Research On The Biorhythm Of Defensive Qi In TCM And The Primary Material Basis Involved

The theory of defensive qi, as an important part of TCM, has beenexpounded in Huangdi Neijing in its function and circulation. Much attentionhas been paid to this topic in TCM research, focusing on the origin, functionand clinical application. With the development of modern medical technique,some scholars tried to explain the theory of defensive qi in modern medicine.The theories about immune and thermometabolism are connected with thematerial basis of defensive qi. Though some aspects are not very certain, therelevant researches do contribute to the improvement of the theory ofdefensive qi. In a word, the modern researches of defensive qi explained thebasic material from its segmentary function, which going against the conceptof holism in TCM and original intention in Huangdi Neijing. Moreover, itrarely involved the research about the biorhythm of defensive qi.Metabonomics is a new branch of systematic biology after the rise ofgenomics, transcriptomics and proteomics. It is a kind of technology thatstudyies the metabolic network of biological systems by investigating themetabolites map and its dynamic change before or after the biological systemsbe stimulated or disturbed (such as a particular genetic mutation orenvironmental changes). Regardless of the complicated biochemicalmolecules interact with each other and changes in metabolic process,metabonomics has diversified and comprehensive analysis of the terminalproduct of metabolic as a whole and showed the changes of organic inner state,avoid using a single or a few indicators to research a certain pathological andphysiological changes. With the holistic, dynamic and comprehensivecharacteristic, this analysis method is in line with the integrity principle oftreating disease and herb effect in TCM.Using the platform of metabonomics, this paper explored the material basis of defensive qi and endogenous change rules from the daily, month andyear rhythm respectively, studied the effect of year rhythm of defensive qi oninterleukin10and interferonγin peripheral blood of healthy adults, anddiscussed the effect of "Jade screen Powder" on the rats with defensive qideficiency.This paper mainly includes the theory and experiment two parts:PART1THEORETICAL RESEARCHObjective: To explore biorhythm and material basis of defensive qi fromliterature.Method: According to the ancient and modern literature, discussed thesource, route, function,essence and new research way of defensive qi in detail.Conclusion: At present, researches of defensive qi only focus on a singlefunction to search the essential process of defensive qi, rather than explore thematerial method from the circadian rhythms of year, month, day and its quickand smooth characteristic, therefore it is difficult to interpret the essence ofdefensive qi, the range of material base of defensive qi should be researchfurther in an overall and comprehensive level, the technology platform ofmetabonomics provides the possibility.PART2EXPERIMENTAL RESEARCHEXPERIMENT1: Establishment of the model rats with defensive qideficiency and the scientific basis of metabonomics research of defensiveqiObjective: To find the potential biomarkers of deficiency syndrome ofdefensive qi by studying the characteristics of serum metabonomics in modelgroup and Chinese medicine group.Methods: Model group (n=10) of deficiency in defensive qi made byalternative hot and cold stimulation; the “jade screen decoction” be given tomake medication group (n=10); and a healthy control group (n=10). Systolicblood pressure, thymus index, WBC were measured in these groups. Serummetabolites were examined by Liquid chromatography-mass spectrometryinstrument, and PCA was used in analysis. Result: Systolic blood pressure of rats did not show obvious differenceamong these three groups (P>0.05); White blood cell count of model groupwas obviously higher than that of control group and medication group (P <0.05); thymus index of model group was obviously lower than that in controlgroup, medication group (P <0.05). Choline, phosphoryl Choline,L-Homoserine, L-Methionine, and Folic acid may be the potential biomarkersbetween control group and model group.Conclusion: Metabolic spectrum changes of rats of deficiency indefensive qi can be found by using the method of Serum metabonomics.Choline, phosphoryl choline, L-Homoserine, folic acid, L-methionine, couldbe the potential disease markers. Metabolic spectrum has actively changed inmodel rats with deficiency in defensive qi after intervened by jade screendecoction.EXPERIMENT2: Research on daily biorhythm of defensive qiObjective: To find the material basis of the daily biorhythm of defensiveqi, and explore its action mechanism from the angle of metabolic pathways,through the platform of metabonomicsMethod: Divide the SD rats (weights range from180g to200g) intoday group（12：00）and night group（24:00）according to random number tablemethod. At12:00/24:00respectively, measure their blood pressure, thymusweight and WBC. Serum metabolites were examined by Liquidchromatography-mass spectrometry instrument, and PCA was used inanalysis.Result: The blood pressure and WBC of rats were high in day and low innight (P <0.05), white thymus index was not statistically significant.4-methyl, dopamine, Phosphoryl choline and methionine do obviouscontribution to classification of day and night groups.Conclusion: Defensive qi has obvious daily rhythm,4-methyl catechol;Dopamine; Phosphoryl choline, methionine could be the potential biomarkersto influence the day rhythm changes of defensive qi.EXPERIMENT3: Research on month biorhythm of defensive qi Objective: To find the material basis of the month biorhythm ofdefensive qi, and explore its action mechanism from the angle of metabolicpathways, through the platform of metabonomicsMethod: Determine the early group (Chinese calendar on February22,2012), medium group (Chinese calendar March7,2012), and the late group(Chinese calendar March20,2012) according to the lunar calendar monthwhich has the spring equinox day. Divide the SD rats (weights range from180g to200g) into three groups (n=10) by the weight ascending orderaccording to the random number table method. At12:00on February22,2012,March7,2012and March20,2012, respectively, measure their bloodpressure, body weight, thymus weight and WBC. Serum metabolites wereexamined by Liquid chromatography-mass spectrometry instrument, andPCA was used in analysis.Result: The blood pressure and white blood cell count has rhythmchanges which is higher in middle group than in early group and late group (P<0.05). Thymus index of month rhythm is lower in the end of the month thanthan of other groups(P <0.05). Biomarkers which contribute to classificationof three groups are: Phosphoryl choline, L-Methionine, Serotonin, L-lysine.Conclusion: Defensive qi has obvious month rhythmic. Phosphorylcholine, L-Methionine, Serotonin, L-lysine may be the potential biomarkerswhich affect the monthly rhythmic change of defensive qi.EXPERIMENT4: Research on year biorhythm of defensive qiObjective: To find the material basis of the year biorhythm of defensiveqi, and explore its action mechanism from the angle of metabolic pathways,through the platform of metabonomics.Method: At about12:00on March30,2012(spring equinox), June21,2012(summer solstice) and September22,2012(autumn equinox),December21,2012(winter solstice), respectively, measure their blood pressure, bodyweight, thymus weight and WBC. Serum metabolites were examined byLiquid chromatography-mass spectrometry instrument, and PCA was usedin analysis. Result: The blood pressure, white blood cell count, thymus index of ratsshowed obvious seasonal fluctuations. In the winter solstice group, the rat’sblood pressure was significantly higher than the ones in summer solstice group(P <0.05); White blood cell count of rats in winter solstice group significantlylower than the ones in summer solstice group (P <0.05); Thymus index insummer solstice group is significantly higher than the ones in winter solsticegroup (P <0.05).Conclusion: Defensive qi has obvious year rhythm. Purine, erythrose,hyacinthin, methionine, Methoxytryptophan, threonine and homoserine areprobably the potential biomarkers which influence the year rhythmic changeof defensive qi.EXPERIMENT5: Impact of Year rhythm changes of defensive qi oninterleukin10and interferon γ in peripheral blood of healthy people.Objective: To observe the seasonal variation of immune markersinterleukin10and interferon γin the peripheral blood of healthy people.Method: Using enzyme-linked immunosorbent method to test the levelof interleukin10and interferon γ in the peripheral blood of20volunteers(10male and10female) respectively from the spring equinox group, thesummer solstice group, autumnal equinox group and winter solstice group.Result: The level of IFN-γin serum is4.58±0.21pg/L during thesummer solstice, which is significantly higher than that during the autumnalequinox（2.72±0.92）pg/L and winter solstic（e2.58±0.27pg/L）(P <0.05).The IL-10level during winter solstice is7.22+0.27pg/ml, which issignificantly higher than that during the summer solstice(5.89+0.33) pg/ml.(P <0.05). Index levels of male and female group in the four seasons have thesame trend of change respectively, there is no difference between the twogroups.Conclusion: The human immune markers IFN-γand interleukin10have obvious year rhythmic fluctuations.