markers were screened out,including 5-formiminotetrahydrofolic acid, L-3-hydroxykynurenine, 7, 8-dihydropteroic acid, etc. The main related pathways included biosynthesis of unsaturated fatty acids, linoleic acid metabolism, vitamin B_6 metabolism,etc. Conclusion: The mechanism of Magnoliae Officinalis Cortex relieving gastrointestinal motility disorders induced by Polygalae Radix may be related to purine metabolism, folate biosynthesis, tryptophan metabolism and primary bile acid biosynthesis.
Objective: To investigate the effect of alcohol extract of Magnoliae Officinalis Cortex,alcohol extract of Polygalae Radix and their compatibility on fecal metabolites of rats, analyze its potential metabolic pathways,and provide experimental basis for exploring the possible mechanism of Magnoliae Officinalis Cortex relieving gastrointestinal motility disorders induced by Polygalae Radix. Method: Forty male SD rats were randomly divided into the normal group,alcohol extract of Magnoliae Officinalis Cortex group( 3. 50 g·kg~(-1)),alcohol extract of Polygalae Radix group( 1. 75 g·kg~(-1)) and compatibility group( 3. 5 g·kg~(-1) of alcohol extract of Magnoliae Officinalis Cortex + 1. 75 g·kg~(-1) of alcohol extract of Polygalae Radix). Fecal samples were collected within 24 h after continuous gavage for 3 days. The fecal metabolites in each group was detected by ultra-high performance liquid chromatography-quadrupole-time of flight-mass spectrometry( UPLC-Q-TOF-MS), mobile phase was acetonitrile-0. 1% formic acid solution for gradient elution,data collection range was m/z 50~(-1) 200 under positive and negative ion mode of electrospray ionization. The characteristic biomarkers and corresponding metabolic pathways were analyzed or screened by Progenesis QI v2. 0, SIMCA-P 14. 0, SPSS 20. 0,MetaboAnalyst 4. 0 and other softwares. Result: A total of 17 characteristic metabolic markers were screened out,including 5-formiminotetrahydrofolic acid, L-3-hydroxykynurenine, 7, 8-dihydropteroic acid, etc. The main related pathways included biosynthesis of unsaturated fatty acids, linoleic acid metabolism, vitamin B_6 metabolism,etc. Conclusion: The mechanism of Magnoliae Officinalis Cortex relieving gastrointestinal motility disorders induced by Polygalae Radix may be related to purine metabolism, folate biosynthesis, tryptophan metabolism and primary bile acid biosynthesis.
引文
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[11]唐冰雯,李俊健,毋福海,等.甘遂-甘草配伍对大鼠肾脏毒性代谢组学的影响[J].中国实验方剂学杂志,2015,21(9):88-92.
[12]高鹏飞,刘卫红,吴俊珠,等.代谢组学在中医药研究中的应用进展[J].中国实验方剂学杂志,2011,17(21):284-288.
[13]SUN B,WANG X B,CAO R L,et al. NMR-based metabonomics study on the effect of Gancao in the attenuation of toxicity in rats induced by Fuzi[J]. J Ethnopharmacol,2016,193:617-626.
[14]章斌,娄子洋.液质联用技术在中药代谢物及代谢组学研究中的应用分析[J].第二军医大学学报,2011,32(6):658-662.
[15]Williams M D,XIAN L,Huso T,et al. The fecal metabolome in hmga1 transgenic mice with polyposis:evidence for potential screen for early detection of precursor lesions in colorectal cancer[J]. J Proteome Res,2016,15(12):4176-4187.
[16]YU M,JIA H M,ZHOU C,et al. Urinary and fecal metabonomics study of the protective effect of ChaihuShu-Gan-San on antibiotic-induced gut microbiota dysbiosis in rats[J]. Sci Rep,2017,doi:10. 1038/srep46551.
[17]杨瑞,唐思,董晓茜,等.基于UPLC-Q-TOF-MS分析雷公藤配伍甘草治疗肾病综合征大鼠的尿液代谢组学[J].中国实验方剂学杂志,2018,24(12):150-156.
[18]郭慧,崔扬,王秋红,等.基于代谢组学技术的中药复方研究近况[J].中国实验方剂学杂志,2017,23(1):213-219.
[19]黄立华,王建,吴明权,等.远志、厚朴配伍对其活性成分在大鼠肠段吸收的影响[J].中成药,2015,37(4):739-745.
[20]罗凤娟,王建,马骁,等.远志-厚朴配伍对厚朴酚、和厚朴酚胃肠代谢的影响[J].中国实验方剂学杂志,2013,19(7):154-158.
[21]Ho C Y,Miller K V,Savaiano D A,et al. Absorption and metabolism of orally administered purines in fed and fasted rats[J]. J Nutr,1979,109(8):1377-1382.
[22]Witte D P,Wiginton D A,Hutton J J,et al. Coordinate developmental regulation of purine catabolic enzyme expression in gastrointestinal and postimplantation reproductive tracts[J]. J Cell Biol,1991,115(1):179-190.
[23]朱舜时,夏书军,陈晓宇,等.叶酸治疗萎缩性胃炎并干预胃癌生成的分子生物学机制[J].胃肠病学,2002,7(2):86-89.
[24]李小兰,江敏,阮征,等.色氨酸及其代谢产物5-HT对肠道功能的作用综述[J].食品安全质量检测学报,2014,5(7):1997-2002.
[25]丁健华,傅传刚,赵荣华.五羟色胺在胃肠道功能性疾病中的研究现状[J].世界华人消化杂志,2005,13(20):2405-2408.
[26]张卫卫.半夏厚朴汤及厚朴主要成分促胃肠运动、抗焦虑和抗抑郁作用的研究[D].沈阳:中国医科大学,2004.
[27]AlméB,Bremmelgaard A,Sj9vall J,et al. Analysis of metabolic profiles of bile acids in urine using a lipophilic anion exchanger and computerized gas-liquid chromatorgaphy-mass spectrometry[J]. J Lipid Res,1977,18(3):339-362.
[28]徐运杰,汪前红.胆汁酸的作用与分子调控机制[J].饲料博览,2015,36(2):21-25.
[29]Armstrong D N,Krenz H K,Modlin I M,et al. Bile salt inhibition of motility in the isolated perfused rabbit terminal ileum[J]. Gut,1993,34(4):483-488.
[30]Penagini R,Misiewicz J J,Frost P G. Effect of jejunal infusion of bile acids on small bowel transit and fasting jejunal motility in man[J]. Gut,1988,29:789-794.
[31]Jr S W,Shiff S,Cohen S. Effect of deoxycholic acid on colonic motility in the rabbit[J]. Am J Physiol,1980,238(4):G321-G325.
[32]周军.基于肝肠循环的“王氏益灵丸”神阙穴外治肝硬化肝源性腹泻的临床研究[D].上海:上海中医药大学,2016.
[33]王会敏,王正平,董旻岳.胆汁酸代谢与调控研究进展[J].国际消化病杂志,2010,30(2):79-82.
[34]张久聪,聂青和.胆汁酸代谢及相关进展[J].胃肠病学和肝病学杂志,2008,17(11):953-956.
[35]孟宪元,尹珉,王学江.厚朴对低温条件下大鼠胆汁分泌影响的初步观察[J].北京中医,2005,24(3):180-181.
[36]陆茵,张大方.中药药理学[M].北京:人民卫生出版社,2015:126.
[2]马洪伟,付文亮,钟美蓉,等.远志药理活性研究进展[J].承德医学院学报,2010,27(2):196-198.
[3]田徽.生远志的胃肠毒性物质基础及蜜炙减毒部分机理研究[D].成都:成都中医药大学,2006.
[4]武云.生远志与蜜远志的毒理及其对胃肠运动影响的实验研究[D].成都:成都中医药大学,2005.
[5]江娟,王建,李达,等.远志与厚朴不同煎煮时间及不同配比对小鼠胃肠运动的影响[J].西安交通大学学报:医学版,2009,30(3):380-382.
[6]马骁.厚朴汁炙远志减副存效及化学成分研究[D].成都:成都中医药大学,2013.
[7]唐丹霞,王建,袁奕,等.厚朴汁炙远志含药血清对胃Cajal间质细胞活力及n NOS表达的影响[J].中药药理与临床,2013,29(6):100-103.
[8]唐丹霞,王建,郑新光,等.厚朴汁炙远志炮制品对胃间质细胞活力影响的探讨[J].中药与临床,2014,5(2):63-65,68.
[9]DONG H,YAN G L,HAN Y,et al. UPLC-Q-TOF/MSbased metabolomic studies on the toxicity mechanisms of traditional Chinese medicine Chuanwu and the detoxification mechanisms of Gancao,Baishao,and Ganjiang[J]. Chin J Nat Med,2015,13(9):687-698.
[10]马致洁,章从恩,唐进法,等.雷公藤配伍甘草降低肝毒性的代谢通路探讨[J].药学学报,2017,52(7):1077-1084.
[11]唐冰雯,李俊健,毋福海,等.甘遂-甘草配伍对大鼠肾脏毒性代谢组学的影响[J].中国实验方剂学杂志,2015,21(9):88-92.
[12]高鹏飞,刘卫红,吴俊珠,等.代谢组学在中医药研究中的应用进展[J].中国实验方剂学杂志,2011,17(21):284-288.
[13]SUN B,WANG X B,CAO R L,et al. NMR-based metabonomics study on the effect of Gancao in the attenuation of toxicity in rats induced by Fuzi[J]. J Ethnopharmacol,2016,193:617-626.
[14]章斌,娄子洋.液质联用技术在中药代谢物及代谢组学研究中的应用分析[J].第二军医大学学报,2011,32(6):658-662.
[15]Williams M D,XIAN L,Huso T,et al. The fecal metabolome in hmga1 transgenic mice with polyposis:evidence for potential screen for early detection of precursor lesions in colorectal cancer[J]. J Proteome Res,2016,15(12):4176-4187.
[16]YU M,JIA H M,ZHOU C,et al. Urinary and fecal metabonomics study of the protective effect of ChaihuShu-Gan-San on antibiotic-induced gut microbiota dysbiosis in rats[J]. Sci Rep,2017,doi:10. 1038/srep46551.
[17]杨瑞,唐思,董晓茜,等.基于UPLC-Q-TOF-MS分析雷公藤配伍甘草治疗肾病综合征大鼠的尿液代谢组学[J].中国实验方剂学杂志,2018,24(12):150-156.
[18]郭慧,崔扬,王秋红,等.基于代谢组学技术的中药复方研究近况[J].中国实验方剂学杂志,2017,23(1):213-219.
[19]黄立华,王建,吴明权,等.远志、厚朴配伍对其活性成分在大鼠肠段吸收的影响[J].中成药,2015,37(4):739-745.
[20]罗凤娟,王建,马骁,等.远志-厚朴配伍对厚朴酚、和厚朴酚胃肠代谢的影响[J].中国实验方剂学杂志,2013,19(7):154-158.
[21]Ho C Y,Miller K V,Savaiano D A,et al. Absorption and metabolism of orally administered purines in fed and fasted rats[J]. J Nutr,1979,109(8):1377-1382.
[22]Witte D P,Wiginton D A,Hutton J J,et al. Coordinate developmental regulation of purine catabolic enzyme expression in gastrointestinal and postimplantation reproductive tracts[J]. J Cell Biol,1991,115(1):179-190.
[23]朱舜时,夏书军,陈晓宇,等.叶酸治疗萎缩性胃炎并干预胃癌生成的分子生物学机制[J].胃肠病学,2002,7(2):86-89.
[24]李小兰,江敏,阮征,等.色氨酸及其代谢产物5-HT对肠道功能的作用综述[J].食品安全质量检测学报,2014,5(7):1997-2002.
[25]丁健华,傅传刚,赵荣华.五羟色胺在胃肠道功能性疾病中的研究现状[J].世界华人消化杂志,2005,13(20):2405-2408.
[26]张卫卫.半夏厚朴汤及厚朴主要成分促胃肠运动、抗焦虑和抗抑郁作用的研究[D].沈阳:中国医科大学,2004.
[27]AlméB,Bremmelgaard A,Sj9vall J,et al. Analysis of metabolic profiles of bile acids in urine using a lipophilic anion exchanger and computerized gas-liquid chromatorgaphy-mass spectrometry[J]. J Lipid Res,1977,18(3):339-362.
[28]徐运杰,汪前红.胆汁酸的作用与分子调控机制[J].饲料博览,2015,36(2):21-25.
[29]Armstrong D N,Krenz H K,Modlin I M,et al. Bile salt inhibition of motility in the isolated perfused rabbit terminal ileum[J]. Gut,1993,34(4):483-488.
[30]Penagini R,Misiewicz J J,Frost P G. Effect of jejunal infusion of bile acids on small bowel transit and fasting jejunal motility in man[J]. Gut,1988,29:789-794.
[31]Jr S W,Shiff S,Cohen S. Effect of deoxycholic acid on colonic motility in the rabbit[J]. Am J Physiol,1980,238(4):G321-G325.
[32]周军.基于肝肠循环的“王氏益灵丸”神阙穴外治肝硬化肝源性腹泻的临床研究[D].上海:上海中医药大学,2016.
[33]王会敏,王正平,董旻岳.胆汁酸代谢与调控研究进展[J].国际消化病杂志,2010,30(2):79-82.
[34]张久聪,聂青和.胆汁酸代谢及相关进展[J].胃肠病学和肝病学杂志,2008,17(11):953-956.
[35]孟宪元,尹珉,王学江.厚朴对低温条件下大鼠胆汁分泌影响的初步观察[J].北京中医,2005,24(3):180-181.
[36]陆茵,张大方.中药药理学[M].北京:人民卫生出版社,2015:126.
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