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17***significant difference model group compared with control group p<0.001; **p<0.01; *p<0.05; #signigicant difference therapy group compared with model group p<0.05.
Scores Comp[l] vs. Comp[S] colored by Sample Group
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Fig. 2 PCA score plot of rat urinary metabolites in each group rats. KB: control group; MX: model group 1; GY: therapy group; MXHF: model group 2. Numbers represent days of each experimental stage.
EmergingChinmedomicstotraditional Chinese medicine: Present and future
Ai-hua Zhang*, Hui Sun, Guangli Yan, Ping Wang, Ying Han, Xi-jun Wang*.
(National TCM Key Lab of Serum Pharmacochemistry, Key Lab of Chinmedomics, Heilongjiang University of Chinese Medicine, and Key Pharmacometabolomics Platform of Chinese Medicines, Heping Road 24, Harbin 150040, China. Tel. & Fax +86-451-82110818, email: aihuaz@yeah.net and xijunwangls@126.com)
Abstract
For World Health Organization proposed 'Health for All', we shouldpromote the usage of traditional medicine.With the development of society and people's health attention, TCM is getting more and more popular in the whole world for improving health condition of human beings and preventing or healing diseases. However, TCM is also facing severe challenges or problemes. The largest obstacle suffers from insufficient modern scientific research, that not only lowering the position of TCM also restricting the development of TCM in the abroad. Therefore, it is indispensable to strengthen TCM research, which undoubtedly demands significant amount of analytical power and efforts. Through system studies, we have establishedand advanced the concept of Chinmedomics that is suitable for effective evaluationandbasic research of prescription and practice characteristics of TCM, may beneficially provide an opportunity to scientifically express the meaning of the syndrome and medical formulae of TCM. Chinmedomics apporoaches as pillars of the potential bridge between chinese and western medicine, may beneficially influence and provide an opportunity to explain the true meaning of evidence-based chinese medicine. TCM can
be in a better position to be accepted by modern society, serving for human health, especiallyin combination therapies and personalized medicine etc.
Keywords:
Chinmedomics; Metabolomics; biomarkers; metabolites; traditional Chinese medicine
Introduction
Traditional Chinese Medicine (TCM) has been practiced for thousands of years in Asian countries. To enhance therapeutic effects of TCM, practitioners advocated combinatorial therapeutic strategies and often prescribe a combination of plant species and/or minerals called formulae [1]. Typically, formulae consist of several kinds of crude drugs that originate from medicinal plants, animals or minerals; one represents the principal component and is called the monarch drug in TCM, whereas the others serve as adjuvant components that facilitate the delivery of the principal component to the disease site within the body [2,3]. In formulae, the herbs work together harmoniously to achieve an ideal therapeutic outcome. It is believed that multiple herbs and constituents in the formulae could hit multiple targets and exert synergistic therapeutic efficacies. Different from single component of western drug, formulae is a complex system consisting of multiple compounds, which makes scientific evaluation of TCM more difficult [4]. Therefore, it is indispensable to strengthen formulae research, which undoubtedly demands significant amount of analytical power and efforts.
Serum pharmacochemistry of TCM is essential approach to ensure optimal drug efficacy and assess theapeutic treatment; the advances in metabolomics provide new tools to the understanding of formulae action and mechanism [5]. As a systemic approach, metabolomics adopts a 'top-down' strategy to reflect the function of organisms from terminal symptoms of metabolic network and understand metabolic changes of a complete system caused by interventions in holistic context [6]. This property is in concert with the holistic efficacy of TCM; metabolomics has the potential to impact our understanding of the theory behind the evidence-based Chinese medicine [7]. This project integrates metabolomics with serum pharmacochemistry approach of TCM, elucidating the therapeutic effect, synergistic properties and metabolism of formulae, based on the metabolic profiling, fingerprinting, metabolic markers and pathways of the animal model of TCM syndrome that has essential biological characterization, further reveal the efficacy of the material basis of the prescription in order to clarify the law of formulae compatibility. Our research will carry out the key and scientific issues in TCM including prescription design, clinical usage and drug discovery from classic formulae.
Metabolomics becomes practically available and resembles TCM in many aspects and can serve as a key driving force for translation of the formulae into practice. It has been used to investigative the metabolic profile and biomarkers, key metabolic enzymes and metabolic pathways of the jaundice syndrome (JS) and its sub-types of YangHuang (acute) and YinHuang (chronic) in patients with liver disease, as well as metabolic characteristics of liver injury animal models [8]. The global effects and possible mechanisms of Yin-Chen-Hao-Tang (YCHT) against YangHuang and Yin-Chen-Sini-Tang against YinHuang have been comprehensively explored[9].In vivo ingredients after administration of prescription were characterized in the serum, as well as its relationship with endogenous metabolic markers, to illustrate andclarify the potential basis of the relevant prescription and law of formulae compatibility. Through a large number of studies, we have establishedand advanced the concept of Chinmedomics [10]. Some characteristic examples are presented to highlight the application of this platform to research and development of TCM formulaeas well as some of the necessary milestones for moving TCM into mainstream health care.
2. Establishmentof Chinmedomics
Chinmedomicsenables study of living systems from a holistic perspective based on the profiling of multitude biochemical components, opens up a unique and novel opportunity to reinvestigate over-all symptoms and effective evaluation of formulae, and has developed in recent years from a technology-driven enterprise to a new strategic tool in TCM sciences and enhance the scientific value of the TCM and international community's awareness of Chinese
medicine.Chinmedomics, namely as elucidating the therapeutic, and synergistic properties of formula and related metabolic pathways using modern analytical techniques at small molecule metabolite level, explore the nature and essence of TCM syndromes and in vivo effective substance of prescription as well as its relationship with metabolic markers, simultaneous to clarify the law of prescription compatibility[10]. It has recently demonstrated significant potential in TCM field. Chinmedomics approach is considered to have the potential to revolutionize TCM research and to advance the development of formulae[11]. Adoption of chinmedomics approach would do much help for exploring the scientific connotation of TCM syndrome and the function of formulae, provides easily measured surrogate biomarkers, abridging TCM with molecular pharmacology. Chinmedomics holds the promise of a comprehensive, non-invasive analysis of metabolic biomarkers that could help to monitor treatment response of formulae and elucidate the therapeutic and synergistic properties of formulae, which would become a platform for the development of new therapies or drugs.
3. Application of Chinmedomics
Particularly, metabolomic study of liver injury model induced by alcohol and the intervention effects of YCHT, a classic traditional Chinese medicine formulae for treatment of JS and liver disorders in China. The greatest difference in metabolic profiling was observed from alcohol-treated rats compared with the control and YCHT-treated rats[9]. The positive ion ceramide (d18:1/25:0) was elevated in urine of alcohol-treated rats, providing further support for an involvement of the sphingomyelin signaling pathway in alcohol liver injury and the intervention effects of YCHT. JS related metabolites play an essential role in glutamate metabolism, synthesis and degradation of ketone bodies, alanine and aspartate metabolism, which are tightly correlated with the genes of neurotransmitters, hormones and cytokines in the metabolites interaction network[8]. Interestingly, 44 distinct metabolites identified from these pathways, many are in various stages of progress at the JS. Further study of these metabolites may facilitate the development of non-invasive biomarkers and more efficient therapeutic strategies for JS. Furthermore, vitamin B6 metabolism, tryptophan metabolism, arginine and proline metabolism was also the top function for YAH patient. Additionally, steroid hormone biosynthesis, primary bile acid biosynthesis, cysteine and methionine metabolism were all related with YIH. It suggested that Chinmedomics method would be helpful to establish a suitable model for reasonably evaluating disease syndrome, exploring pathological mechanism of the syndrome, clarifying the relationships between the syndrome and related diseases, also has the potential to be developed into a clinically useful diagnostic tool, and could also contribute to a further understanding of disease mechanisms. Metabolite profiling of liver-stagnation and spleen-deficiency syndrome (LSS)-type disease was performed by Chinmedomics, and more importantly, of the 12 differential metabolites, 4 metabolite markers were effective for the diagnosis of human LSS[12].
Based on biomarkers and metabolic pathways of human JS in patients, screening multiple impacting and preparation factors, the combination of the EtOH, ANIT and rhubarb extract prepared YIH rat model [13]. 19 biomarkers of JS animal model were identified. Of note, Yin Chen Si Ni Tang has a potential pharmacological effect on YIH syndrome through regulating multiple perturbed pathways to their normal state.On the basis of metabolomic characteristics of the animal model, UPLC-Q-TOF/HDMS combined with pattern recognition approaches were integrated to discover and identify in vivo component of YCHT and its compatibility, and relationship of the component associated with metabolic profiling and biomarker[14]. UPLC/MS fingerprints of the samples were firstly established in vitro and in vivo, with 45 compounds in YCHT and 21 compounds in rat plasma after oral administration of YCHT were detected. Of the 45 detected compounds in vitro, 30 were identified, and all of the 21 compounds detected in rat plasma were identified. Of the identified 21 compounds in rat plasma, 19 were the original form of compounds absorbed from the 45 detected compounds in vitro, 2 were the metabolites of the compounds existed in YCHT. 8 compounds that has good choleretic and hepatoprotective effect, can be only absorbed under the synergistic or compatibility conditions. The correlation of PK-PD of 3 main component in YCHT was simultaneously analyzed[15].The three principal components of YCHT are Artemisia
annua L., Gardenia jasminoids Ellis, and Rheum Palmatum L., whose major active ingredients are 6,7-dimethylesculetin (D), geniposide (G) and rhein (R), respectively. To determine the mechanisms that underlie this formula, we conducted a systematic analysis of the therapeutic effects of the DGR compound using chinmedomics [16].DGR synergistically causes intensified dynamic changes in metabolic biomarkers, regulates molecular networks through target proteins, has a synergistic/additive effect and activates both intrinsic and extrinsic pathways.DGR combination exerts a more robust therapeutic effect than any one or two of the three individual compounds by hitting multiple targets in a rat model of hepatic injurysyndrome.
Biological nature of kidney deficiency syndrome, heat syndromes, liver depression and spleen deficiency syndrome, insomnia, heart-qi deficiency syndrome, diabetes etc, andin vivo substance and treatment mechanisms of the related prescription had been clarified by chinmedomics[17]. Liu Wei Di Huang Wan (LW), a well-known formulae for invigorating kidney-yin, consisting of Radix Rehmanniae Preparata, Fructus Macrocarpii, Rhizoma Dioscoreae Oppositae, Poria, Rhizoma Alismatis and Cortex Moutan Radicis. The first three crude drugs are known as "Sanbu" (SB), which invigorate yin of the kidney, and the rest as "Sanxie" (SX), which attenuate the effects of invigoration. Kidney yin deficiency is a common disease in China, especially for old people. However, with the increasing pace of life, many young people begin to suffer from this disease. Therefore, the metabolic profiling of rats with kidney yin deficiency and the therapeutic effects of LW, SB and SX was to mine the differentiating metabolites of this disease for clinic diagnosis and to explore the therapeutic mechanism of LW on the global level of metabolic and signaling pathways. Targeting the metabolic pathways that contribute to the kidney function improvement of LW revealed that SB and SX partially contribute to the effects of LW on metabolism related to SF. The contribution of SB and SX was evaluated by metabolic analysis; SB was clearly superior to SX. Thus, SBis the principal ingredient in LW and SX is only an auxiliary, correctant ingredient. Increased free N-acetyl-neuraminic acid (NeuAc) results in sialic acid storage diseases that are very similar to that of Children Five Late Syndrome (FLS). The therapeutic mechanism of LW in the treatment of FLS are primarily attributed the prevention of NeuAc production by affecting NeuAc metabolism. A total of 38 compounds in LW, of the 38 detected compounds in vitro, 17 compounds were identified from SB, and 21 were identified from SX. 11 compounds in rat plasma after oral administration of LW were identified, has good tonifying kidney effect. Especially, 5-HMFA was metabolized from different drugs (Radix Rehmanniae Preparata, Rhizoma Dioscoreae Oppositae and Rhizoma Alismatis) at different times and metabolism plateau up to 18 hours to maintain concentration in the blood, and showed good anti-aging and improving blood stream. Relationship of component composition and effect has also been researched to reveal the scientific value of prescription compatibility. Chinmedomics approach may offer a powerful technique for essence research in Chinese medicine symptom, providing an experimental foundation for the clinical evaluation of kidney yin deficiency and a novel approach for the study of TCM formulae.
Globally metabolic characters of the insomnia and the therapeutic effects of Suanzaoren decoction (SZRD), mechanisms of Jujuboside A and B protects against insomnia had been explored[18]. SZRD, Jujuboside A and B administration could provide satisfactory effects on insomnia through partially regulating the perturbed pathway[19,20]. Metabolic profiling and biomarkers of insomnia, metabolomic feature network and key pathways of SZRD, Jujuboside A and B protect against insomnia have constructed to help provide insights into drug action mechanisms, and enable us to increase research productivity toward metabolomic drug discovery. Constructing metabolism network and key pathways ofLSS was to help provide insights into disease mechanisms, and increase research productivity toward chinmedomics drug discovery from formulae[12]. It demonstrates that robust chinmedomics has the potential as a non-invasive strategy and promising screening tool to evaluate the potential of these metabolites in the early diagnosis of LSS patients and provides new insight into pathophysiological mechanisms.
4. Conclusion and future perspectives
On the basis of a large number of TCM research, Chinmedomics has been put forward, mainly according to the study of pharmaco-metabolomics of Chinese medical formulae[10]. Under
the guidance of the TCM theory, the chinmedomicsapproach can be used to evaluate clinical syndromes and identify potential biomarkers. The integral chinmedomics is the best to fit the holistic concept of multi-targets and systems of TCM theory, therefore it may be one of the best methods to study the TCM science. Widespread use of this technique would significantly advance the field of TCM by bridging the gap between Chinese and Western medicine. Expect this rigorously approach is a potential tool to explore and clarify the therapeutic action of TCM. It is conceivable that the application of chinmedomic platforms will eventually lead to the reconciliation and integration TCM with contemporary medicine and systems medicine. In summary, integration of chinmedomics-based diagnostic principles into the TCM would make it possible to interpret and explore the essence of personalized TCM, and might be the direction to enable a revolution for future health care, also perhaps it is time to embrace the arrival of 'TCM-OMICS' era in Chinese medicine research.
Acknowledgments
This work was supported by grants from the Key Program of the Natural Science Foundation of the State (Grant No. 90709019), the National Key Program on the Subject of Drug Innovation (Grant No. 2009ZX09502-005), the National Specific Program on the Subject of Public Welfare (Grant No. 200807014), and the National Program for Key Basic Research Projects in China (Grant No. 2005CB523406). References
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Mechanism of Sini Freeze-Dried Powder to Improve Sleep Based on Gene
Expression in Drosophila Brain
Bian Hongsheng, Huang Lili, Jin Yang and Li Tingli
HeilongjiangUniversityofChineseMedicine,Harbin150040,China
Abstracts
Objective: By using gene chip technology, we observed the gene expression in Drosophilabrain through sleep deprivation caused by light stimulation and the intervention of Sini freeze-dried powder, in order to discuss the mechanism of Sini freeze-dried powder to improve sleep. Methods: We collect Wild-type (Canton S) female Drosophila melanogaster within 12 h of emergence without mating. After shallow anesthesia with CO2, flies are randomly divided into three groups (Control group, Sleep deprivation group and Administration group).We put the different groups of flies into glass tubes with basal medium, Continuous culture 3 days.In the beginning 4 days, we put administration group flies into the4%(Weight Ratio between Sini freeze-dried powder and basal medium) Sini freeze-dried powder medium. In the beginning 7 days, at 7:00, we put sleep
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