基于代谢组学研究厚朴“发汗”前后对胃肠动力障碍大鼠的影响
|
摘要
基于代谢组学方法,对厚朴"发汗"前后治疗胃肠动力障碍大鼠的血清代谢物进行对比,研究厚朴"发汗"增效减毒的作用机制。采用L-精氨酸腹腔注射建立胃肠动力障碍大鼠模型,以大鼠胃内色素相对残留率和小肠推进比作为药效学指标,采用LC-MS的代谢组学技术,并结合主成分分析(PCA)、偏最小二乘判别分析(PLS-DA)和正交校正的偏最小二乘判别分析(OPLS-DA)等方法筛选并鉴定与胃肠动力障碍相关的生物标志物,利用MetPA数据库分析相关代谢通路。结果表明厚朴不管发汗与否,均能改善胃肠动力障碍,且"发汗"厚朴作用强于"未发汗"厚朴组。实验各组的代谢物能够显著区分,发现并初步鉴定了15个差异代谢物及17条相关通路。厚朴起效的机制可能是通过提高丙氨酸、天门冬氨酸和谷氨酸代谢通路中L-谷氨酸胺的含量,保护胃肠道屏障而改善胃肠动力障碍;厚朴"发汗"前通过牛磺酸和亚牛磺酸代谢,初级胆汁酸生物合成通路而降低牛磺酸,提高甘氨鹅脱氧胆酸含量,可能会加大肝肾损伤几率;而厚朴"发汗"后,通过提高色氨酸通路中L-色氨酸和5-羟色胺含量来进一步增强胃肠动力,并避免了有害代谢物产生从而达到增效减毒的作用。
Based on metabolomics,the effect of Magnolia officinalis before and after " sweating" on gastrointestinal motility disorder( rat) was compared. To study the mechanism of M. officinalis " sweating" increased the efficacy and reduced the toxicity. The rat model of gastrointestinal motility disorder was established by intraperitoneal injection of L-arginine. Pharmacodynamic indexes were relative residual rate of gastric pigment and intestinal propulsion ratio in rats. LC-MS metabolomics and multivariate statistical analysis were used to screen and identify biomarkers associated with gastrointestinal motility disorders,and MetPA database was used to analyze related metabolic pathways. The results showed that M. officinalis could improve gastrointestinal motility disorder whether it " sweating" or not,and the effect of " sweating" M. officinalis was stronger than that of " no sweating" M. officinalis. The metabolites of the experimental groups could be distinguished distinctly,and 15 different compounds and 17 related pathways were identified preliminarily. The mechanism of M. officinalis might be to improve gastrointestinal motility disorder by increasing the content of L-glutamate in the metabolic pathway of alanine,aspartate and glutamate and protecting gastrointestinal barrier. Before " sweating",M. officinalis could reduce taurine through metabolism of taurine and taurine and biosynthetic pathway of primary bile acid,increase the content of deoxycholic acid in glycine goose,and increase the risk of liver and kidney injury. After " sweating",M. officinalis could enhance gastrointestinal motility by increasing the contents of L-tryptophan and serotonin in the tryptophan pathway,and avoid the production of harmful metabolites to achieve synergistic and detoxifying effect.
Based on metabolomics,the effect of Magnolia officinalis before and after " sweating" on gastrointestinal motility disorder( rat) was compared. To study the mechanism of M. officinalis " sweating" increased the efficacy and reduced the toxicity. The rat model of gastrointestinal motility disorder was established by intraperitoneal injection of L-arginine. Pharmacodynamic indexes were relative residual rate of gastric pigment and intestinal propulsion ratio in rats. LC-MS metabolomics and multivariate statistical analysis were used to screen and identify biomarkers associated with gastrointestinal motility disorders,and MetPA database was used to analyze related metabolic pathways. The results showed that M. officinalis could improve gastrointestinal motility disorder whether it " sweating" or not,and the effect of " sweating" M. officinalis was stronger than that of " no sweating" M. officinalis. The metabolites of the experimental groups could be distinguished distinctly,and 15 different compounds and 17 related pathways were identified preliminarily. The mechanism of M. officinalis might be to improve gastrointestinal motility disorder by increasing the content of L-glutamate in the metabolic pathway of alanine,aspartate and glutamate and protecting gastrointestinal barrier. Before " sweating",M. officinalis could reduce taurine through metabolism of taurine and taurine and biosynthetic pathway of primary bile acid,increase the content of deoxycholic acid in glycine goose,and increase the risk of liver and kidney injury. After " sweating",M. officinalis could enhance gastrointestinal motility by increasing the contents of L-tryptophan and serotonin in the tryptophan pathway,and avoid the production of harmful metabolites to achieve synergistic and detoxifying effect.
引文
[1]陈茹,陈成,杨兴鑫,等.中药“发汗”炮制法的现代研究进展[J].中草药,2018,49(2):489.
[2]刘红亮,晏仁义,郭健,等.厚朴“发汗”前后药材颜色及气味差异的数值化研究[J].中国中药杂志,2013,38(1):45.
[3]余盛贤,张春霞,陈承瑜,等.“发汗”对厚朴质量的影响[J].中国中药杂志,2010,35(14):1831.
[4]尹爱武,田润,黄国文,等.厚朴“发汗”对大鼠胃动力和抗溃疡作用研究[J].中国实验方剂学杂志,2012,18(16):212.
[5]郭彬彬,于庆生,沈毅,等.中医药对应激性胃肠动力障碍的治疗研究进展[J].中医药临床杂志,2018,30(3):578.
[6]巢蕾,曹雨诞,陈佩东,等.厚朴对胃肠动力障碍作用的研究[J].中国医药导报,2018,15(13):31.
[7]Mélanie P,Pierre D.Biological activity and toxicity of the Chinese herb Magnolia officinalis Rehder&E.Wilson(Houpo)and its constituents[J].J Zhejiang Univ Sci B,2017,18(3):194.
[8]Sangster T,Major H,Plumb R,et al.A pragmatic and readily implemented quality control strategy for HPLC-MS and GC-MS-based metabonomic analysis[J].Analyst,2006,131(10):1075.
[9]Want E J,Masson P,Michopoulos F,et al.Global metabolic profiling of animal and human tissues via UPLC-MS[J].Nat Protoc,2013,8:17.
[10]Smith C A,Want E J,O'Maille G,et al.XCMS:processing mass spectrometry data for metabolite profiling using nonlinear peak alignment,matching,and identification[J].Anal Chem,2006,78:779.
[11]Trygg J,Wold S.Orthogonal projections to latent structures(O-PLS)[J].J Chemometr,2002,16:119.
[12]王满才.基于RAS系统及NO/c GMP通路研究L-Arg盐溶液制备胃肠动力障碍大鼠模型的可行性[D].兰州:兰州大学,2016.
[13]Zhang Y,Li H,Hu T,et al.Metabonomic profiling in study hepatoprotective effect of polysaccharides from Flammulina velutipes on carbon tetrachloride-induced acute liver injury rats using GC-MS[J].Int J Biol Macromol,2018,110:285.
[14]沈丽娟,王倩,吴锡平,等.谷氨酰胺对脓毒症大鼠急性肠黏膜损伤的保护作用[J].内科急危重症杂志,2018,24(3):253.
[15]张晓松,马琪,文艳巧,等.苦豆草治疗大肠湿热证大鼠血清代谢组学研究[J].药学学报,2018,53(1):111.
[16]刘彦玲,张丛,王强,等.糖尿病肾病代谢组学差异代谢物的研究进展[J].药物评价研究,2018,10(21):1.
[17]张燕军,罗庆锋,张静,等.甘氨鹅去氧胆酸对大鼠肝窦内皮细胞凋亡的影响[J].胃肠病学,2006(5):281.
[18]Messineo A M,Gineste C,Sztal T E,et al.L-tyrosine supplementation does not ameliorate skeletal muscle dysfunction in zebrafish and mouse models of dominant skeletal muscleα-actin nemaline myopathy[J].Sci Rep,2018,8(1):11490.
[19]Layunta E,Latorre E,Forcen R,et al.NOD1 downregulates intestinal serotonin transporter and interacts with other pattern recognition receptors[J].J Cell Physiol,2018,233:4183.
[20]Swami T,Weber H C.Updates on the biology of serotonin and tryptophan hydroxylase[J].Curr Opin Endocrinol Diabetes Obes,2018,25:12.
[21]耿建军.厚朴排气汤对腹外科患者术后胃肠道功能障碍的影响[J].光明中医,2018,33(8):1120.
[22]陈建飞,曹羽.厚朴三物汤加半夏高位灌肠治疗胃肠道术后肠麻痹的临床体会[J].中国临床医生,2004(9):46.
[23]张广安.桂枝厚朴杏子汤加味治疗胃肠道功能紊乱60例[J].现代中医药,2013,33(2):30.
[24]唐荣伟,李德科,唐玲,等.厚朴温中汤加味配伍吗丁啉对功能性消化不良血浆胃肠激素的影响[J].中国实验方剂学杂志,2015,21(13):174.
[2]刘红亮,晏仁义,郭健,等.厚朴“发汗”前后药材颜色及气味差异的数值化研究[J].中国中药杂志,2013,38(1):45.
[3]余盛贤,张春霞,陈承瑜,等.“发汗”对厚朴质量的影响[J].中国中药杂志,2010,35(14):1831.
[4]尹爱武,田润,黄国文,等.厚朴“发汗”对大鼠胃动力和抗溃疡作用研究[J].中国实验方剂学杂志,2012,18(16):212.
[5]郭彬彬,于庆生,沈毅,等.中医药对应激性胃肠动力障碍的治疗研究进展[J].中医药临床杂志,2018,30(3):578.
[6]巢蕾,曹雨诞,陈佩东,等.厚朴对胃肠动力障碍作用的研究[J].中国医药导报,2018,15(13):31.
[7]Mélanie P,Pierre D.Biological activity and toxicity of the Chinese herb Magnolia officinalis Rehder&E.Wilson(Houpo)and its constituents[J].J Zhejiang Univ Sci B,2017,18(3):194.
[8]Sangster T,Major H,Plumb R,et al.A pragmatic and readily implemented quality control strategy for HPLC-MS and GC-MS-based metabonomic analysis[J].Analyst,2006,131(10):1075.
[9]Want E J,Masson P,Michopoulos F,et al.Global metabolic profiling of animal and human tissues via UPLC-MS[J].Nat Protoc,2013,8:17.
[10]Smith C A,Want E J,O'Maille G,et al.XCMS:processing mass spectrometry data for metabolite profiling using nonlinear peak alignment,matching,and identification[J].Anal Chem,2006,78:779.
[11]Trygg J,Wold S.Orthogonal projections to latent structures(O-PLS)[J].J Chemometr,2002,16:119.
[12]王满才.基于RAS系统及NO/c GMP通路研究L-Arg盐溶液制备胃肠动力障碍大鼠模型的可行性[D].兰州:兰州大学,2016.
[13]Zhang Y,Li H,Hu T,et al.Metabonomic profiling in study hepatoprotective effect of polysaccharides from Flammulina velutipes on carbon tetrachloride-induced acute liver injury rats using GC-MS[J].Int J Biol Macromol,2018,110:285.
[14]沈丽娟,王倩,吴锡平,等.谷氨酰胺对脓毒症大鼠急性肠黏膜损伤的保护作用[J].内科急危重症杂志,2018,24(3):253.
[15]张晓松,马琪,文艳巧,等.苦豆草治疗大肠湿热证大鼠血清代谢组学研究[J].药学学报,2018,53(1):111.
[16]刘彦玲,张丛,王强,等.糖尿病肾病代谢组学差异代谢物的研究进展[J].药物评价研究,2018,10(21):1.
[17]张燕军,罗庆锋,张静,等.甘氨鹅去氧胆酸对大鼠肝窦内皮细胞凋亡的影响[J].胃肠病学,2006(5):281.
[18]Messineo A M,Gineste C,Sztal T E,et al.L-tyrosine supplementation does not ameliorate skeletal muscle dysfunction in zebrafish and mouse models of dominant skeletal muscleα-actin nemaline myopathy[J].Sci Rep,2018,8(1):11490.
[19]Layunta E,Latorre E,Forcen R,et al.NOD1 downregulates intestinal serotonin transporter and interacts with other pattern recognition receptors[J].J Cell Physiol,2018,233:4183.
[20]Swami T,Weber H C.Updates on the biology of serotonin and tryptophan hydroxylase[J].Curr Opin Endocrinol Diabetes Obes,2018,25:12.
[21]耿建军.厚朴排气汤对腹外科患者术后胃肠道功能障碍的影响[J].光明中医,2018,33(8):1120.
[22]陈建飞,曹羽.厚朴三物汤加半夏高位灌肠治疗胃肠道术后肠麻痹的临床体会[J].中国临床医生,2004(9):46.
[23]张广安.桂枝厚朴杏子汤加味治疗胃肠道功能紊乱60例[J].现代中医药,2013,33(2):30.
[24]唐荣伟,李德科,唐玲,等.厚朴温中汤加味配伍吗丁啉对功能性消化不良血浆胃肠激素的影响[J].中国实验方剂学杂志,2015,21(13):174.