基于代谢组学分析补益气血方剂对自然衰老小鼠的影响
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摘要
目的:通过代谢组学的研究方法观察补气剂四君子汤、补血剂四物汤对自然衰老模型小鼠内源性代谢物的影响,分析补气剂、补血剂干预衰老的机制,比较气、血作用于模型小鼠的生物学靶点和代谢途径。方法:选取雌性自然哀老模型小鼠(20月龄)作为研究对象。经超高效液相色谱串联四极杆飞行时间高分辨率质谱法(UPLC-Q-TOF-MS)对小鼠血浆进行分析,流动相含有0.1%甲酸的乙腈溶液(A)-0.1%甲酸水溶液(B);梯度洗脱(0～15min,2%～100%A);选择主成分分析(PCA)和正交偏最小二乘-判别分析(OPLS-DA)分析代谢标志物,同时归纳其代谢途径。结果:发现四君子汤、四物汤分别影响了小鼠体内10种不同的衰老相关代谢标志物,使其表达水平向3月龄小鼠靠拢。这其中有棕榈酸、花生四烯酸、N-乙酰、D-赤藓糖-4-磷酸、硬脂酸、脱氧羟丁赖氨酸、甘草次酸、可可碱,8种代谢物完全相同,且差异有统计学意义(P<0.05)。代谢标志物主要参与了氨基糖和核苷酸糖代谢、不饱和脂肪酸的生物合成、磷酸戊糖途径、脂肪酸的生物合成、咖啡因代谢、线粒体中的脂肪酸延长、脂肪酸代谢等代谢途径。结论:补气剂四君子汤、补血剂四物汤可以通过调节小鼠的氨基糖和核苷酸糖代谢、磷酸戊糖途径、脂肪酸代谢等途径起到延缓衰老的作用,同时发现气、血存在相同的干预靶点和代谢途径,为研究传统中医理论"气血同源"提供了生物学依据。
Objective: To study the effect of qi-supplementing formula as Sijunzi Decoction and blood-supplementing formula as Siwu Decoction on endogenous metabolite of natural aging mice based on the metabolomic method, and the mechanism of the intervention of the blood-supplementing formula and blood-supplementing formula on aging was analyzed,and the biological targets and metabolic pathways of the qi and blood on the mice were compared. Methods: Female mice with natural aging model(20 months) were selected as study objects. The plasma of mice was analyzed by UPLC-Q-TOF-MS, and the mobile phase was acetonitrile solution(A)-0.1% formic acid solution(B). Gradient elution(0～15 min, 2%～100%A); principal component analysis(PCA) and orthogonal partial least squares(opls-da) were used to analyze metabolic markers and induce metabolic pathways. Results: It was found that Sijunzi Decoction and Siwu Decoction respectively affected 10 different aging related metabolic markers in mice, making their expression levels close to that of 3-month-old mice. Among them, palmitic acid,arachidonic acid, n-acetyl, d-erythrise-4-phosphate, stearic acid, deoxyhydroxybutyl lysine, glycyrrhetinic acid, and theobromine were all identical, with statistically significant differences(P<0.05). Metabolic markers are mainly involved in the metabolism of amino sugars and nucleotide sugars, the biosynthesis of unsaturated fatty acids, the pentose phosphate pathway, the biosynthesis of fatty acids, the metabolism of caffeine, the prolongation of fatty acids in mitochondria, and the metabolism of fatty acids.Conclusion: The qi-supplementing formula Sijunzi Decoction and blood-supplementing formula Siwu Decoction could play a role in delaying aging by regulating the metabolism of mouse amino sugars and nucleotide sugars, phosphate pentose and fatty acids.At the same time, the same intervention target and the same metabolic pathway were found in blood and gas, which provided biological basis for studying the TCM theory of ‘qi and blood homology'.
Objective: To study the effect of qi-supplementing formula as Sijunzi Decoction and blood-supplementing formula as Siwu Decoction on endogenous metabolite of natural aging mice based on the metabolomic method, and the mechanism of the intervention of the blood-supplementing formula and blood-supplementing formula on aging was analyzed,and the biological targets and metabolic pathways of the qi and blood on the mice were compared. Methods: Female mice with natural aging model(20 months) were selected as study objects. The plasma of mice was analyzed by UPLC-Q-TOF-MS, and the mobile phase was acetonitrile solution(A)-0.1% formic acid solution(B). Gradient elution(0～15 min, 2%～100%A); principal component analysis(PCA) and orthogonal partial least squares(opls-da) were used to analyze metabolic markers and induce metabolic pathways. Results: It was found that Sijunzi Decoction and Siwu Decoction respectively affected 10 different aging related metabolic markers in mice, making their expression levels close to that of 3-month-old mice. Among them, palmitic acid,arachidonic acid, n-acetyl, d-erythrise-4-phosphate, stearic acid, deoxyhydroxybutyl lysine, glycyrrhetinic acid, and theobromine were all identical, with statistically significant differences(P<0.05). Metabolic markers are mainly involved in the metabolism of amino sugars and nucleotide sugars, the biosynthesis of unsaturated fatty acids, the pentose phosphate pathway, the biosynthesis of fatty acids, the metabolism of caffeine, the prolongation of fatty acids in mitochondria, and the metabolism of fatty acids.Conclusion: The qi-supplementing formula Sijunzi Decoction and blood-supplementing formula Siwu Decoction could play a role in delaying aging by regulating the metabolism of mouse amino sugars and nucleotide sugars, phosphate pentose and fatty acids.At the same time, the same intervention target and the same metabolic pathway were found in blood and gas, which provided biological basis for studying the TCM theory of ‘qi and blood homology'.
引文
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[13]吴俏槿,杜冰,蔡尤林,等.α-亚麻酸的生理功能及开发研究进展.食品工业科技,2016,37(10):386-390
[14]Farahzadi R,Fathi E,Mesbah-Namin S A,et al.Anti-aging protective effect of L-carnitine as clinical agent in regenerative medicine through increasing telomerase activity and change in the hTERT promoter CpG island methylation status of adipose tissue-derived mesenchymal stem cells.Tissue Cel,2018,54:105-113
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[16]张惠敏,张超,梁明,等.《中医衰老自评量表》的编制及其信度与效度的初步评价.中华中医药杂志,2019,34(1):284-290
[2]胡齐,孙莹,宋雅芳,等.四君子汤对脾虚大鼠线粒体氧化损伤及能量代谢的影响.中华中医药学刊,2017,35(8):1972-1976
[3]雷超,张力华,苏婷.加味四君子汤对衰老大鼠的脾脏及细胞因子的影响.中华中医药杂志,2014,29(9):2941-2944
[4]唐于平,黄美艳,张彦华,等.四物汤类方与组方药材及其所含主要芳香酸体外抗氧化活性比较与量效关系研究.中国中西医结合杂志,2012,32(1):64-67
[5]刘萍,王平,陈刚,等.应用代谢组学探讨中医药复杂理论体系的研究思路和方法.中华中医药杂志,2011,26(5):993-998
[6]WangM,ChenL,LiuD,etal.Metabolomicshighlights pharmacological bioactivity and biochemical mechanism of traditional Chinese medicine.Chem Biol Interact,2017,273:133-141
[7]高子任.运用中医“气血相生”理论,使用益气补血法调节放疗后红白细胞减少症的基础研究.长春:长春中医药大学,2011
[8]袁冰.走向状态医学:精准医学开启的医学革命.中华中医药杂志,2017,32(4):1434-1448
[9]Maynard V.Olson.A behind-the-scenes story of precision medicine.Genomics Proteomics Bioinformatics,2017,15(1):3-10
[10]郑好飞,杨巧丽,刘颖.浅谈中医学思想与“精准医学”理念的相通之处.中华中医药杂志,2017,32(6):2369-2371
[11]Papaevgeniou N,Sakellari M, Jha S,et al.18α-Glycyrrhetinic acid proteasome activator decelerates aging and alzheimer’s disease progression in caenorhabditis elegans and neuronal cultures.Antioxid Redox Signal,2016,25(16):855-869
[12]Gao H, Yan P, Zhang S,et al.Chronic alpha-linolenic acid treatment alleviates age-associated neuropathology:Roles of PERK/eIF2αsignaling pathway.Brain Behav Immun,2016,57:314-325
[13]吴俏槿,杜冰,蔡尤林,等.α-亚麻酸的生理功能及开发研究进展.食品工业科技,2016,37(10):386-390
[14]Farahzadi R,Fathi E,Mesbah-Namin S A,et al.Anti-aging protective effect of L-carnitine as clinical agent in regenerative medicine through increasing telomerase activity and change in the hTERT promoter CpG island methylation status of adipose tissue-derived mesenchymal stem cells.Tissue Cel,2018,54:105-113
[15]隋雨桐,朱明雪,梁华.衰老相关疾病模型的研究进展.中国临床保健杂志,2017,20(3):317-320
[16]张惠敏,张超,梁明,等.《中医衰老自评量表》的编制及其信度与效度的初步评价.中华中医药杂志,2019,34(1):284-290