基于尿液代谢组学的附子配伍甘草减毒作用研究
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摘要
目的:利用代谢组学方法,基于附子毒性生物标记物配伍甘草前后的轨迹变化规律,阐释附子配伍甘草的减毒作用机制。方法:采用UPLC-HDMS对连续给药6个月的附子组、附子-甘草组和空白对照组大鼠尿液代谢轮廓进行分析,结合组织病理学观察结果,分析鉴定与附子毒性相关的生物标记物及相关代谢通路,并对附子与甘草配伍前后大鼠体内毒性生物标记物表达水平进行比较。结果:确定了12个与附子毒性相关的生物标记物和与之相关的6条代谢通路,附子配伍甘草后对其中10个毒性生物标记物具有较好的回调作用。结论:附子与甘草配伍可通过调节戊糖、葡萄糖醛酸转换,淀粉和蔗糖代谢,丙氨酸、天冬氨酸和谷氨酸代谢等代谢通路;有效降低附子的毒性,从代谢产物角度揭示中药配伍减毒的科学性,为附子的临床用药安全提供参考。
Objective:Using metabolomics method to explore the toxicity-attenuating mechanism of Fuzi compatibility with Gancao based on the trajectory changes of Fuzi toxic biomarkers before and after compatibility.Methods:The UPLC-HDMS was used to analyze the urine metabolic profile of the Fuzi group,Fuzi-Gancao group and control group for 6 months,and the histopathological observations were used to identify the biomarkers associated with Fuzi toxicity.The related metabolic pathways were compared and the expression levels of toxic biomarkers in rats before and after the combination of Fuzi and Gancao were also compared.Results:Twelve biomarkers related to Fuzi toxicity and 6 metabolic pathways were identified.Fuzi compatibility with Gancao had a good callback effect on 10 toxic biomarkers.Conclusion:Fuzi compatibility with Gancao can effectively reduce the toxicity of Fuzi by regulating the metabolism of pentose,glucuronic acid,metabolism of starch and sucrose,alanine,aspartic acid and glutamic acid.It reveal the science of compatibility toxicity-attenuation from the perspective of metabolites,and provide reference for the clinical safety of Fuzi.
Objective:Using metabolomics method to explore the toxicity-attenuating mechanism of Fuzi compatibility with Gancao based on the trajectory changes of Fuzi toxic biomarkers before and after compatibility.Methods:The UPLC-HDMS was used to analyze the urine metabolic profile of the Fuzi group,Fuzi-Gancao group and control group for 6 months,and the histopathological observations were used to identify the biomarkers associated with Fuzi toxicity.The related metabolic pathways were compared and the expression levels of toxic biomarkers in rats before and after the combination of Fuzi and Gancao were also compared.Results:Twelve biomarkers related to Fuzi toxicity and 6 metabolic pathways were identified.Fuzi compatibility with Gancao had a good callback effect on 10 toxic biomarkers.Conclusion:Fuzi compatibility with Gancao can effectively reduce the toxicity of Fuzi by regulating the metabolism of pentose,glucuronic acid,metabolism of starch and sucrose,alanine,aspartic acid and glutamic acid.It reveal the science of compatibility toxicity-attenuation from the perspective of metabolites,and provide reference for the clinical safety of Fuzi.
引文
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[17] TONG Q,SONG J,YANG G J,et al.Simultaneous determination of tryptophan,kynurenine,kynurenic acid,xanthurenic acid and 5-hydroxytryptamine in human plasma by LC-MS/MS and its application to acute myocardial infarction monitoring[J].Biomed Chromatogr,2018,32(4):1-7.
[18] SONG P,RAMPRASATH T,WANG H,et al.Abnormal kynurenine pathway of tryptophan catabolism in cardiovascular diseases[J].Cell Mol Life Sci,2017,74(16):2899-2916.
[19] ITO T,HANAHATA Y,KINE K,et al.Tissue Taurine Depletion Induces Profibrotic Pattern of Gene Expression and Causes Aging-Related Cardiac Fibrosis in Heart in Mice[J].Biol Pharm Bull,2018;41(10):1561-1566.
[20] SCHAFFER S,JONG C J,SHETEWY A,et al.Impaired Energy Production Contributes to Development of Failure in Taurine Deficient Heart[J].Adv Exp Med Biol,2017,975:435-446.
[21] ZHOU J T,SUN L J,CHEN L W,et al.Comprehensive metabolomic and proteomic analyses reveal candidate biomarkers and related metabolic networks in atrial fibrillation[J].Metabolomics,2019,15(7):96.
[22] SCHWINGEL P A,COTRIM H P,SALLES B R,et al.Anabolic-androgenic Steroids:A Possible Newrisk Factor of Toxicant-associated Fatty Liver Disease[J].Liver Inter-national,2011,31(3):348-353.
[23] ZHANG C E,NIU M,LI Q,et al.Urine metabolomics study on the liver injury in rats induced by raw and processed Polygonum multiflorum integrated with pattern recognition and pathways analysis[J].J Ethnopharmacol,2016,194:299-306.
[24] TAKAHASHI K,MURATA S,FUKUNAGA K,et al.Human platelets inhibit liver fibrosis in severe combined immunodeficiency mice[J].World J Gastroenterol,2013,19(32):5250-5260.
[25] JURECKA A,ZIKANOVA M,KMOCH S,et al.Adenylosuccinate lyase deficiency[J].J Inherit Metab Dis,2015,38(2):231-242.
[26] YANG Y,YIN X J,GUO H M,et al.Identification and comparative analysis of the major chemical constituents in the extracts of single fuzi herb and fuzi-gancao herb-pair by UFLC-IT-TOF/MS[J].Chin J Nat Med,2014,12(7):542-553.
[27] 章津铭,李玲,高飞,等.附子配伍甘草前后汤液沉积物的HPLC-MS化学组成分析[J].药学学报,2012,47(11):1527-1533.
[28] ZHANG J M,WAN L,HE Y X,et al.Study on intestinal absorption and pharmacokinetic characterization of diester diterpenoid alkaloids in precipitation derived from fuzi-gancao herb-pair decoction for its potential interaction mechanism investigation[J].J Ethnopharmacol,2013,147(1):128-135.
[2] SUN H,NI B,ZHANG A H,et al.Metabolomics study on Fuzi and its processed products using ultra-performance liquid-chromatography/electrospray-ionization synapt high-definition mass spectrometry coupled with pattern recognition analysis[J].Analyst,2012,137:170-185.
[3] LUO C M,YI F L,XIA Y L,et al.Comprehensive quality evaluation of the lateral root of Aconitum carmichaelii Debx.(Fuzi):Simultaneous determination of nine alkaloids and chemical fingerprinting coupled with chemometric analysis[J].J Sep Sci,2019,42(5):980-990.
[4] FU X,LU R R,ZHAO S S.Simultaneous quantitation of six aconitum alkaloids and three flavonoids in the herb couple of radix aconiti lateralis-radix glycyrrhizae(Fuzi-Gancao)by UHPLC-ESI-MS/MS[J].Pharmacogn Mag,2017,13(51):425-429.
[5] YANG Y,YIN X J,GUO H M,et al.Identification and comparative analysis of the major chemical constituents in the extracts of single fuzi herb and fuzi-gancao herb-pair by UFLC-IT-TOF/MS[J].Chin J Nat Med,2014,12(7):542-553.
[6] ZHANG J M,WAN L,HE Y X,et al.Study on intestinal absorption and pharmacokinetic characterization of diester diterpenoid alkaloids in precipitation derived from fuzi-gancao herb-pair decoction for its potential interaction mechanism investigation[J].J Ethnopharmacol,2013,147(1):128-135.
[7] LIU Y Y,WEI M Y,YUE K X,et al.Non-target metabonomic method provided new insights on the therapeutical mechanism of Gancao Fuzi decoction on rheumatoid arthritis rats[J].J Chromatogr B Analyt Technol Biomed Life Sci,2019,1105:93-103.
[8] WANG L Q,HE Y,ZHANG Y Y,et al.Effects of active components of Fuzi and Gancao compatibility on Bax,Bcl-2,and Caspase-3 in chronic heart failure rats[J/OL].Evid Based Complement Alternat Med,2016[2018-05-15]http://dx.doi.org/10.1155/2016/7686045.
[9] 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.
[10] SUN B,ZHANG M,ZHANG Q,et al.Metabonomics study of the effects of pretreatment with glycyrrhetinic acid on mesaconitine-induced toxicity in rats[J].J Ethnopharmacol,2014,154(3):839-846.
[11] HAN L F,WANG P,WANG Y L,et al.Rapid Discovery of the Potential Toxic Compounds in Polygonum multiflorum by UHPLC/Q-Orbitrap-MS-Based Metabolomics and Correlation Analysis[J].Front Pharmacol,2019,10:329.
[12] XIA T S,DONG X,LIN L Y,et al.Metabolomics profiling provides valuable insights into the underlying mechanisms of Morinda officinalis on protecting glucocorticoid-induced osteoporosis[J].J Pharm Biomed Anal,2019,166:336-346.
[13] WANG X J,WANG H Y,ZHANG A H,et al.Metabolomics Study on the Toxicity of Aconite Root and Its Processed Products Using Ultraperformance Liquid-Chromatography/Electrospray-Ionization Synapt High-Definition Mass Spectrometry Coupled with Pattern Recognition Approach and Ingenuity Pathways Analysis[J].J Proteome Res,2012,11:1284-1301.
[14] XIA F B,LI A,CHAI Y S,et al.UPLC/Q-TOFMS-Based Metabolomics Approach to Reveal the Protective Role of Other Herbs in An-Gong-Niu-Huang Wan Against the Hepatorenal Toxicity of Cinnabar and Realgar[J].Front Pharmacol,2018,9:618.
[15] WANG Y J,FENG F.Evaluation of the Hepatotoxicity of the Zhi-Zi-Hou-Po Decoction by Combining UPLC-Q-Exactive-MS-Based Metabolomics and HPLC-MS/MS-Based Geniposide Tissue Distribution[J].Molecules,2019,24(3):511-531.
[16] KOGA Y,TSURUMAKI H,AOKI-SAITO H,et al.Roles of Cyclic AMP Response Element Binding Activation in the ERK1/2 and p38 MAPK Signalling Pathway in Central Nervous System,Cardiovascular System,Osteoclast Differentiation and Mucin and Cytokine Production[J].Int J Mol Sci,2019,20(6):1346-1369.
[17] TONG Q,SONG J,YANG G J,et al.Simultaneous determination of tryptophan,kynurenine,kynurenic acid,xanthurenic acid and 5-hydroxytryptamine in human plasma by LC-MS/MS and its application to acute myocardial infarction monitoring[J].Biomed Chromatogr,2018,32(4):1-7.
[18] SONG P,RAMPRASATH T,WANG H,et al.Abnormal kynurenine pathway of tryptophan catabolism in cardiovascular diseases[J].Cell Mol Life Sci,2017,74(16):2899-2916.
[19] ITO T,HANAHATA Y,KINE K,et al.Tissue Taurine Depletion Induces Profibrotic Pattern of Gene Expression and Causes Aging-Related Cardiac Fibrosis in Heart in Mice[J].Biol Pharm Bull,2018;41(10):1561-1566.
[20] SCHAFFER S,JONG C J,SHETEWY A,et al.Impaired Energy Production Contributes to Development of Failure in Taurine Deficient Heart[J].Adv Exp Med Biol,2017,975:435-446.
[21] ZHOU J T,SUN L J,CHEN L W,et al.Comprehensive metabolomic and proteomic analyses reveal candidate biomarkers and related metabolic networks in atrial fibrillation[J].Metabolomics,2019,15(7):96.
[22] SCHWINGEL P A,COTRIM H P,SALLES B R,et al.Anabolic-androgenic Steroids:A Possible Newrisk Factor of Toxicant-associated Fatty Liver Disease[J].Liver Inter-national,2011,31(3):348-353.
[23] ZHANG C E,NIU M,LI Q,et al.Urine metabolomics study on the liver injury in rats induced by raw and processed Polygonum multiflorum integrated with pattern recognition and pathways analysis[J].J Ethnopharmacol,2016,194:299-306.
[24] TAKAHASHI K,MURATA S,FUKUNAGA K,et al.Human platelets inhibit liver fibrosis in severe combined immunodeficiency mice[J].World J Gastroenterol,2013,19(32):5250-5260.
[25] JURECKA A,ZIKANOVA M,KMOCH S,et al.Adenylosuccinate lyase deficiency[J].J Inherit Metab Dis,2015,38(2):231-242.
[26] YANG Y,YIN X J,GUO H M,et al.Identification and comparative analysis of the major chemical constituents in the extracts of single fuzi herb and fuzi-gancao herb-pair by UFLC-IT-TOF/MS[J].Chin J Nat Med,2014,12(7):542-553.
[27] 章津铭,李玲,高飞,等.附子配伍甘草前后汤液沉积物的HPLC-MS化学组成分析[J].药学学报,2012,47(11):1527-1533.
[28] ZHANG J M,WAN L,HE Y X,et al.Study on intestinal absorption and pharmacokinetic characterization of diester diterpenoid alkaloids in precipitation derived from fuzi-gancao herb-pair decoction for its potential interaction mechanism investigation[J].J Ethnopharmacol,2013,147(1):128-135.