基于~1H-NMR代谢组学分析山楂不同炮制品对食积症的影响
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摘要
目的:采用~1H-NMR技术的血清代谢组学方法,并结合传统生化指标分析,研究炮制对山楂促消化作用的影响。方法:实验动物喂饲特制高蛋白、高热量饲料复制食积模型,以小鼠小肠推进率和大鼠胃肠道激素水平作为药效学指标,应用~1H-NMR测定大鼠血清,结合多元统计分析方法对比给予大鼠净山楂、炒山楂、焦山楂后血清代谢物的变化,比较各炮制品对食积模型大鼠的治疗作用。结果:山楂各炮制品组对于食积因素引起的小鼠肠推进障碍及胃肠激素分泌紊乱均有改善作用,且以焦山楂组改善效果最为显著。大鼠血清代谢组学结果显示,模型组与空白组存在明显的代谢差异,根据S-plot图的变量重要性投影(VIP)值、单因素ANOVA以及受试者工作特征曲线(ROC曲线)下的面积(AUC)筛选出3-羟基丁酸,甘油磷酰胆碱(GPC),N-乙酰糖蛋白,O-乙酰糖蛋白,氧化三甲胺,丙氨酸,乙酸,谷氨酸,谷氨酰胺,肌酸,亮氨酸,乳酸,葡萄糖共13个与食积相关的生物标志物。比较给药后山楂各炮制品组引起的代谢变化,发现焦山楂组对O-乙酰糖蛋白、谷氨酰胺和GPC的回调趋势更接近空白组,其余代谢产物组成与净山楂组和炒山楂组基本一致。结论:山楂炮制后消食导滞作用存在一定差异,焦山楂改善食积疗效最为显著,可能主要通过调节胃肠动力障碍及能量代谢等发挥作用。
Objective: The effect of processing on promoting digestion of Crataegi Fructus was investigated by the method of serum metabonomics based on~1H-NMR and the analysis of traditional biochemical indexes. Method: The dyspepsia model was induced by high protein and calorie diet. The small intestine propulsive rate in mice and the hormone level in rat gastrointestinal tract were used as pharmacodynamic indexes.After administration of Crataegi Fructus,fried and charred Crataegi Fructus,the changes of serum metabolites were analyzed by~1H-NMR technique combined with multivariate statistical analysis,so as to compare the therapeutic effects of each processed products on rats with dyspepsia model. Result: Each processed products group of Crataegi Fructus could improve the ability of intestinal propulsion and gastric emptying,especially in charred Crataegi Fructus group,caused by dyspepsia. The serum metabonomics proved that there were significant metabolic differences between the model group and the blank group. A total of 13 biomarkers related to dyspepsia,covering3-hydroxybutyric acid, glycerophosphoryl choline( GPC), N-acetylglycoprotein, O-acetylglycoprotein,trimethylamine oxide( TMAO),alanine,acetic acid,glutamic acid,glutamine,creatine,leucine,lactic acid and glucose,were screened on the grounds of VIP value of S-plot,single factor ANOVA and area under receiver operating characteristic curve( ROC curve)( AUC). The metabolite composition of each administration group was the same except that the callback tendency of O-acetylglycoprotein,glutamine and GPC in charred Crataegi Fructus group was closer to that in the blank group. Conclusion: There are differences in the effect of promoting digestion to eliminate stagnation among processed products of Crataegi Fructus,charred Crataegi Fructus is more effective in improving dyspepsia,which may play a role in regulating gastrointestinal motility and energy metabolism.
Objective: The effect of processing on promoting digestion of Crataegi Fructus was investigated by the method of serum metabonomics based on~1H-NMR and the analysis of traditional biochemical indexes. Method: The dyspepsia model was induced by high protein and calorie diet. The small intestine propulsive rate in mice and the hormone level in rat gastrointestinal tract were used as pharmacodynamic indexes.After administration of Crataegi Fructus,fried and charred Crataegi Fructus,the changes of serum metabolites were analyzed by~1H-NMR technique combined with multivariate statistical analysis,so as to compare the therapeutic effects of each processed products on rats with dyspepsia model. Result: Each processed products group of Crataegi Fructus could improve the ability of intestinal propulsion and gastric emptying,especially in charred Crataegi Fructus group,caused by dyspepsia. The serum metabonomics proved that there were significant metabolic differences between the model group and the blank group. A total of 13 biomarkers related to dyspepsia,covering3-hydroxybutyric acid, glycerophosphoryl choline( GPC), N-acetylglycoprotein, O-acetylglycoprotein,trimethylamine oxide( TMAO),alanine,acetic acid,glutamic acid,glutamine,creatine,leucine,lactic acid and glucose,were screened on the grounds of VIP value of S-plot,single factor ANOVA and area under receiver operating characteristic curve( ROC curve)( AUC). The metabolite composition of each administration group was the same except that the callback tendency of O-acetylglycoprotein,glutamine and GPC in charred Crataegi Fructus group was closer to that in the blank group. Conclusion: There are differences in the effect of promoting digestion to eliminate stagnation among processed products of Crataegi Fructus,charred Crataegi Fructus is more effective in improving dyspepsia,which may play a role in regulating gastrointestinal motility and energy metabolism.
引文
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[28]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):253-255,260.
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[2]马传江,王信,辛义周,等.中药传统炮制理论的现代研究概述[J].中草药,2018,49(3):512-520.
[3]龚千峰.中药炮制学[M].北京:中国中医药出版社,2012:139-140.
[4]张三印,周艳霞,孙改侠,等.山楂不同炮制品对胃肠平滑肌收缩的影响研究[J].中药材,2009,32(10):1519-1522.
[5]张良,姜思凡,万军,等.炮制对山楂主要化学成分的影响[J].长春中医药大学学报,2014,30(1):31-34.
[6] Nicholson J K,Connelly J,Lindon J C,et al.Metabonomics:a platform for studying drug toxicity and gene function[J]. Nat Rev Drug Discov,2002,1(2):153-161.
[7]温慧萍,雷伟敏,吴宇锋,等.柳叶蜡梅对脂肪乳剂致食积证模型大鼠的影响[J].中药药理与临床,2013,29(5):82-84.
[8]刘武平,李婵艺,黄晶,等.脾气虚证大鼠尿液生物标记物的识别及其生物学意义[J].中国中药杂志,2017,42(24):4855-4863.
[9] Robin X,Turck N,Hainard A,et al. pROC:an opensource package for R and S+to analyze and compare ROC curves[J]. BMC bioinformatics,2011,doi:10. 1186/1471-2105-12-77.
[10]刘彩春,刘欢,谷陟欣,等.基于1H-NMR代谢组学的驴胶补血颗粒补血作用机制研究[J].中草药,2016,47(7):1142-1148.
[11]王丽芳,高文远,徐鑫,等.神曲鲜干品组方对食积小鼠胃肠动力及肠道菌群调整的影响[J].中国实验方剂学杂志,2017,23(4):20-24.
[12]奚锦要,朱永钦,朱永苹,等.香砂六君子汤联合针刺疗法治疗功能性消化不良临床疗效观察[J].中国实验方剂学杂志,2017,23(20):157-162.
[13]王小娟,阳松威,郭璇,等.舒胃汤对功能性消化不良大鼠小肠ICC细胞IP3R,RyR表达的影响[J].中国实验方剂学杂志,2018,24(3):97-103.
[14]徐瑶.“焦三仙”炒焦增强消食导滞的“焦香气味”物质及其协同增效作用机理研究[D].成都:西南交通大学,2018.
[15]张欣,廖祥伟,吴松,等. 5-羟甲基-2-呋喃甲醛衍生物的合成及降血糖活性评价[J].中国药物化学杂志,2012,22(5):349-355.
[16] HOU Y C,Ching H,CHAO P D,et al. Effects of glucose fructose and 5-hydroxymethyl-2-furaldehyde on the presystemic metabolism and absorption of glycyrrhizin in rabbits[J]. J Pharm Pharmacol,2005,57(2):247-251.
[17]张光辉,张进峰,周宏建.腹腔镜直肠癌根治术治疗直肠癌的疗效及对血清CRP、胃肠激素的影响[J].实用癌症杂志,2018,33(10):1607-1609.
[18]王丹,张红星,周利,等.功能性消化不良外周机制研究进展[J].湖北中医药大学学报,2018,20(5):113-116.
[19]方国兴,姚静婵,胡国华,等.四逆散加味联合布拉氏酵母菌治疗小儿功能性消化不良的临床疗效及对血清细胞因子的影响[J].中国实验方剂学杂志,2019,25(3):120-124.
[20]胡丽波,谢津璧,万坚,等.血清胃蛋白酶原及胃泌素在胃部疾病中的表达及意义[J].胃肠病学和肝病学杂志,2015,24(3):281-283.
[21]祝婧,钟凌云,叶喜德,等.枳壳不同炮制品的燥性比较及其对功能性消化不良大鼠胃肠功能的影响[J].中国实验方剂学杂志,2017,23(22):20-26.
[22] CUI J,LIU Y,HU Y,et al. NMR-based metabonomics and correlation analysis reveal potential biomarkers associated with chronic atrophic gastritis[J]. J Pharm Biomed Anal,2017,132:77-86.
[23] Fukao T,Mitchell G,Sass J O,et al. Ketone body metabolism and its defects[J]. J Inherit Metab Dis,2014,37(4):541-551.
[24] LUO L,HU M,LI Y,et al. Association between metabolic profile and microbiomic changes in rats with functional dyspepsia[J]. RSC Adv,2018,8(36):20166-20181.
[25]彭绍忠,罗燕玉,黄晓丹,等.小儿七星茶对食积模型小鼠免疫功能调节作用及其机制研究[J].中外医学研究,2018,16(32):179-181.
[26] Luong M,Lam J S,CHEN J,et al. Effects of fungal Nand O-linked mannosylation on the immunogenicity of model vaccines[J]. Vaccine,2007,25(22):4340-4344.
[27]张雪,文国容,金海,等.谷氨酰胺代谢在消化系统肿瘤中的研究进展[J].医学综述,2017,23(22):4408-4413.
[28]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):253-255,260.
[29] QIU J L,SONG B L,WANG Y J,et al. Role of glutamine in the mediation of E-cadherin,p120-catenin and inflammation in ventilator-induced lung injury[J]. Chin Med J(Engl),2018,131(7):804-812.