基于代谢组学技术研究艾灸“足三里”对慢性萎缩性胃炎大鼠胃组织代谢物的影响
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摘要
目的:采用代谢组学的方法观察正常及疾病状态下,艾灸"足三里"对胃组织代谢产物的影响,从代谢组学角度探讨阳明经穴位与胃之间相对特异性联系的基础。方法:将56只SD大鼠分为正常组、正常+艾灸组、模型组、模型+艾灸组,每组14只。通过N-甲基-N′-硝基-N-亚硝基胍联合乙醇灌胃、饥饱失常法复制慢性萎缩性胃炎(CAG)大鼠模型。正常组和模型组不予以治疗,正常组+艾灸组和模型组+艾灸组予以艾灸"足三里""中脘"穴治疗,每次15min,每日1次,共2周。HE染色观察大鼠胃组织病理变化,核磁共振氢谱(1 H NMR)技术检测胃组织代谢物并用Chenomx NMR Suite软件分析所得数据。结果:模型组大鼠胃组织病理改变符合CAG病理诊断标准,模型+艾灸组大鼠胃组织病理改变较模型组有一定程度的改善。与正常组比较,模型组大鼠胃组织中的乳酸、谷胱甘肽、N-乙酰天门冬氨酸、尿苷二磷酸葡萄糖、肌糖升高(P<0.05),亮氨酸、甜菜碱、磷酸胆碱、尿嘧啶降低(P<0.05);艾灸"足三里"后浓度发生逆转的代谢物有谷胱甘肽、N-乙酰天门冬氨酸、磷酸胆碱、尿嘧啶(和模型组比较,P<0.05)。与正常组比较,正常+艾灸组胃组织中的肌醇升高(P<0.05),而3-羟基丁酸、丝氨酸、尿苷二磷酸葡萄糖降低(P<0.05)。结论:从代谢组学角度证明了胃经穴位与胃之间存在一定的相对特异性联系,谷胱甘肽、N-乙酰天门冬氨酸、磷酸胆碱、尿嘧啶等差异性代谢物可能是介导艾灸胃经穴位产生效应的代谢物质基础。
Objective To observe the effect of moxibustion at"Zusanli"(ST36)on metabolites of gastric tissue in chronic atrophic gastritis(CAG)rats,so as to explore the correlation between Yangming Meridian and stomach based on metabolomics.Methods Fifty-six SD rats were randomly divided into normal,normal+ moxibustion,model and model+ moxibustion groups(n=14 rats/group).The CAG model was established by gavage of N-methyl-N′-nitro-N-nitroso-guanidine(MNNG)solution and irregular diet for 12 weeks.Moxibustion was applied to ST36 and"Zhongwan"(CV12)for 15 min,once daily for two weeks in the normal+ moxibustion group and model+moxibustion group.The histopathological changes of the gastric tissue were observed by H.E.staining.1 H nuclear magnetic resonance(1 H NMR)spectroscopy was used to detect gastric tissue metabolites and the data were analyzed using the Chenomx NMR Suite software.Results After modeling,the regional gastric tissue showed pale thinner gastric wall,and reduction of regional glands with necrosis and disordered arrangement,epithelial cell exfoliation,lymphocyte infiltration as well as scattered punctate hemorrhage,which was evidently milder in the model+ moxibustion group.The metabolites of lactic acid,glutathione,N-acetylaspartate,uridine diphosphate glucose and muscle sugar in the gastric tissue in the model group were increased compared with those in the normal group(P<0.05),and the metabolites of leucine,betaine,phosphorylcholine,and uracil were decreased(P<0.05).The glutathione,N-acetylaspartate,phosphorylcholine and uracil in the model+moxibustion group were significantly reversed relevant to the model group(P<0.05).The inositol in gastric tissue were significantly increased(P<0.05),3-hydroxybutyric acid,serine and uridine diphosphate glucose were significantly decreased(P<0.05)in the normal+ moxibustion group than in the normal group.Conclusion Moxibustion has a good efficacy in regulating some metabolic substances such as glutathione,N-acetylaspartic acid,choline phosphate and uracil of CAG rats,suggesting a specific relationship between the Stomach Meridian and the stomach.
Objective To observe the effect of moxibustion at"Zusanli"(ST36)on metabolites of gastric tissue in chronic atrophic gastritis(CAG)rats,so as to explore the correlation between Yangming Meridian and stomach based on metabolomics.Methods Fifty-six SD rats were randomly divided into normal,normal+ moxibustion,model and model+ moxibustion groups(n=14 rats/group).The CAG model was established by gavage of N-methyl-N′-nitro-N-nitroso-guanidine(MNNG)solution and irregular diet for 12 weeks.Moxibustion was applied to ST36 and"Zhongwan"(CV12)for 15 min,once daily for two weeks in the normal+ moxibustion group and model+moxibustion group.The histopathological changes of the gastric tissue were observed by H.E.staining.1 H nuclear magnetic resonance(1 H NMR)spectroscopy was used to detect gastric tissue metabolites and the data were analyzed using the Chenomx NMR Suite software.Results After modeling,the regional gastric tissue showed pale thinner gastric wall,and reduction of regional glands with necrosis and disordered arrangement,epithelial cell exfoliation,lymphocyte infiltration as well as scattered punctate hemorrhage,which was evidently milder in the model+ moxibustion group.The metabolites of lactic acid,glutathione,N-acetylaspartate,uridine diphosphate glucose and muscle sugar in the gastric tissue in the model group were increased compared with those in the normal group(P<0.05),and the metabolites of leucine,betaine,phosphorylcholine,and uracil were decreased(P<0.05).The glutathione,N-acetylaspartate,phosphorylcholine and uracil in the model+moxibustion group were significantly reversed relevant to the model group(P<0.05).The inositol in gastric tissue were significantly increased(P<0.05),3-hydroxybutyric acid,serine and uridine diphosphate glucose were significantly decreased(P<0.05)in the normal+ moxibustion group than in the normal group.Conclusion Moxibustion has a good efficacy in regulating some metabolic substances such as glutathione,N-acetylaspartic acid,choline phosphate and uracil of CAG rats,suggesting a specific relationship between the Stomach Meridian and the stomach.
引文
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[2]周逸平.关于经络经脉脏腑相关研究的深入探讨[J].针刺研究,2006,31(6):327-328.
[3]周逸平.经脉-脏腑相关是经络理论的核心[J].针刺研究,1999,24(3):238-241.
[4]房静远,刘文忠,李兆申,等.中国慢性胃炎共识意见[J].中华消化杂志,2007,18(3):674-684.
[5]路敏敏,金世禄,刘宝珍,等.慢性萎缩性胃炎的诊断进展[J].中华全科医学,2015,13(5):823-825.
[6]周利,何权,辛玉,等.浅谈“肚腹三里留”[J].中国针灸,2015,35(7):723-726.
[7]常小荣.严洁,易受乡,等.足三里梁门胃俞穴红外辐射与慢性胃痛的相关规律及其应用[J].世界化人消化杂志,2009,17(5):516-520.
[8]陈松,严洁,朱文锋,等.胃俞与足三里穴区红外辐射检测应用于慢性胃痛辨证的初步研究[J].湖南中医药大学学报,2001,21(1):52-53.
[9]高骏,刘旭光,颜贤忠,等.代谢组学研究针灸关键问题的初步策略分析与探讨[J].针刺研究,2011,36(4):296-301.
[10]唐雅妮,谭成富,刘薇薇,等.基于核磁共振氢谱技术研究电针“内关”穴对心肌缺血再灌注损伤大鼠血清及心肌组织代谢物的影响[J].针刺研究,2018,43(3):152-162.
[11] LAO Y M,JIANG J G,YAN L.Application of metabonomic analytical techniques in the modernization and toxicology research of traditional Chinese medicine[J].Br J Pharmacol,2009,157(7):1128-1141.
[12] ZHAO L,NICHOLSON J K,LU A,et al.Targeting the human genome-microbiome axis for drug discovery:inspirations from global systems biology and traditional Chinese medicine[J].J Proteome Res,2012,11(7):3509-3519.
[13]刘运辉.3-羟基丁酸及其衍生物影响神经细胞生理功能及合成荧光探针对细胞内吞的研究[D].汕头:汕头大学,2011.
[14]孙立业,毛璇,张冬娟,等.Ⅱ型糖尿病肾病模型db/db小鼠血浆的代谢组学研究[J].中国医药导报,2012,9(13):19-21.
[15] XU J,ZHENG X,CHENG K K,et al.NMR-based metabolomics reveals alterations of electro-acupuncture stimulations on chronic atrophic gastritis rats[J].Sci Rep,2017,7:45580.
[16]杜小正,袁博,张星华,等.基于代谢组学研究热补针法治疗类风湿关节炎寒证家兔的作用机制[J].针刺研究,2017,42(3):202-208.
[17] KUIPERS E J,UYTERLINDE A M,PEA A S,et al.Long-term sequelae of helicobacter pylori gastritis[J].Lancet,1995,345(8964):1525-1528.
[18]秦江波.正常成人不同年龄组海马氢质子磁共振波谱测定[D].太原:山西医科大学,2009.