卫气生理节律及相应物质基础变化规律的研究
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摘要
卫气理论是中医基础理论的重要组成部分。《黄帝内经》对卫气的功能及运行特点已有较为详尽的论述。卫气学说一直是中医的重要研究课题,一般集中在卫气的来源、运行、功能及其临床应用。近年来，随着现代医学技术的进步，一些学者尝试借助现代医学手段对卫气学说进行阐释。由此提出了关于卫气实质的一系列学说。主要包括：卫气的免疫学说，卫气的热代谢学说等，虽然在很多方面未取得一致的结论,却从某种程度上促进了卫气学说这一理论体系的完善。但关于卫气的现代研究大多从单一功能探求其相应物质基础，偏离了中医的整体观念和《黄帝内经》卫气理论的原意。而且，很少涉及到卫气的节律性研究。
代谢组学(metabolomics/metabonomics)是继基因组学、转录组学和蛋白质组学之后兴起的系统生物学的一个新的分支,它是通过考察生物体系受刺激或扰动前后(如将某个特定的基因变异或环境变化后)代谢产物图谱及其动态变化研究生物体系的代谢网络的一种技术。代谢组学抛开体内纷繁复杂的生化分子互相作用和网络式代谢过程变化,对代谢的终端产物进行多元化综合分析,从整体上展示生物体内在的变化状态,避免了以往采用单一指标或少数几个指标研究某种病理和生理变化。这种在方法学上具有融整体、动态、综合、分析于一体的特点,符合中医治疗疾病和中药作用的整体性原则。
本文借助代谢组学技术，依据《黄帝内经》对卫气的功能及节律性的描述，分别从卫气的日节律、月节律、年节律来探求卫气的物质基础和内源性变化规律，并讨论了卫气年节律对健康人群外周血白细胞介素10和干扰素γ的影响以及玉屏风散水煎剂对卫气虚大鼠的影响。
本篇论文内容主要包括理论和实验两大部分：
第一部分卫气的理论探讨
目的：从文献中发掘卫气的节律性变化规律及物质基础。
方法：根据古今文献记载，从卫气的来源、循行路线、功能、实质以及新研究方法的角度，对卫气进行了详细论述。
结论：目前卫气的现代研究仅着眼于单一功能探求卫气本质的过程，而忽略了从卫气的年、月、日的生理节律及慓疾滑利的运行特点探究其物质性的方法，因而难以诠释卫气的实质，对卫气的物质基础范围还缺乏整体而全面的认识和研究，代谢组学技术提供了这方面研究的可能性。
第二部分实验研究
实验一：卫气虚模型大鼠的建立及卫气相关物质基础的研究
目的：运用代谢组学技术研究卫气虚证大鼠及中药干预后的血清代谢组学特征，寻找卫气相关生物标志物。
方法：采用热寒交替刺激法制作卫气虚证大鼠模型（10只），给予玉屏风散水煎剂进行干预（10只），并设健康对照组（10只）。对三组大鼠，测量血压、计算胸腺指数，断头取血，测血常规；用液相色谱-质谱联用仪，对血清进行代谢物检测。利用主成分分析方法对数据进行统计分析。
结果：Table1-1, Fig.1-1示：大鼠血压在三组间未呈现明显差别（P＞0.05）；模型组的白细胞数明显高于对照组和治疗组（P<0.05）、模型组的胸腺指数明显低于对照组、治疗组（P＜0.05）。胆碱、磷酰胆碱、高丝氨酸、蛋氨酸、脱氧胞苷、叶酸可能是区分对照组与模型组差异的潜在生物标志物。
结论：运用血清代谢组学方法可以发现卫气虚证大鼠存在代谢谱的改变，胆碱、磷酰胆碱、高丝氨酸、蛋氨酸、叶酸可能是其潜在的疾病标志物。对卫气虚证大鼠应用玉屏风散水煎剂进行干预，其代谢谱具有积极的变化。
实验二：卫气日节律变化规律研究
目的：通过代谢组学技术，寻找卫气日节律变化规律的物质基础。
方法：按照随机数字表法，将体重约180-200g的SD大鼠随机分为日组（12：00）、夜组（24:00）每组10只。分别于12:00/24:00，测量血压、计算胸腺指数，断头取血，测血常规，用液相色谱-质谱联用仪，对血清进行代谢物检测。利用主成分分析方法对数据进行统计分析。
结果：大鼠血压、白细胞数均呈现明显的夜高昼低变化（P<0.05），胸腺指数的变化没有统计学意义（P＞0.05）。4-甲基邻苯二酚；多巴胺；磷酰胆碱、蛋氨酸对日夜两组分类有明显贡献。
结论：SD大鼠卫气存在昼夜节律变化，多巴胺、磷酰胆碱、蛋氨酸、4-甲基邻苯二酚可能是参与或调节卫气昼夜节律变化的潜在生物标志物。
实验三：卫气月节律变化规律研究
目的：通过代谢组学技术，寻找卫气月节律变化规律的物质基础。
方法：以春分日所在农历月，确定月初组（阳历2012年2月22日）、月中组（阳历2012年3月7日）、月末组（阳历2012年3月20日）。按照随机数字表法，将体重约180-200g的SD大鼠，称体重按升序排列，随机数字表法随机分入三组中（每组10只）。分别于2012年2月22日/2012年3月7日/2012年3月20日的12:00，测量血压，计算胸腺指数，断头取血，测血常规，用液相色谱-质谱联用仪，对血清进行代谢物检测。利用主成分分析方法对数据进行统计分析。
结果：大鼠血压、白细胞数、胸腺指数均呈现明显的月节律变化（P＜0.05）。大鼠血压、白细胞数月中组明显高于月初组、月末组（P＜0.05），月初组、月末组组间差异不明显（P＞0.05）。胸腺指数在月末组明显低于月初组、月中组。确定对三组分类有贡献的差异离子为：磷酰胆碱、蛋氨酸、血清素、赖氨酸。
结论：SD大鼠卫气存在月节律变化，磷酰胆碱、蛋氨酸、血清素、赖氨酸可能是参与或调节卫气月节律变化的潜在生物标志物。卫气的月节律变化可能影响到了机体氨基酸代谢。
实验四：卫气年节律变化规律研究
目的：通过代谢组学技术，寻找卫气年节律变化规律的物质基础。
方法：分别于春分日（2012年3月30日）、夏至日（2012年6月21日）、秋分日（2012年9月22日）、冬至日（2012年12月21日）的12:00左右，对春分、夏至、秋分、冬至四组大鼠（每组10只），测量血压，计算胸腺指数，断头取血，测血常规，用液相色谱-质谱联用仪，对血清进行代谢物检测。利用主成分分析方法对数据进行统计分析。
结果：大鼠血压、白细胞数、胸腺指数均表现出明显的季节性波动。大鼠血压在冬至组明显高于夏至组（P<0.05）；白细胞数在冬至组最低，与春分、夏至组均有统计学差异（P<0.05）、胸腺指数在夏至组明显高于其它组（P<0.05）。确定对春分、夏至、秋分、冬至四组间分类有贡献的差异离子有：蛋氨酸、磷酰胆碱、谷氨酰胺、低聚异麦芽糖、海藻糖。
结论：SD大鼠卫气存在年节律变化，磷酰胆碱、蛋氨酸、谷氨酰胺、低聚麦芽糖、海藻糖可能是参与或调节卫气年节律变化的潜在生物标志物。卫气的季节性变化可能对机体糖代谢产生了影响。
实验五：卫气的年节律对健康人外周血白介素10和干扰素γ的影响
目的：观察健康人群外周血清中人体免疫标志物白细胞介素10和干扰素γ的年节律性变化规律。
方法：分别在春分、夏至、秋分、冬至四个节气，采用酶联免疫吸附法检测春分组、夏至组、秋分组、冬至组健康志愿者（每组20名，男女各10名）外周血清中干扰素γ和白细胞介素10水平。
结果：血清中IFN-γ水平在夏至时（4.58±0.21） pg/L，明显高于秋分时（2.72±0.92）pg/L、冬至时（2.58±0.27） pg/L。（P<0.05）。 IL-10水平在冬至时（7.22±0.27）pg/ml，明显高于夏至时（5.89±0.33）pg/ml。（P<0.05）。男生组和女生组指标水平在四季分别有同样趋势变化，男女组之间没有差别。
结论：人体免疫标志物干扰素γ和白细胞介素10存在明显的年节律波动。
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 RESEARCH
Objective: 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 RESEARCH
EXPERIMENT1: Establishment of the model rats with defensive qideficiency and the scientific basis of metabonomics research of defensiveqi
Objective: 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 qi
Objective: 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 metabonomics
Method: 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 metabonomics
Method: 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 qi
Objective: 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.
代谢组学(metabolomics/metabonomics)是继基因组学、转录组学和蛋白质组学之后兴起的系统生物学的一个新的分支,它是通过考察生物体系受刺激或扰动前后(如将某个特定的基因变异或环境变化后)代谢产物图谱及其动态变化研究生物体系的代谢网络的一种技术。代谢组学抛开体内纷繁复杂的生化分子互相作用和网络式代谢过程变化,对代谢的终端产物进行多元化综合分析,从整体上展示生物体内在的变化状态,避免了以往采用单一指标或少数几个指标研究某种病理和生理变化。这种在方法学上具有融整体、动态、综合、分析于一体的特点,符合中医治疗疾病和中药作用的整体性原则。
本文借助代谢组学技术，依据《黄帝内经》对卫气的功能及节律性的描述，分别从卫气的日节律、月节律、年节律来探求卫气的物质基础和内源性变化规律，并讨论了卫气年节律对健康人群外周血白细胞介素10和干扰素γ的影响以及玉屏风散水煎剂对卫气虚大鼠的影响。
本篇论文内容主要包括理论和实验两大部分：
第一部分卫气的理论探讨
目的：从文献中发掘卫气的节律性变化规律及物质基础。
方法：根据古今文献记载，从卫气的来源、循行路线、功能、实质以及新研究方法的角度，对卫气进行了详细论述。
结论：目前卫气的现代研究仅着眼于单一功能探求卫气本质的过程，而忽略了从卫气的年、月、日的生理节律及慓疾滑利的运行特点探究其物质性的方法，因而难以诠释卫气的实质，对卫气的物质基础范围还缺乏整体而全面的认识和研究，代谢组学技术提供了这方面研究的可能性。
第二部分实验研究
实验一：卫气虚模型大鼠的建立及卫气相关物质基础的研究
目的：运用代谢组学技术研究卫气虚证大鼠及中药干预后的血清代谢组学特征，寻找卫气相关生物标志物。
方法：采用热寒交替刺激法制作卫气虚证大鼠模型（10只），给予玉屏风散水煎剂进行干预（10只），并设健康对照组（10只）。对三组大鼠，测量血压、计算胸腺指数，断头取血，测血常规；用液相色谱-质谱联用仪，对血清进行代谢物检测。利用主成分分析方法对数据进行统计分析。
结果：Table1-1, Fig.1-1示：大鼠血压在三组间未呈现明显差别（P＞0.05）；模型组的白细胞数明显高于对照组和治疗组（P<0.05）、模型组的胸腺指数明显低于对照组、治疗组（P＜0.05）。胆碱、磷酰胆碱、高丝氨酸、蛋氨酸、脱氧胞苷、叶酸可能是区分对照组与模型组差异的潜在生物标志物。
结论：运用血清代谢组学方法可以发现卫气虚证大鼠存在代谢谱的改变，胆碱、磷酰胆碱、高丝氨酸、蛋氨酸、叶酸可能是其潜在的疾病标志物。对卫气虚证大鼠应用玉屏风散水煎剂进行干预，其代谢谱具有积极的变化。
实验二：卫气日节律变化规律研究
目的：通过代谢组学技术，寻找卫气日节律变化规律的物质基础。
方法：按照随机数字表法，将体重约180-200g的SD大鼠随机分为日组（12：00）、夜组（24:00）每组10只。分别于12:00/24:00，测量血压、计算胸腺指数，断头取血，测血常规，用液相色谱-质谱联用仪，对血清进行代谢物检测。利用主成分分析方法对数据进行统计分析。
结果：大鼠血压、白细胞数均呈现明显的夜高昼低变化（P<0.05），胸腺指数的变化没有统计学意义（P＞0.05）。4-甲基邻苯二酚；多巴胺；磷酰胆碱、蛋氨酸对日夜两组分类有明显贡献。
结论：SD大鼠卫气存在昼夜节律变化，多巴胺、磷酰胆碱、蛋氨酸、4-甲基邻苯二酚可能是参与或调节卫气昼夜节律变化的潜在生物标志物。
实验三：卫气月节律变化规律研究
目的：通过代谢组学技术，寻找卫气月节律变化规律的物质基础。
方法：以春分日所在农历月，确定月初组（阳历2012年2月22日）、月中组（阳历2012年3月7日）、月末组（阳历2012年3月20日）。按照随机数字表法，将体重约180-200g的SD大鼠，称体重按升序排列，随机数字表法随机分入三组中（每组10只）。分别于2012年2月22日/2012年3月7日/2012年3月20日的12:00，测量血压，计算胸腺指数，断头取血，测血常规，用液相色谱-质谱联用仪，对血清进行代谢物检测。利用主成分分析方法对数据进行统计分析。
结果：大鼠血压、白细胞数、胸腺指数均呈现明显的月节律变化（P＜0.05）。大鼠血压、白细胞数月中组明显高于月初组、月末组（P＜0.05），月初组、月末组组间差异不明显（P＞0.05）。胸腺指数在月末组明显低于月初组、月中组。确定对三组分类有贡献的差异离子为：磷酰胆碱、蛋氨酸、血清素、赖氨酸。
结论：SD大鼠卫气存在月节律变化，磷酰胆碱、蛋氨酸、血清素、赖氨酸可能是参与或调节卫气月节律变化的潜在生物标志物。卫气的月节律变化可能影响到了机体氨基酸代谢。
实验四：卫气年节律变化规律研究
目的：通过代谢组学技术，寻找卫气年节律变化规律的物质基础。
方法：分别于春分日（2012年3月30日）、夏至日（2012年6月21日）、秋分日（2012年9月22日）、冬至日（2012年12月21日）的12:00左右，对春分、夏至、秋分、冬至四组大鼠（每组10只），测量血压，计算胸腺指数，断头取血，测血常规，用液相色谱-质谱联用仪，对血清进行代谢物检测。利用主成分分析方法对数据进行统计分析。
结果：大鼠血压、白细胞数、胸腺指数均表现出明显的季节性波动。大鼠血压在冬至组明显高于夏至组（P<0.05）；白细胞数在冬至组最低，与春分、夏至组均有统计学差异（P<0.05）、胸腺指数在夏至组明显高于其它组（P<0.05）。确定对春分、夏至、秋分、冬至四组间分类有贡献的差异离子有：蛋氨酸、磷酰胆碱、谷氨酰胺、低聚异麦芽糖、海藻糖。
结论：SD大鼠卫气存在年节律变化，磷酰胆碱、蛋氨酸、谷氨酰胺、低聚麦芽糖、海藻糖可能是参与或调节卫气年节律变化的潜在生物标志物。卫气的季节性变化可能对机体糖代谢产生了影响。
实验五：卫气的年节律对健康人外周血白介素10和干扰素γ的影响
目的：观察健康人群外周血清中人体免疫标志物白细胞介素10和干扰素γ的年节律性变化规律。
方法：分别在春分、夏至、秋分、冬至四个节气，采用酶联免疫吸附法检测春分组、夏至组、秋分组、冬至组健康志愿者（每组20名，男女各10名）外周血清中干扰素γ和白细胞介素10水平。
结果：血清中IFN-γ水平在夏至时（4.58±0.21） pg/L，明显高于秋分时（2.72±0.92）pg/L、冬至时（2.58±0.27） pg/L。（P<0.05）。 IL-10水平在冬至时（7.22±0.27）pg/ml，明显高于夏至时（5.89±0.33）pg/ml。（P<0.05）。男生组和女生组指标水平在四季分别有同样趋势变化，男女组之间没有差别。
结论：人体免疫标志物干扰素γ和白细胞介素10存在明显的年节律波动。
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 RESEARCH
Objective: 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 RESEARCH
EXPERIMENT1: Establishment of the model rats with defensive qideficiency and the scientific basis of metabonomics research of defensiveqi
Objective: 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 qi
Objective: 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 metabonomics
Method: 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 metabonomics
Method: 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 qi
Objective: 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.
引文
1姚春鹏译注,黄帝内经.北京:中华医书局,2010
2章恪.卫气——免疫细胞的中医表达形式.湖北中医杂志,2001,23(3):3
3周晓平,杨进.从气象医学理论探析卫气的实质.中国中医基础医学杂志.2007,13(4):259-260
4杨上善撰,王洪图,李云校.黄帝内经太素(第三版).北京:科技文献出版社,2013
5扁鹊撰,柴铁劬校注.难经.北京:科技文献出版社,2010
6皇甫谧撰,周琦校注.针灸甲乙经.北京:中国医药科技出版社,2011
7张志聪撰,孙国中,方向红校注.黄帝内经素问集注(修订本)学苑出版社.2011
8孙思邈撰.备急千金要方.太原:山西科学技术出版社,2010
9华佗撰,农汉才校注.中藏经.北京:学苑出版社,2007
10张丽娜,黄晶,杨旭."卫出于上焦"辨识.中医药学报.2004,32(6):35-37
11匡耀祖.略论卫气出于下焦.江西中医药,1985,(1):3-5
12靳士华.试论《灵枢》"营卫"经义.天津中医,1991,(2):38.
13张景岳撰,景岳全书.太原:山西科学技术出版社,2006
14汪幼李.论"卫出下焦".中医杂志,1981,(12):12
15赵昆霞.论"卫出下焦".云南中医学院学报.2005,28(2):24-25
16郑宇东.论卫出于肾.河南中医药学刊.1999,14(4):3-5
17章恪.卫气的来源与分布探析.中医药学刊.2002,20(2):205,234
18李霜.从"三焦出气"浅谈营气、卫气.新疆中医药.2005,23(5):1
19吴弥漫.《内经》卫气学说的整理及研究.北京中医学院学报,1992,15(2):12
20吴萍.谈《内经》卫气运行.四川中医,2004,22(2):20-21.
21白凌志.卫气运行与人体节律.黑龙江中医药,1987,(5):4-5
22区永欣.卫气生理病理的研究.中医药学报,1989,5(5):20
23卢志刚,王亚利,张明泉,等.季节变化对健康人免疫功能的影响.第四军医大学学报,2009,30(13):1209-1211
24张明泉,王亚利,郭霞珍,等.褪黑素在中医学天人相应理论中的地位和作用.新中医,2009,41(3):10-11
25卢志刚,王亚利,张明泉,等.冬夏季节变化对健康人外周血免疫淋巴细胞的影响.北京中医药大学学报,2010,33(2):106-109
26卢志刚,王亚利,张明泉,等.秋冬季节对健康大鼠肺组织表面活性蛋白A和白细胞介素6表达的影响.中国组织工程研究与临床康复,2010,14(11):2000-2003
27师旭亮,王亚利,张明泉,等.从“天人相应”探讨季节变化对健康人外周血干扰素γ和白细胞介素10的影响.中医杂志,2013,54(7):573-575
28秦树仙.营卫实质再探.云南中医学院学报,1993,16(3):7-8
29王洪琦,李建国,马晶,等.卫气虚弱与机体免疫机制相关性研究.中国中医药信息杂志,2003,10(12):12
30许朝进,席孝贤,贺新怀.卫气与黏膜免疫的相关性探讨.中医药学刊,2004,22(12):12
31张越,陈扬荣.卫气营血的免疫相关性探讨.中医研究,2007,20(12):l-2
32李忠正,郭义,郭永明.浅析卫气与血管外体液循环系统的关系.中医药学报.2010,38(4):1-2
33陈萌.卫气的实质与医学意义.中华中医药杂志,2007,22(8):8
34Gennan JB,Bauman DE,Burrrin D6,et a1.Metabolomics in the openingdecade of the21st century: building the roads to individualizedhealth.Nutn,2004,134(10):2729
35Nicholson J K,Holmes E,Lindon J C,et a1.The challenges of modelingmammalian biocomplexity.Nat Biotechnol,2004,22(10):1268-1274
1郑筱萸主编.中药新药临床研究指导原则.北京:中国医药科技出版社.2002:390
2徐婧,田亚平,陈艳华,等,代谢组学血浆样品前处理及其快速高分辨液相色谱-质谱分析方法研究.分析化学研究报告,2011,39(12):1793-1797
3方素萍.卫益颗粒(玉屏风散)对小鼠冷热刺激适应能力的影响.中国实验方剂学杂志,2003,9(3):33-35
4陈新.加味玉屏风汤对截肢应激小鼠细胞免疫功能的调节作用.中医杂志,1998,39(12):746
5李伯南.加味玉屏风散对手术应激小鼠胸腺超微结构的影响.中国病理生理杂志,1996,12(2):178-180
6刘福岭.主编.现代医学辞典.济南:山东科学技术出版社.1990:80
7杨永明,卢德勋,甄玉国.胆碱及其作用.饲料博览,2001,(3):38-40
8刘福岭主编.现代医学辞典.济南:山东科学技术出版社.1990:84
9卢广林,李东风.蛋氨酸螯合锌对野鸭免疫器官及血液指标的影响.饲料研究,2011,(8):40-41
10张明泉,王亚利,郭霞珍,等.褪黑素在中医学天人相应理论中的地位和作用.新中医,2009,41(3)10-11
11刘福岭.主编.现代医学辞典.济南:山东科学技术出版社.1990:695
12钟少华,何坚,郭晓玲,等.大豆异黄酮和叶酸对同型半胱氨酸糖尿病动物模型免疫功能的影响.新乡医学院学报,2008,25(2):190-192
1GuoYE, Stein PK. Circadian Rhythm in the Cardiovascular System:Chronoeardiology.Am Heart J,2003,145:779-786
2Degaute JP, van de Borne P, Linkowski P,et al.Quantitative analysis ofthe24-hour blood pressure and heart rate patterns in young men.Hypertension,1991,18:199-210
3Hu K, Ivanov PC,Hilton MF, et al.Endogenous circadian rhythm in anindex of cardiac vulnerability independent of changes in behavior. ProcNatl Acad Sci USA,2004,101(4):18223-18227
4James A.Shaw, Jaye P.F. Chin-Dusting, Bronwyn A,et al.DiurnalVariation in Endothelium-Dependent Vasodilatation Is Not Apparent inCornary Artery Disease.Circulation,2001,103(6):806-812
5Menon IS,Smith PA,White RW.Diurnal variations of fibrinolytic activityand plasma-11-hydroxycorticosteroid levels.Lancet,1967,2:531-533
6Faiman C, Moorhouse JA.Diurnal variation in the level of glucose andrelated substances in healthy and diabetic subjects during starvation.Clin.Sci,1967,32:111-126
7吴萍.谈《内经》营卫之气与睡眠.时珍国医国药,2010,21(7):1840-1841
8黄俊山,樊建锋,刘芳,等.失眠患者血清甲状腺激素及多巴胺水平与中医证型的关系.中国中西医结合杂志,2011,31(3):338-341
9徐婧,田亚平,陈艳华,等,代谢组学血浆样品前处理及其快速高分辨液相色谱-质谱分析方法研究.分析化学研究报告,2011,39(12):1793-1797
10童建.免疫系统的生物节律.国外医学免疫学分册,1999,22(1):30-33
1马孔琛,李经才,李彩霞.胰岛素降小鼠血糖作用的月节律.沈阳药科大学学报,1996,13(4):298-299
2丛鹏,陈璐,房莉萍.心率变异性存在月节律现象.北京中医药,2011,30(7):508-509
3三宅良明.28天月经周期与妊娠时的生物节律.日本医学介绍,2000,21(2):65-67
4冷闻辉,于明,朱颖,等.盐酸氟桂利嗪胶囊治疗月经期偏头痛的临床疗效观察.脑与神经疾病杂志,2011,19(2):107-109
5夏桂成.月周期节律诱导法调理月经周期.南京中医药大学学报（自然科学版）,2000,16(1):11-12
6徐婧,田亚平,陈艳华,等.代谢组学血浆样品前处理及其快速高分辨液相色谱-质谱分析方法研究.分析化学研究报告,2011,39(12):1793-1797
7何裕民.月亮盈亏与人体机能.北京中医学院学报.1986,2(2):20
8王敏.金春玉,毛军,等.对《黄帝内经》月节律的科学性探索——通过对中药马钱子、黄连毒性的观察，验证《黄帝内经》月节律理论的科学性.中国中医基础医学杂志,2001,7(12):25-27
9夏质彬.运用人体月节律相月诊治疾病的临床探讨.中医研究.2007,20(8):12-14
10马星,刘莎,岳武林,等.月球月相对精神情绪行为影响探讨.医学理论与实践,1994,7(5):40-41
1卢志刚,王亚利,张明泉,等.秋冬季节对健康大鼠肺组织表面活性蛋白A和白细胞介素6表达的影响.中国组织工程研究与临床康复,2010,14(11):2000-2003
2卢一.谷胱甘肽的生物节律研究进展.国外医学卫生学分册,1999,26(5):261-262
3王洁琼,张蕴琨.谷氨酸、生物节律与运动.南京体育学院学报(自然科学版),2007,6(2):42-46
4张鹏,林才,徐建军.谷氨酰胺双肽对严重烧伤患者T淋巴细胞亚群的影响.中国临床营养杂志,2006,14(1):15-17
5卢志刚,王亚利,张明泉,等.冬夏季节变化对健康人外周血免疫淋巴细胞的影响.北京中医药大学学报,2010,33(2):106-109
6胡跃.四川食品发酵,1992,(1):26-28
1李祥华,涂献玉,张德新,等.五味地龙汤对哮喘豚鼠炎症细胞核细胞因子的影响.中医杂志.2008,49(6):543-545
2左慧敏,刘秀云,江载芳.白介素8白介素10及γ-干扰素在肺炎支原体肺炎中的作用.中国实用儿科杂志.2008,23(4):269-271
3刘建,周朕,刘晓晴.外周血单个核细胞端粒酶活性及IL-2、IL-12水平与老年性肺炎关系的研究.内科急危重症杂志,2006,12(5):221-222
4王颖超,徐学聚,郭宏湘,等.支气管哮喘患儿血浆白细胞介素-6γ-干扰素及体液免疫变化探讨.医药论坛杂志.2006,27(23):60-61
5王喜英,吴波,张愔.白细胞介素-10在慢性阻塞性肺病检测中的意义.医药论坛杂志,2006,27(24):19-20
6阎星诗,洪素兰.中医时间医学研究述评.世界中西医结合杂志.2009,4(2):143-144
7申维玺,孙燕.论中医证的化学本质是蛋白质和肽及证本质的分子标准.中国中西医结合杂志.1999,19(11):696-698
8申维玺.再论中医证的本质是细胞因子.中医杂志.2002,43(12):888-891
9Hoffmann R，Wenzel E，Huth A，et a1．Cytokine mRNA levels inalopecia areata before and after treatment with contact allergendiphenylcyclopropenone．J Investigative Dermatology，1994，103(4)：530-533
10Grunig G, Ford JG, Donaldson DD, et al. Roles of interleukin-13andinterferon-gamma in lung inflammation. J Chest,2002,121(3):88-95
11Fonseca-Aten M，Katz K，Hardy RD，et a1．Mycoplasma pneumoniaeinduces host-dependent pulmonary inflammation and airwayobstruction in mice．American J Respiratory Cell Molecular Biology,2005，32(3)：201-210
1Kitano H. Systems biology: a brief overview. Science,2002,295(5560):1662-1664
2Gates SC, Sweeley CC. Quantitative metabolic profiling based on gaschromatography. Clin Chem,1978,24(10)1663-1673
3Nicholson JK, Lindon JC, Holmes E.“Metabonomics”: understandingthe metabolic responses of living systems to pathophysiological stimulivia multivariate statistical analysis of biological NMR spectroscopicdata. Xenobiotica,1999,29(11)1181-1189
4Fiehn O, Kopka J, Dormann P, etal. Metabolite profiling for plantfunctional genomics. Nat Biotechnol,2000,18(11):1157-1161
5Field D, Sansone SA. A special issue on data standards. Omics,2006,10(2):84-93
6Griffin JL. Metabonomics: NMR spectroscopy and pattern recognitionanalysis of body fluids and tissues for characterization of xenobiotictoxicity and disease diagnosis. Curr Opin Chem Biol,2003,7(5)648-654
7Holmes E, Nicholson JK, Tranter G. Metabonomic characterization ofgenetic variations in toxicological and metabolic responses usingprobabilistic neural networks.Chem Res Toxicol,2001,14(2)546-551
8刘卫红，张琪，颜贤忠等，高脂血症及动脉粥样硬化痰瘀演变的代谢组学研究.中医杂志，2008，49（8）：738-741
9林云径，汪晋，王毅等，气相色谱-质谱法分析膀胱癌患者尿液代谢组.分析化学研究简报，2008，36（9）：1257-1260
10Glinski M, Weckwerth W. The role of mass spectrometry in plantsystems biology. Mass Spectrom Rev,2006,25(2):173-214
11Styczynski M P, Moxley JF, Tong LV, etal. Systematic identificationof conserved metabolites in GC/MS data for metabolomics andbiomarker discovery.Anal Chem,2007,79(3)966-973
12Yang J, Zhao XJ, Liu XL, etal. High performance liquidchromatography-mass spectrometry for metabonomics:potentialbiomarkers for acute deterioration of liver function in chronic hepatitisB[J]. J Proteome Res,2006,5(3):554-561
13SO GA T, OHASHIY, UENO Y, etal. Joural of Proteome Research,2003,2(5):488-494
14沈自尹.从肾本质研究到证本质研究的思考与实践上海中医药杂志.2000，3（4）：4-7
15Chen MJ, Zhao LP, Jia W, etal. Metabonomic study on the biochemicalprofiles of a hydrocortisone induced animal model..J Proteome Res,2005,4(6):2391-2396
16罗和古，丁杰，岳广欣，等.大鼠肝郁脾虚证的代谢组学研究.中西医结合学报，2007,5（3）：307
17Clayton TA, Lindon JC, Cloarec O, et a1Pharmaco－metabonomicphenotyping and personalized drug treatment. Nature,2006,440(7087):1073－1077
18Rasmussen B, Cloarec O,Tand HR, etal Multivariate analysis ofintegrated and full-resolution H-NMR spectral data from complexpharmaceutical preparations: St John's wort[J].PlantaMed,2006,72:556-563
19Nicholson JK, Connelly J,Lindon JC,etal.Metabonomics: a platform forstudying drug toxicity and gene function[J].Nat Rev DrugDiscov,2002,1(2)153-161
20John C, Lindon, Elaine Holmes, Jeremy K Nicholson. Toxicologicalapplications of magnetic resonance[J]Prog NMRspectrose,2004,45:109-143
2章恪.卫气——免疫细胞的中医表达形式.湖北中医杂志,2001,23(3):3
3周晓平,杨进.从气象医学理论探析卫气的实质.中国中医基础医学杂志.2007,13(4):259-260
4杨上善撰,王洪图,李云校.黄帝内经太素(第三版).北京:科技文献出版社,2013
5扁鹊撰,柴铁劬校注.难经.北京:科技文献出版社,2010
6皇甫谧撰,周琦校注.针灸甲乙经.北京:中国医药科技出版社,2011
7张志聪撰,孙国中,方向红校注.黄帝内经素问集注(修订本)学苑出版社.2011
8孙思邈撰.备急千金要方.太原:山西科学技术出版社,2010
9华佗撰,农汉才校注.中藏经.北京:学苑出版社,2007
10张丽娜,黄晶,杨旭."卫出于上焦"辨识.中医药学报.2004,32(6):35-37
11匡耀祖.略论卫气出于下焦.江西中医药,1985,(1):3-5
12靳士华.试论《灵枢》"营卫"经义.天津中医,1991,(2):38.
13张景岳撰,景岳全书.太原:山西科学技术出版社,2006
14汪幼李.论"卫出下焦".中医杂志,1981,(12):12
15赵昆霞.论"卫出下焦".云南中医学院学报.2005,28(2):24-25
16郑宇东.论卫出于肾.河南中医药学刊.1999,14(4):3-5
17章恪.卫气的来源与分布探析.中医药学刊.2002,20(2):205,234
18李霜.从"三焦出气"浅谈营气、卫气.新疆中医药.2005,23(5):1
19吴弥漫.《内经》卫气学说的整理及研究.北京中医学院学报,1992,15(2):12
20吴萍.谈《内经》卫气运行.四川中医,2004,22(2):20-21.
21白凌志.卫气运行与人体节律.黑龙江中医药,1987,(5):4-5
22区永欣.卫气生理病理的研究.中医药学报,1989,5(5):20
23卢志刚,王亚利,张明泉,等.季节变化对健康人免疫功能的影响.第四军医大学学报,2009,30(13):1209-1211
24张明泉,王亚利,郭霞珍,等.褪黑素在中医学天人相应理论中的地位和作用.新中医,2009,41(3):10-11
25卢志刚,王亚利,张明泉,等.冬夏季节变化对健康人外周血免疫淋巴细胞的影响.北京中医药大学学报,2010,33(2):106-109
26卢志刚,王亚利,张明泉,等.秋冬季节对健康大鼠肺组织表面活性蛋白A和白细胞介素6表达的影响.中国组织工程研究与临床康复,2010,14(11):2000-2003
27师旭亮,王亚利,张明泉,等.从“天人相应”探讨季节变化对健康人外周血干扰素γ和白细胞介素10的影响.中医杂志,2013,54(7):573-575
28秦树仙.营卫实质再探.云南中医学院学报,1993,16(3):7-8
29王洪琦,李建国,马晶,等.卫气虚弱与机体免疫机制相关性研究.中国中医药信息杂志,2003,10(12):12
30许朝进,席孝贤,贺新怀.卫气与黏膜免疫的相关性探讨.中医药学刊,2004,22(12):12
31张越,陈扬荣.卫气营血的免疫相关性探讨.中医研究,2007,20(12):l-2
32李忠正,郭义,郭永明.浅析卫气与血管外体液循环系统的关系.中医药学报.2010,38(4):1-2
33陈萌.卫气的实质与医学意义.中华中医药杂志,2007,22(8):8
34Gennan JB,Bauman DE,Burrrin D6,et a1.Metabolomics in the openingdecade of the21st century: building the roads to individualizedhealth.Nutn,2004,134(10):2729
35Nicholson J K,Holmes E,Lindon J C,et a1.The challenges of modelingmammalian biocomplexity.Nat Biotechnol,2004,22(10):1268-1274
1郑筱萸主编.中药新药临床研究指导原则.北京:中国医药科技出版社.2002:390
2徐婧,田亚平,陈艳华,等,代谢组学血浆样品前处理及其快速高分辨液相色谱-质谱分析方法研究.分析化学研究报告,2011,39(12):1793-1797
3方素萍.卫益颗粒(玉屏风散)对小鼠冷热刺激适应能力的影响.中国实验方剂学杂志,2003,9(3):33-35
4陈新.加味玉屏风汤对截肢应激小鼠细胞免疫功能的调节作用.中医杂志,1998,39(12):746
5李伯南.加味玉屏风散对手术应激小鼠胸腺超微结构的影响.中国病理生理杂志,1996,12(2):178-180
6刘福岭.主编.现代医学辞典.济南:山东科学技术出版社.1990:80
7杨永明,卢德勋,甄玉国.胆碱及其作用.饲料博览,2001,(3):38-40
8刘福岭主编.现代医学辞典.济南:山东科学技术出版社.1990:84
9卢广林,李东风.蛋氨酸螯合锌对野鸭免疫器官及血液指标的影响.饲料研究,2011,(8):40-41
10张明泉,王亚利,郭霞珍,等.褪黑素在中医学天人相应理论中的地位和作用.新中医,2009,41(3)10-11
11刘福岭.主编.现代医学辞典.济南:山东科学技术出版社.1990:695
12钟少华,何坚,郭晓玲,等.大豆异黄酮和叶酸对同型半胱氨酸糖尿病动物模型免疫功能的影响.新乡医学院学报,2008,25(2):190-192
1GuoYE, Stein PK. Circadian Rhythm in the Cardiovascular System:Chronoeardiology.Am Heart J,2003,145:779-786
2Degaute JP, van de Borne P, Linkowski P,et al.Quantitative analysis ofthe24-hour blood pressure and heart rate patterns in young men.Hypertension,1991,18:199-210
3Hu K, Ivanov PC,Hilton MF, et al.Endogenous circadian rhythm in anindex of cardiac vulnerability independent of changes in behavior. ProcNatl Acad Sci USA,2004,101(4):18223-18227
4James A.Shaw, Jaye P.F. Chin-Dusting, Bronwyn A,et al.DiurnalVariation in Endothelium-Dependent Vasodilatation Is Not Apparent inCornary Artery Disease.Circulation,2001,103(6):806-812
5Menon IS,Smith PA,White RW.Diurnal variations of fibrinolytic activityand plasma-11-hydroxycorticosteroid levels.Lancet,1967,2:531-533
6Faiman C, Moorhouse JA.Diurnal variation in the level of glucose andrelated substances in healthy and diabetic subjects during starvation.Clin.Sci,1967,32:111-126
7吴萍.谈《内经》营卫之气与睡眠.时珍国医国药,2010,21(7):1840-1841
8黄俊山,樊建锋,刘芳,等.失眠患者血清甲状腺激素及多巴胺水平与中医证型的关系.中国中西医结合杂志,2011,31(3):338-341
9徐婧,田亚平,陈艳华,等,代谢组学血浆样品前处理及其快速高分辨液相色谱-质谱分析方法研究.分析化学研究报告,2011,39(12):1793-1797
10童建.免疫系统的生物节律.国外医学免疫学分册,1999,22(1):30-33
1马孔琛,李经才,李彩霞.胰岛素降小鼠血糖作用的月节律.沈阳药科大学学报,1996,13(4):298-299
2丛鹏,陈璐,房莉萍.心率变异性存在月节律现象.北京中医药,2011,30(7):508-509
3三宅良明.28天月经周期与妊娠时的生物节律.日本医学介绍,2000,21(2):65-67
4冷闻辉,于明,朱颖,等.盐酸氟桂利嗪胶囊治疗月经期偏头痛的临床疗效观察.脑与神经疾病杂志,2011,19(2):107-109
5夏桂成.月周期节律诱导法调理月经周期.南京中医药大学学报（自然科学版）,2000,16(1):11-12
6徐婧,田亚平,陈艳华,等.代谢组学血浆样品前处理及其快速高分辨液相色谱-质谱分析方法研究.分析化学研究报告,2011,39(12):1793-1797
7何裕民.月亮盈亏与人体机能.北京中医学院学报.1986,2(2):20
8王敏.金春玉,毛军,等.对《黄帝内经》月节律的科学性探索——通过对中药马钱子、黄连毒性的观察，验证《黄帝内经》月节律理论的科学性.中国中医基础医学杂志,2001,7(12):25-27
9夏质彬.运用人体月节律相月诊治疾病的临床探讨.中医研究.2007,20(8):12-14
10马星,刘莎,岳武林,等.月球月相对精神情绪行为影响探讨.医学理论与实践,1994,7(5):40-41
1卢志刚,王亚利,张明泉,等.秋冬季节对健康大鼠肺组织表面活性蛋白A和白细胞介素6表达的影响.中国组织工程研究与临床康复,2010,14(11):2000-2003
2卢一.谷胱甘肽的生物节律研究进展.国外医学卫生学分册,1999,26(5):261-262
3王洁琼,张蕴琨.谷氨酸、生物节律与运动.南京体育学院学报(自然科学版),2007,6(2):42-46
4张鹏,林才,徐建军.谷氨酰胺双肽对严重烧伤患者T淋巴细胞亚群的影响.中国临床营养杂志,2006,14(1):15-17
5卢志刚,王亚利,张明泉,等.冬夏季节变化对健康人外周血免疫淋巴细胞的影响.北京中医药大学学报,2010,33(2):106-109
6胡跃.四川食品发酵,1992,(1):26-28
1李祥华,涂献玉,张德新,等.五味地龙汤对哮喘豚鼠炎症细胞核细胞因子的影响.中医杂志.2008,49(6):543-545
2左慧敏,刘秀云,江载芳.白介素8白介素10及γ-干扰素在肺炎支原体肺炎中的作用.中国实用儿科杂志.2008,23(4):269-271
3刘建,周朕,刘晓晴.外周血单个核细胞端粒酶活性及IL-2、IL-12水平与老年性肺炎关系的研究.内科急危重症杂志,2006,12(5):221-222
4王颖超,徐学聚,郭宏湘,等.支气管哮喘患儿血浆白细胞介素-6γ-干扰素及体液免疫变化探讨.医药论坛杂志.2006,27(23):60-61
5王喜英,吴波,张愔.白细胞介素-10在慢性阻塞性肺病检测中的意义.医药论坛杂志,2006,27(24):19-20
6阎星诗,洪素兰.中医时间医学研究述评.世界中西医结合杂志.2009,4(2):143-144
7申维玺,孙燕.论中医证的化学本质是蛋白质和肽及证本质的分子标准.中国中西医结合杂志.1999,19(11):696-698
8申维玺.再论中医证的本质是细胞因子.中医杂志.2002,43(12):888-891
9Hoffmann R，Wenzel E，Huth A，et a1．Cytokine mRNA levels inalopecia areata before and after treatment with contact allergendiphenylcyclopropenone．J Investigative Dermatology，1994，103(4)：530-533
10Grunig G, Ford JG, Donaldson DD, et al. Roles of interleukin-13andinterferon-gamma in lung inflammation. J Chest,2002,121(3):88-95
11Fonseca-Aten M，Katz K，Hardy RD，et a1．Mycoplasma pneumoniaeinduces host-dependent pulmonary inflammation and airwayobstruction in mice．American J Respiratory Cell Molecular Biology,2005，32(3)：201-210
1Kitano H. Systems biology: a brief overview. Science,2002,295(5560):1662-1664
2Gates SC, Sweeley CC. Quantitative metabolic profiling based on gaschromatography. Clin Chem,1978,24(10)1663-1673
3Nicholson JK, Lindon JC, Holmes E.“Metabonomics”: understandingthe metabolic responses of living systems to pathophysiological stimulivia multivariate statistical analysis of biological NMR spectroscopicdata. Xenobiotica,1999,29(11)1181-1189
4Fiehn O, Kopka J, Dormann P, etal. Metabolite profiling for plantfunctional genomics. Nat Biotechnol,2000,18(11):1157-1161
5Field D, Sansone SA. A special issue on data standards. Omics,2006,10(2):84-93
6Griffin JL. Metabonomics: NMR spectroscopy and pattern recognitionanalysis of body fluids and tissues for characterization of xenobiotictoxicity and disease diagnosis. Curr Opin Chem Biol,2003,7(5)648-654
7Holmes E, Nicholson JK, Tranter G. Metabonomic characterization ofgenetic variations in toxicological and metabolic responses usingprobabilistic neural networks.Chem Res Toxicol,2001,14(2)546-551
8刘卫红，张琪，颜贤忠等，高脂血症及动脉粥样硬化痰瘀演变的代谢组学研究.中医杂志，2008，49（8）：738-741
9林云径，汪晋，王毅等，气相色谱-质谱法分析膀胱癌患者尿液代谢组.分析化学研究简报，2008，36（9）：1257-1260
10Glinski M, Weckwerth W. The role of mass spectrometry in plantsystems biology. Mass Spectrom Rev,2006,25(2):173-214
11Styczynski M P, Moxley JF, Tong LV, etal. Systematic identificationof conserved metabolites in GC/MS data for metabolomics andbiomarker discovery.Anal Chem,2007,79(3)966-973
12Yang J, Zhao XJ, Liu XL, etal. High performance liquidchromatography-mass spectrometry for metabonomics:potentialbiomarkers for acute deterioration of liver function in chronic hepatitisB[J]. J Proteome Res,2006,5(3):554-561
13SO GA T, OHASHIY, UENO Y, etal. Joural of Proteome Research,2003,2(5):488-494
14沈自尹.从肾本质研究到证本质研究的思考与实践上海中医药杂志.2000，3（4）：4-7
15Chen MJ, Zhao LP, Jia W, etal. Metabonomic study on the biochemicalprofiles of a hydrocortisone induced animal model..J Proteome Res,2005,4(6):2391-2396
16罗和古，丁杰，岳广欣，等.大鼠肝郁脾虚证的代谢组学研究.中西医结合学报，2007,5（3）：307
17Clayton TA, Lindon JC, Cloarec O, et a1Pharmaco－metabonomicphenotyping and personalized drug treatment. Nature,2006,440(7087):1073－1077
18Rasmussen B, Cloarec O,Tand HR, etal Multivariate analysis ofintegrated and full-resolution H-NMR spectral data from complexpharmaceutical preparations: St John's wort[J].PlantaMed,2006,72:556-563
19Nicholson JK, Connelly J,Lindon JC,etal.Metabonomics: a platform forstudying drug toxicity and gene function[J].Nat Rev DrugDiscov,2002,1(2)153-161
20John C, Lindon, Elaine Holmes, Jeremy K Nicholson. Toxicologicalapplications of magnetic resonance[J]Prog NMRspectrose,2004,45:109-143
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