| Wei Qi theory originated from the Huangdi neijing, is one of the basic theories of traditional Chinese medicine. " The generation, operation and function of Wei qiwere discussed in detail in Huangdi neijing. As a part of the body Yang, Wei qi plays its roles in warming yang organs, filling embellish skin, defensing external evils, which is closely related with the theory of “preventive treatment of disease”. So, the theory of Wei qi is always an important research content of TCM scientific research field. The source, operation, function and clinical application of Wei qi are paid much more attention. In recent years, with the development of modern medical research technology, some scholars attempted to deeply study the theory of Wei qi by means of modern medical technology, and put forward a series of theories about the nature of the Wei qi: including the immunity theory, thermal metabolic theory of Wei Qi, Wei Qi, neuroendocrine immune theory. Although to certain extent, the researchers summarized the conclusions and promoted the theory of weiqi, but the current research still only focuses on the nature of the search to a single function of Wei qi, while ignoring the operation method of characteristics from the Wei qi, the physical pretty rhythm and explore its disease lubricating material nature, so it is difficult to interpret the essence of Wei qi; secondly, the research about Wei qi mainly involve such as immune organs, immune cells and other aspects. These results to some extent, enriched and developed the theory of Wei qi, however, from the main function of Wei qi and the concept of holism, apparently material basis of other functions have not been involved and is not deep enough. in an overall and comprehensive level of material basis for further study of Wei qi. Metabonomics(metabolomics/metabonomics) provided the possibility, which is a new branch of genomics, transcriptomics and proteomics of systems biology. It is the system changes of pathophysiology of stimulation of metabolites, to reveal the essence of the scientific life activities of the human body metabolism, is an important part of systems biology. Group index analysis based on high-throughput detection, and data processing methods, qualitative and quantitative analysis of endogenous metabolites in biological organisms, analysis of differences in the condition of different metabolic fingerprint are used to obtain the corresponding biological markers, which reveals the overall functional state of organisms in a specific time, environment. Metabonomics puts aside the complex biochemical molecules in vivo interaction and network metabolism changes, the metabolic end product diversification comprehensive analysis, shows the change state of internal to the organism as a whole, avoiding using a single indicator or a few refers to the one sidedness of physiological and pathological changes of a certain standard, the overall concept and the traditional Chinese medicine treatment of disease and medication is consistent.The clock genes are widespread in the field of biology, its role is to generate and control biological circadian rhythm. Circadian variation widely exists in various organisms, biological, biochemical, physiological and behavior in the process have shown some characteristics of circadian rhythm. The biological clock gene can discovery and reveal the law of its activities, the study of circadian rhythm of breakthrough. While the rhythm of the circadian clock related health gas has not yet been reported. Year rhythm variation characteristics of the test time by using modern biological theory and research methods to explore the Wei qi, to provide theoretical reference and experimental basis for the clinical treatment of qi related diseases. In mammals, the regulation of biological Zhong Jiyin’s inner biological rhythm mainly include Clock, Bmall, Per, Cry, and protein composition of the feedback loop.This paper follows the overall concept and the theory of "Qi" meaning, based on the periodic variation of the annual rhythm of Wei Qi, on the basis of previous experiments, based on the nature of the year cycle, celebrated on the vernal equinox, autumnal equinox and winter solstice summer solstice, the four time points for generation of metabolomics detection on rat thymus, and the blood lymphocytes, thymocytes of biological clock gene expression determination, gene regulation mechanism of circadian rhythm changes so as to explore the Wei Qi years, looking for small molecules based reflects the annual change of Qi, that cycle in years by nature, linkage Wei Qi small molecule biological clock related genes and play defense the mechanism of the regulation mechanism. Annual cycle by studying the changes of Qi and regulating mechanism, explore the essence of Qi and material basis, prevention and treatment for clinical health Qi related diseases to provide experimental support.Part 1 Theoretical study of Wei qiObjective: to explore regular rhythm changes and related material basis from the literature of Wei qi.Methods: according to the ancient records, from the source of generation, Wei qi, circulation, function, essence, Qi, Wei qi theory of rhythm related clinical application point of view, the Wei qi are discussed in detail.Conclusion: the method of modern research of Wei Qi only focus on exploring the nature of the single function of Wei qi, ignoring the method to explore the material operation characteristics from the Wei qi, the circadian rhythm and the essence of Chinese lubricating disease, so it is difficult to interpret the Wei qi, the material basis of Weiqi is range the lack of overall and comprehensive understanding and research, metabolomics, biological clock gene technology offers the possibility of research in this area.Part 2 Experimental study of Wei qiExperiment 1 Wei qi year rhythm of thymus metabolism Based on the experimental study of a normal male SD rats ofObjective: through the method of metabonomics, to find the materials that influence the year rhythm in thymus of SD rats in the four seasons.Methods:By use of fatigue and alternately in preparation of SD male rats model of Wei qi. The normal group was given routine daily feeding, fed with isothermal pure water. In the Autumnal Equinox Day(september 23, 2013), the winter solstice(December 22, 2013), spring equinox(March 21, 2014), summer solstice(June 21, 2014) about 12:00, on the solstices and equinoxes in four groups of normal SD rats(n=20) broken blood by liquid chromatography-mass spectrometry. The determination of the metabolites of the thymus is analyzed by statistical analysis using principal component.Results: the changes in spring and summer, autumn and winter four seasons in the normal group and model group had obvious difference in the metabolic spectrum, this study in the positive ion mode has 5 main components, the accumulation of R2X=0.813, as shown in Table1.1. We conclude that c AMP(cyclic adenosine monophosphate), malic acidn(malic acid), Homocarnosine(Gao Jitai), Uridine(Niao Gan), Cortisol(cortisol) as potential markers reflect changes in Qi in different seasons.Conclusion: that c AMP(cyclic AMP), malic acidn(malic acid), Homocarnosine(Gao Jitai), Uridine(Niao Gan), Cortisol(cortisol) may represent a potential biomarker of Qi in different seasons.Experiment 2 Based on Metabonomics the study on metabolic changes of metabolites in SD rats with Wei qi deficient syndrome and Yupingfeng powder interventionObjective: Through the method of metabonomics of thymus SD on rat models of the four seasons of the material by metabonomics determination, thymus model group and treatment group on each animal material determination, and compared with the normal group, to find the same rhythm of matter and its changes with the normal group, which required Yuping powder the treatment effect.Methods: By use of fatigue and alternately in preparation of SD male rats model of Wei qi.The treatment group was given orally solution of Yuping Yupingfeng powder and the normal group was given routine daily feeding, fed with isothermal pure water.On the equinox(March 30, 2013), the winter solstice(December 22, 2013),(March 21, 2014) the spring equinox, summer solstice(June 21, 2014) about 12:00, the equinox,winter solstice, vernal equinox, summer solstice,four groups of rats(n=30) brokenblood, using liquid chromatography-mass spectrometry, for detection of metabolites serum. The analysis method for statistical analysis is principal components, and compared with the normal group, the treatment effect disproof of Yuping powder.Results:The changes of spring, summer, autumn, winter four seasons in the normal group and the treatment group with different metabolic profiles.Conclusion: Yuping Feng San to Wei Qi state correction in rats.Experiment 3 Experimental study on the expression of clock gene Clock、Bmall and Cry1 on Wei qi deficient syndrome SD rats and year rhythm intervened by Yupingfeng powderMethods: after the vernal equinox, summer solstice, winter solstice equinox, 2 weeks before the 20 SD rats were randomly divided into normal control group and model group, normal control group rats, the rats in the model group with combined cold and heat fatigue alternating method to make rat model of qi deficiency syndrome. To detect the expression of each season of rat peripheral blood lymphocyte clock gene Clock by RT-PCR method.Results:1 compared with the normal control group, model group, expression of seasonal clock gene Clock decreased, the difference was statistically significant(P<0.05). Comparison between each season: compared with the spring group, normal control group, model group, equinox, summer solstice winter solstice clock gene expression of Clock decreased, the difference was statistically significant(P<0.05); compared with the summer solstice group, normal control group, model group the clock gene Clock expression decreased, the difference was statistically significant(P<0.05); in minutes the lower expression of Clock genes had no significant difference(P>0.05); compared with autumn group, normal control group, model group the clock gene Clock expression decreased, the difference was statistically significant(P<0.05).2 Compared with normal control group, model group, expression of seasonal clock gene Bmall decreased, the difference was statistically significant(P<0.05). Comparison between each season: compared with the spring group, normal control group, model group, equinox, summer solstice winter solstice clock gene expression of Clock decreased, the difference was statistically significant(P<0.05); compared with the summer solstice group, normal control group, model group the clock gene Bmall expression decreased, the difference was statistically significant(P<0.05); in minutes the lower expression of Clock genes had no significant difference(P>0.05); compared with autumn group, normal control group, model group the clock gene Bmall expression decreased, the difference was statistically significant(P<0.05).3 compared with normal control group, model group, expression of seasonal clock gene Cry1 decreased, the difference was statistically significant(P<0.05).(2) comparison between each season: compared with the spring group, normal control group, model group, the summer solstice autumnal equinox, winter solstice clock gene Clock expression decreased, the difference was statistically significant(P<0.05); compared with the summer solstice group, normal control group, model group the clock gene Cry1 expression decreased, the difference was statistically significant(P<0.05); in minutes the lower expression of Clock genes had no significant difference(P>0.05); compared with AE group, normal control group, model group the clock gene Cry1 expression decreased, the difference was statistically significant(P<0.05).Conclusions:1 There are seasonal changes of the expression of lymphocyte clock gene in Wei qi deficiency model rats and normal rats, and the changes in these two group are related. The expression of Clock increased by the intervention of Yupingfeng powder(P<0.05).2 there are seasonal changes of the expression of Wei Qi deficiency model rats and normal rats blood clock gene Clock, and the changes in these two group are related. The expression of Bmal1 increased in spring and winter by intervened with Yupingfeng powder(P<0.05)The expression of Bmal1 increased in summer and autumn by intervened with Yupingfeng powder(P>0.05).3 There are seasonal changes of the expression of lymphocyte Cry1 gene in Wei qi deficiency model rats and normal rats, and the changes in these two group are related. The expression of Cry1 increased by the intervention of Yupingfeng powder(P<0.05). |
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