基于代谢组学探讨芪杉方对肺癌模型小鼠的调控机制
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摘要
目的研究芪杉方干预肺癌模型小鼠内源性代谢物的变化,探索与肺癌密切相关的代谢物、代谢通路及其芪杉方的治疗作用。方法建立Lewis肺癌模型小鼠,随机分成芪杉方组、模型组和空白组。利用UPLC-HDMS技术平台对各组小鼠血液进行代谢组学分析,Ezionfo软件分析质谱代谢轮廓,通过非监督型主成分分析(OPLS-DA),结合HMDB、KEGG数据库检索和Progenesis QI软件鉴定化合物,分析肺癌相关的潜在代谢生物标记物。进一步研究芪杉方对血液代谢轮廓及生物标志物的影响,探讨芪杉方干预Lewis肺癌模型小鼠血液代谢组学的机制。结果确定22个可能与肺癌相关的代谢生物标记物。发现芪杉方对肺癌相关的18个代谢标记物具有明显的回调作用,进一步将鉴定得到的具有回调作用的标志物进行MetPA分析,得到7个代谢通路,分别是:嘧啶代谢;不饱和脂肪酸的生物合成;硒氨基酸代谢;磷酸戊糖途径;谷胱甘肽代谢;半胱氨酸和蛋氨酸代谢;甘氨酸丝氨酸苏氨酸代谢。结论基于代谢组学的研究方法发现芪杉方通过调节肺癌相关的代谢生物标记物,对肺癌起到了一定的治疗作用。
Objective To study the changes of endogenous metabolites in mouse model of lung cancer after intervention by Qishan Fang(Milkvetch Root and Taxus chinensis Formula), and investigate the metabolites and metabolic pathway related closely to lung cancer, and therapeutic effect of Qishan Fang. Methods The mouse model of Lewis lung cancer was established and all mice were randomly divided into Qishan Fang group, model group and blank group. The blood samples of all groups were given metabonomics analysis by using UPLC-HDMS technology platform. The mass spectrometry metabolic profiling was analyzed by using Ezionfo software. The potential metabolic biomarkers related to lung cancer were analyzed through OPLS-DA combined with retrieving of HMDB and KEGG and Progenesis QI software for compound identification. The influence of Qishan Fang on blood metabolic profiling and biomarkers were further studied, and mechanism of Qishan Fang in intervening blood metabonomics in Lewis mouse model of lung cancer. Results There were 22 metabolic biomarkers may related to lung cancer. It was found that Qishan Fang had significantly callback effect on 18 metabolic biomarkers related to lung cancer. The identified biomarkers with callback effects were further given MetPA analysis and 7 metabolic pathways were obtained, including pyrimidine metabolism, biosynthesis of unsaturated fatty acids, selenoamino acid metabolism, pentose phosphate pathway, glutathione metabolism, cysteine and methionine metabolism, and glycine serine and threonine metabolism. Conclusion It was found that has some therapeutic effects on lung cancer through regulating metabolic biomarkers related to lung cancer based on metabonomics research method.
Objective To study the changes of endogenous metabolites in mouse model of lung cancer after intervention by Qishan Fang(Milkvetch Root and Taxus chinensis Formula), and investigate the metabolites and metabolic pathway related closely to lung cancer, and therapeutic effect of Qishan Fang. Methods The mouse model of Lewis lung cancer was established and all mice were randomly divided into Qishan Fang group, model group and blank group. The blood samples of all groups were given metabonomics analysis by using UPLC-HDMS technology platform. The mass spectrometry metabolic profiling was analyzed by using Ezionfo software. The potential metabolic biomarkers related to lung cancer were analyzed through OPLS-DA combined with retrieving of HMDB and KEGG and Progenesis QI software for compound identification. The influence of Qishan Fang on blood metabolic profiling and biomarkers were further studied, and mechanism of Qishan Fang in intervening blood metabonomics in Lewis mouse model of lung cancer. Results There were 22 metabolic biomarkers may related to lung cancer. It was found that Qishan Fang had significantly callback effect on 18 metabolic biomarkers related to lung cancer. The identified biomarkers with callback effects were further given MetPA analysis and 7 metabolic pathways were obtained, including pyrimidine metabolism, biosynthesis of unsaturated fatty acids, selenoamino acid metabolism, pentose phosphate pathway, glutathione metabolism, cysteine and methionine metabolism, and glycine serine and threonine metabolism. Conclusion It was found that has some therapeutic effects on lung cancer through regulating metabolic biomarkers related to lung cancer based on metabonomics research method.
引文
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[16] 王亚超,张启明,王焕莉,等.中医肺藏系统的功能性症状[J].北京中医药大学学报,2017,40(4):269-271.Wang YC,Zhang QM,WANG HL,et al.Functional symptoms of the lung system in traditional Chinese medicine[J].Journal of Beijing University of Traditional Chinese Medicine,2017,40(4):269-271.
[2] Wu XF,Ruan L,Yang Y,et al.Analysis of gene expression changes associated with human carcinoma-associated fbroblasts in non-small cell lung carcinoma[J].Biol Res,2017,50(6):1-8.
[3] 秦昆明,王彬,蔡宝昌,等.代谢组学在中药现代研究的应用与展望[J].中国中药杂志,2014,39(16):3010-3017.Qin KM,Wang B,Cai BC,et al.Perspective and application of metabonomics in modern study of traditional Chinese medicine[J].China Journal of Chinese Materia Medica,2014,39(16):3010-3017.
[4] Valdes AM,Glass D,Spector TD.Omics technologies and the study of human ageing[J].Nature Review Genetic,2013,14(9):601-605.
[5] 隋雨桐,李泽光,梁华,等.四君子汤对自然衰老小鼠干预作用的代谢组学研究[J].中医药学报,2017,45(1):16-20.Sui YT,Li ZG,Liang H,et al.Metabolism of intervening effect of Sijunzi Decoction on the natural aging mice[J].Acta Chinese Medicine and Pharmacology,2017,45(1):16-20.
[6] 王喜军.药效物质基础研究的系统方法学—中医方证代谢组学[J].中国中药杂志,2015,40(1):13-17.Wang XJ.Methodology for systematic analysis of in vivo efficacy material base of traditional Chinese medicine-chinmedomics[J].China Journal of Chinese Materia Medica,2015,40(1):13-17.
[7] 隋雨桐,迟文成,姜家康,等.中医治疗非小细胞肺癌临证用药规律的研究[J].中医药学报,2017,45(4):44-47.Sui YT,Chi WC,Jiang JK,et al.Medication regularity of Chinese medicine for non-small cell lung cancer[J].Acta Chinese Medicine and Pharmacology,2017,45(4):44-47.
[8] 姜家康,迟文成,宋立佳,等.芪杉胶囊联合化疗治疗晚期非小细胞肺癌的临床疗效观察[J].中国中医基础医学杂志,2017,23(3):359-361.Jiang JK,Chi WC,Song LJ,et al.Clinical observation on effect of Qishan Capsule combined with chemotheraphy in the treatment of advanced non-small cell lung cancer (NSCLC)[J].Journal of Basic Chinese Medicine,2017,23(3):359-361.
[9] 隋雨桐,迟文成,姜家康.芪杉方联合顺铂对肺癌A549细胞PI3K/AKt信号通路的影响[J].北京中医药大学学报,2018,41(7):553-558.Sui YT,Chi WC,Jiang JK.Influence of Qishan Fang combined with cisplatin on PI3K/AKt signal pathway of lung cancer A549 cells[J].Journal of Beijing University of Traditional Chinese Medicine,2018,41(7):553-558.
[10] 隋雨桐,迟文成,姜家康.芪杉胶囊对肺癌A549细胞PI3K/AKt信号通路影响的研究[J].中医药学报,2018,46(4):22-27.Sui YT,Chi WC,Jiang JK.Effect of Qishan Capsule on PI3KAKt signaling pathway in lung cancer A549 cells[J].Acta Chinese Medicine and Pharmacology,2018,46(4):22-27.
[11] 马俊杰,王小龙,刘会平.不同证型非小细胞肺癌患者肿瘤组织代谢组学研究[J].中国中西医结合杂志,2015,35(6):659-663.Ma JJ,Wang XL,Liu HP.Metabolomics study on non-small cell lung cancer patients with different Chinese medical syndrmes[J].Chinese Journal of Integrated Traditional and Western Medicine,2015,35(6):659-663.
[12] 王均衡,李英帅,岳子敬,等.阳虚体质者粪便的代谢组学研究[J].北京中医药大学学报,2017,40(1):65-69.Wang JH,Li YS,Yue ZJ,et al.NMR-based metabonomics study on faeces of subjects with yang-deficiency constitution[J].Journal of Beijing University of Traditional Chinese Medicine,2017,40(1):65-69.
[13] 余柳莹,鲍轶,莫娟芬,等.亚甲基四氢叶酸还原酶基因多态性检测在肿瘤患者中的临床价值[J].肿瘤学杂志,2018,24(11):1103-1106.Yu LY,Bao Yi,Mo JF,et al.Detection of MTHFR gene polymorphism in malignant tumor patients[J].Journal of Chinese Oncology,2018,24(11):1103-1106.
[14] 杨辉,郭春燕.二甲砷基谷胱甘肽对肿瘤的治疗作用及机制[J].中国新药与临床杂志,2011,30(2):99-102.Yang H,Guo CY.Therapeutic effect and its mechanisms of darinaprsin on tumor[J].Chinese Journal of New Drugs and Clinical Remedies,2011,30(2):99-102.
[15] 邱红,蔡俊,邢继成,等.恶性肿瘤患者红细胞谷胱甘肽的测定及临床意义[J].临床医药实践,2005,14(9):655-656.Qiu H,Cai J,Xing JC,et al.Detection and clinical significance of erythrocyte glutathione in patients with malignant tumors [J].Proceeding of Clinical Medicine,2005,14(9):655-656.
[16] 王亚超,张启明,王焕莉,等.中医肺藏系统的功能性症状[J].北京中医药大学学报,2017,40(4):269-271.Wang YC,Zhang QM,WANG HL,et al.Functional symptoms of the lung system in traditional Chinese medicine[J].Journal of Beijing University of Traditional Chinese Medicine,2017,40(4):269-271.