基于中医方证代谢组学分析四物汤对自然衰老小鼠的干预作用
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摘要
目的:通过代谢组学的研究方法观察四物汤对自然衰老小鼠体内代谢物、代谢通路的影响,分析四物汤干预自然衰老小鼠的相关靶点和作用机制。方法:选取20月龄自然哀老模型小鼠(相当于人类60～65岁)作为实验对象,同时设立3月龄小鼠作为青年组。运用UPLC-Q-TOF-MS技术对小鼠血浆进行分析,流动相含0. 1%甲酸的乙腈溶液(A)-0. 1%甲酸水溶液(B)梯度洗脱(0～15 min,2%～100%A),选择电喷雾离子源的正离子模式;采用主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)研究代谢标志物,并归纳其代谢途径。结果:四物汤对16种衰老相关代谢物的表达水平有明显回调作用,其中9种代谢标志物存在极显著统计学意义(P <0. 01);主要影响了谷胱甘肽代谢、嘧啶代谢、精氨酸和脯氨酸代谢、硒氨基酸代谢等代谢途径。结论:四物汤可影响20月龄自然衰老小鼠血浆中的代谢物,通过谷胱甘肽代谢、嘧啶代谢、精氨酸和脯氨酸代谢等途径改善小鼠衰老过程中的代谢紊乱,可为该复方后续的干预衰老物质基础研究提供生物学信息。
Objective: Through metabonomics research methods,the effect of Siwutang on metabolites and metabolic pathways in natural aging mice were observed. The related targets and mechanism of Siwutang intervention in natural aging mice were analyzed. Method: Taking 20-month-old natural aging model mice( equivalent to 60-65 years old of human beings) as the experimental subjects,at the same time,mice aged 3 months were established as the youth group. UPLC-Q-TOF-MS technique was employed to analyze the mouse plasma with mobile phase of acetonitrile( containing 0. 1% formic acid)-0. 1% formic acid solution for gradient elution and positive ion mode of electrospray ionization,and the metabolic markers were analyzed by principal component analysis( PCA) and orthogonal partial least squares-discriminant analysis( OPLS-DA), and their metabolic pathways were summarized. Result: Siwutang had obvious reversal effect on the expression levels of 16 aging-related metabolites,among which 9 metabolic markers were statistically significant( P < 0. 01),mainly affecting glutathione metabolism,pyrimidine metabolism,arginine and proline metabolism,selenium amino acid metabolism and other metabolic pathways. Conclusion: Siwutang can affect the metabolites in the plasma of 20-month-old natural aging mice,and the metabolic disorder during the aging process of mice can be improved by glutathione metabolism,pyrimidine metabolism,selenium amino acid metabolism and other pathways,and this paper can provide biological information for the study of material basis of this compound for aging.
Objective: Through metabonomics research methods,the effect of Siwutang on metabolites and metabolic pathways in natural aging mice were observed. The related targets and mechanism of Siwutang intervention in natural aging mice were analyzed. Method: Taking 20-month-old natural aging model mice( equivalent to 60-65 years old of human beings) as the experimental subjects,at the same time,mice aged 3 months were established as the youth group. UPLC-Q-TOF-MS technique was employed to analyze the mouse plasma with mobile phase of acetonitrile( containing 0. 1% formic acid)-0. 1% formic acid solution for gradient elution and positive ion mode of electrospray ionization,and the metabolic markers were analyzed by principal component analysis( PCA) and orthogonal partial least squares-discriminant analysis( OPLS-DA), and their metabolic pathways were summarized. Result: Siwutang had obvious reversal effect on the expression levels of 16 aging-related metabolites,among which 9 metabolic markers were statistically significant( P < 0. 01),mainly affecting glutathione metabolism,pyrimidine metabolism,arginine and proline metabolism,selenium amino acid metabolism and other metabolic pathways. Conclusion: Siwutang can affect the metabolites in the plasma of 20-month-old natural aging mice,and the metabolic disorder during the aging process of mice can be improved by glutathione metabolism,pyrimidine metabolism,selenium amino acid metabolism and other pathways,and this paper can provide biological information for the study of material basis of this compound for aging.
引文
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[11]吴兵平,王星,马文丽,等.鸟氨酸脱羧酶抗酶作为肿瘤治疗靶点的潜在价值[J].中国现代医学杂志,2013,23(24):62-66.
[12]隋雨桐,朱明雪,梁华.衰老相关疾病模型的研究进展[J].中国临床保健杂志,2017,20(3):317-320.
[13]马慧,尹若熙,郭敏,等.肉苁蓉多糖对D-半乳糖致衰老模型小鼠CREB表达的影响[J].中国实验方剂学杂志,2014,20(20):137-141.
[14]禹良艳,华永庆,朱敏,等.四物汤及其组成药对对大鼠卵巢颗粒细胞增殖的影响[J].中国实验方剂学杂志,2011,17(6):141-144.
[15]梁华,王燕,李琦玮,等.四物汤对增龄过程中12月龄雄性小鼠调节作用的代谢组学研究[J].中药药理与临床,2018,34(1):2-6.
[2]WANG X J,ZHANG A,SUN H. Future perspectives of Chinese medical formulae:chinmedomics as an effector[J]. OMICS,2012,16(7/8):414-421.
[3]梁华,杨丽.四物汤对老龄雌性大鼠性激素水平的影响[J].山东医药,2010,50(20):47-48.
[4]梁华,孙燕佩,朱明雪.四君子汤、四物汤对亚急性衰老模型雌性小鼠IL-2影响的实验研究[J].辽宁中医杂志,2013,40(2):193-194.
[5]SUN Y V,HU Y J. Integrative analysis of multi-omics data for discovery and functional studies of complex human diseases[J]. Adv Genet,2016,93:147-190.
[6]何瑶,王丽娟,刘婷婷,等.基于代谢组学技术分析四物汤治疗原发性痛经证的作用机制[J].中国实验方剂学杂志,2017,23(12):82-89.
[7]张亚莉,姚丛.《方剂学》《中药学》与《药征》带给中医教育的反思[J].中国实验方剂学杂志,2012,18(8):295-298.
[8]王玮玮,唐亮,周文龙,等.谷胱甘肽生物合成及代谢相关酶的研究进展[J].中国生物工程杂志,2014,34(7):89-95.
[9]王君明,崔瑛,申玲玲,等.中药致药源性肝损伤的氧化应激机制研究进展[J].中国实验方剂学杂志,2011,17(5):247-249.
[10]Ostrowski J,Wojciechowski K,Konturek S J,et al.Inhibitory effect of EGF on secretory response of rat parietal cells is associated with an induction of ODC[J]. Am J Physiol,1993,264(6Pt1):1428-1433.
[11]吴兵平,王星,马文丽,等.鸟氨酸脱羧酶抗酶作为肿瘤治疗靶点的潜在价值[J].中国现代医学杂志,2013,23(24):62-66.
[12]隋雨桐,朱明雪,梁华.衰老相关疾病模型的研究进展[J].中国临床保健杂志,2017,20(3):317-320.
[13]马慧,尹若熙,郭敏,等.肉苁蓉多糖对D-半乳糖致衰老模型小鼠CREB表达的影响[J].中国实验方剂学杂志,2014,20(20):137-141.
[14]禹良艳,华永庆,朱敏,等.四物汤及其组成药对对大鼠卵巢颗粒细胞增殖的影响[J].中国实验方剂学杂志,2011,17(6):141-144.
[15]梁华,王燕,李琦玮,等.四物汤对增龄过程中12月龄雄性小鼠调节作用的代谢组学研究[J].中药药理与临床,2018,34(1):2-6.