补中益气丸干预气虚质的临床观察及其血浆代谢组学机制研究
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摘要
背景：中医体质是指人体生命过程中,在先天禀赋和后天获得的基础上所形成的形态结构、生理功能和心理状态方面综合的、相对稳定的固有特质；表现为结构、功能、代谢以及对外界刺激反应等方面的个体差异性,对某些病因和疾病的易感性,以及疾病传变转归中的某种倾向性。而气虚质则是以气短懒言,容易疲乏,精神不振,易出汗等为主要表现的体质类型。
目前对气虚质的研究仍处于流行病学调查阶段,其基因、蛋白、代谢方面的机制研究并不多。根据本课题组前期对体质的研究结果发现：①广东地区健康人群以平和质居多(占81.3%),偏颇体质仅占18.7%；亚健康人群以偏颇体质居多(占64.0%),其中气虚质所占比例最大(占41%)。②对健康、亚健康人群的中医体质进行Logistic回归发现,平和质与健康呈正相关,与亚健康呈负相关,气虚质与亚健康呈正相关。③平和质的人,不容易出现亚健康、疾病；偏颇体质的人,更容易出现亚健康、疾病状态。由此我们推测,气虚质与健康状态有着密切的联系。健康人亦存在气虚质,且对健康气虚质人群进行研究排除了疾病对监测结果的干扰,因此,我们设计了对健康人气虚质进行中药干预的随机、对照研究,同时应用代谢组学进行气虚质代谢紊乱的机制研究。
代谢组学(metabonomics)是通过组群指标分析,进行高通量检测和数据处理,研究生物体整体或组织细胞系统的动态代谢变化,特别是对内源代谢、遗传变异、环境变化乃至各种物质进入代谢系统的特征和影响的学科。代谢组学是对新陈代谢过程中所有低分子量(<1kD)代谢产物进行定性和定量的研究,以反映生物体对外界刺激或基因修饰所发生的变化的学科。与转录组学、蛋白组学相比,代谢组学有自身的优点。首先,代谢组是最接近表型,最能反应表型变化的,基因表达的变化引起蛋白表达的变化,最终以代谢过程的变化发生效应。因此,代谢组学研究在了解事物本质变化中具有重要的作用,基因和蛋白表达的微小变化还会在代谢物上得到放大。而且代谢物的数量比基因和蛋白的数量少得多。因此,我们选择代谢组学对气虚质的代谢紊乱进行研究。本课题组对亚健康气虚质的代谢组学研究发现,其尿液中肌酸酐、组胺、葡萄糖、丙氨酸、鸟苷、三磷酸甘油酸、磷酸肌酸、马尿酸、组氨酸、甘油酸水平显著升高,这些代谢产物与三羧酸循环、糖酵解、氨基酸代谢、脂肪酸代谢等多个能量代谢通路相关。由前期研究结果可见,长期繁重的工作和精神紧张导致机体气血亏虚,能量消耗加快,代谢产物增多,使人产生疲乏感,出现亚健康状态。而健康的气虚质人,其体内阴阳气血已经失调,更容易受外界环境等因素影响导致亚健康、疾病的发生。因此,我们推测,气虚质存在代谢紊乱,给予益气中药干预可以调整气虚质的这种代谢紊乱,预防亚健康、疾病的发生。
补中益气丸出自《内外伤辨惑论》,是金元四大家之一的“补土派”代表李东垣的代表方,主治气虚发热、泄泻,气虚不能固摄血液、下陷等症状。现代药理学研究发现补中益气汤具有调节胃肠运动、影响消化液分泌、抗肿瘤、抗突变、调节免疫力、抑菌消炎等综合作用。因此,补中益气丸很适合调理以气短懒言,容易疲乏,精神不振,易出汗等为主要表现,并伴有脾胃消化不良等不适症状的气虚质人群。
目的：
1、明确补中益气丸干预健康气虚质人群的临床疗效,为应用中医药手段干预中医偏颇体质提供科学依据。
2、探讨补中益气丸干预健康气虚质人群代谢紊乱的机制。
方法：1、进行健康人气虚质中药干预的研究。应用中医体质量表、亚健康调查表、生活方式量表和体检报告进行健康与体质状况判断,再进行面谈并采用中医四诊合参方法,进一步筛选健康人单纯气虚质,并将其随机分为气虚质对照组和药物干预组,干预组应用补中益气丸干预1个月,对照组不进行干预。随访观察研究对象的主要疗效指标为气虚质转化分改变情况,次要疗效指标为其他体质转化分改变情况和亚健康量表评分和生活方式量表评分,通过统计分析主要及次要疗效指标,以明确应用中药调理气虚质的临床疗效,为应用中医药手段干预中医偏颇体质提供科学依据。
2、进行平和质与气虚质的代谢组学研究。从论文第二章临床观察的人群中筛选出气虚质,并纳入平和质人群,清晨空腹抽血后提取血浆-20℃冻存。血浆进行代谢组学特征分析后,对其结果进行生物信息学分析,建立代谢组差异代谢图谱库。
3、进行补中益气丸干预气虚质的代谢组学研究。从论文第二章临床观察的人群中筛选出气虚质人群,对其用药前后干预组与对照组人群,清晨空腹抽血后提取血浆-20℃冻存。血浆进行代谢组学特征分析后,对其结果进行生物信息学分析,建立代谢组差异代谢图谱库。
4、汇总代谢组学数据进行分析,采用MestReNova5.3软件进行代谢图谱预处理,对各分段积分值进行归一化处理。所得数据输出并转换到Excel格式并导入软件Simca-P12.0.1.0(Umetrics AB, Sweden)中进行多元统计分析。采用主成分分析(Principal Component Analysis, PC A)和偏最小二乘法判别法(Partial least-squares discriminant analysis, PLS-DA)进行分析,为强化组间差异,进一步采用正交偏最小二乘判别分析(OPLS-DA)。分析结果以得分图(Scores plot)的形式表示。统计分析软件采用SPSS13.0。对血浆中的几种代谢物的归一化积分值(X±S表示)做统计学处理(两独立样本t检验)。在PLS-DA模型中提取VIP值(variable importance in projection, VIP),选取VIP值>1的化学位移值,进一步对这些化学位移值的归一化积分进行统计学分析,提取P<0.05的化学位移值进行差异代谢产物成分鉴定。通过人类代谢产物数据库(http://www.hmdb.ca)及查阅文献来鉴定其代谢产物名称,初步探讨补中益气丸干预气虚质代谢紊乱的机制。
结果：
1、补中益气丸干预气虚质的临床观察研究
通过初步调查、面谈筛选出愿意入组的健康气虚质研究对象74例,其中6例在研究过程中退出,最后完成观察共68例(对照组和干预组各34例)。分析结果：①用药前,气虚质对照组与干预组之间的主要评价指标和次要评价指标均无统计学差异(P>0.05)。②干预1个月后,主要评价指标即气虚质转化分,干预组有所下降(分数越高气虚质症状越明显),其用药前后的差异有统计学意义(t=4.355,P=0.000),而对照组在观察1个月后气虚质转化分较观察前差异无统计学意义(t=-1.145,P=0.26)。干预1个月后,对照组与干预组之间气虚质转化分差异有统计学意义(t=2.053,P=0.044)。③在次要评价指标中,干预1个月后,干预组用药前后中医体质量表中其他体质分数的变化无统计学意义(P>0.05)；而对照组在观察1个月后,其阳虚质、阴虚质、血瘀质分数较观察前有所升高(分数越高偏颇体质症状越明显),差异有统计学意义(P<0.05)。④干预1个月后,干预组生活方式评分有所升高(分数越高生活方式越健康),差异有统计学意义(t=-2.879,P=0.007)。而对照组分数变化无统计学意义(t=-1.366,P=0.181)。干预组与对照组亚健康评分无统计学差异(t=-0.811,P=0.421)。⑤在治疗前后气虚质量表各条目的比较中发现,干预组心慌、头晕、懒言症状分数有所下降,差异有统计学意义(P<0.05),对照组1个月后症状分数有所变化,但差异无统计学意义(P>0.05)。
2、健康气虚质与健康平和质血浆1H NMR分析
从论文第二章临床观察的气虚质人群中筛选出气虚质19人,并纳入平和质15人,采集清晨空腹血液样本进行代谢组学检测后,应用分析软件对图谱进行分析,采用PCA方法对气虚质与平和质样本建模,累计计算出6个主成分,模型参数R2X=0.856,Q2=0.504,模型分析了85.6%的原始数据。基于气虚质与平和质血浆代谢数据建立OPLS-DA模型,对两组样本建模,累计计算出2个主成分,模型参数R2X=0.531, R2Y=0.388, Q2=0.0787。可见气虚质与平和质之间可明确区分。气虚质组与平和质组血浆中对分类贡献较大的特异性代谢物11个,包括甜菜碱、脯氨酸、亮氨酸、谷氨酰胺、丝氨酸、组氨酸、丙氨酸、烟酰胺核苷酸、异亮氨酸、门冬氨酸、肌醇。这些差异代谢物与脂肪酸代谢、三羧酸循环、鸟氨酸循环、氨基酸代谢、丙酮酸代谢、糖异生、丙氨酸循环、肌醇代谢、嘌呤嘧啶代谢、氨回收、神经递质传导、甘氨酸和丝氨酸代谢等多个代谢通路相关。
3、补中益气丸干预组用药前后血浆代谢组学的研究
从论文第二章临床观察的气虚质人群中筛选出气虚质20人(对照组和干预组各10人),采集干预组用药前、用药后和气虚质对照组志愿者血液(干预组有1人用药前标本因样本问题未能检测)进行代谢组学检测后,应用分析软件对图谱进行分析,采用PCA方法对干预组用药前后样本建模,累计计算出5个主成分,模型参数R2X=0.725, Q2X=0.382,模型分析了72.5%的原始数据。基于补中益气丸用药前后干预组的血浆代谢数据建立PLS-DA模型,对两组样本建模,累计计算出3个主成分,模型参数R2X=0.502, R2Y=0.967, Q2X=0.721。可见干预组在药物干预前后的样本可显著区分。干预组用药前后血浆中对分类贡献较大的特异性代谢物35个,包括肾固醇、辅酶A、谷固醇、醛固酮、缬氨酸、胆固醇硫酸盐、乙醇、前列腺素El、异亮氨酸、丙氨酸、γ-氨基丁酸、谷氨酰胺、蛋氨酸、甜菜碱、苯丙氨酸、苯丙醇胺、丙氨酸、甘油酸、天冬氨酸、丝氨酸、组氨酸、α-亚麻酸、前列腺素D2、2-乙基丙烯酸、异维A酸、鞘氨醇、尿素、酪氨酸、’肾上腺素、乙酰氨基、甲基马尿酸、香草酸、马尿酸、N-乙酰基酪氨酸、羟基苯甲酸。多为糖、脂肪、蛋白质三大营养能量代谢过程的中间产物,与脂肪酸代谢、三羧酸循环、鸟氨酸循环、儿茶酚胺合成、氨基酸代谢、丙酮酸代谢、糖异生、类固醇合成、嘌呤嘧啶代谢、鞘脂类代谢、视黄醇的代谢、氨回收、磷脂合成代谢、甘氨酸和丝氨酸代谢、神经递质传导、线粒体电子传递链等多个代谢通路相关。
结论：1、气虚质对照组观察1个月后发现其有向其他体质转化变成复合体质的倾向。
2、气虚质可以通过补中益气丸进行改善和调理。
3、应用补中益气丸干预气虚质后,其与三羧酸循环、氨合成相关的氨基酸、脂类、蛋白质有所降低,说明补中益气丸调节了气虚质的这种代谢紊乱。
Backgroud:Traditional Chinese Medicine (TCM) constitution is the relatively stable, inherent characteristics formed on the basis of innate endowment and acquired characteristics after birth, integrated with morphological structure, physiological function and psychological status in the process of human life. The qi-deficiency is characteristic as low voice, shortness of breath, lassitude, easy to sweat, weak pulse, reddish tongue with tooth print. So far, the study on qi-deficiency is focus on epidemiological investigation and poor in genes, proteins and metabolic mechanism. Our previous studies suggested that the number of bias constitution in sub-health status is more than that in health. As a result, we inferred that the qi-deficiency constitution is associated with health status. In this study, a randomized, controlled study was conducted to study the bu zhong yi qi wan intervening qi-deficiency constitution to test its mechanism.
The study on qi-deficiency still aimed at epidemiological investigation and poor in gene chip, proteomics, metabonomic. Our previous result show:(Dthe number of bias constitution in health status is in minority (18.7%), and the gentleness constitution in health status is in majority (81.3%). The number of bias constitution in sub-health status is more than that in health (64.0%), most of whom are qi-deficiency constitution (41%).②As the logistic result shows, the gentleness constitution is positive related to health status and negative related to sub-health status. The qi-deficiency constitution is positive related to sub-health status.③The gentleness constitution is hard to transforme into sub-health status and disease, while the bias constitution is much more easier in sub-health status and disease. As a result, we inferred that the bias constitution is associated with health status. The qi-deficiency constitution volunteers in health status cannot be interferenced by disease. Therefore, we studied the metabonomic characteristics of qi-deficiency constitution volunteers in health status, and intervented them by bu zhong yi qi wan to study its mechanism.
Metabolomics is processing by group index analysis, high-throughput detection and data. It investigated the overall dynamic metabolic changes, particularly in studing the characteristics and effects of endogenous metabolic, genetic variation, environment changes, as well as a variety of substances transferred into metabolic systems, of. Metabolomics is qualitative and quantitative analysis low molecular metabolites (<1kD) to reflect changes in the organism after external stimuli or genetic modification. Compared with transcriptomics and proteomics, metabolomics has its own advantages. Firstly, the metabolic is the closest to the phenotype, which can reflect phenotypic changes. The changes in gene caused the changes of protein expression, and ultimately occured changes in metabolic processes. Therefore, the study of the metabolic group has an important role in understanding the changes of nature. Small changes in gene and protein expression will be amplified in the metabolite. The number of metabolites is much less than the number of genes and proteins. As a result, we used the metabolitic to study the metabolic disorders of qi-deficiency constitution. Our group found that the urine of qi-deficiency constitution contained creatinine, histamine, glucose, alanine, guanosine triphosphate glycerol acid, creatine phosphate, hippuric acid, histidine, glycerol acid levels. These metabolites, which were related with multiple energy krebs cycle, glycolysis, amino acid metabolism, fatty acid metabolism and other metabolic pathway, were significantly increased. The results from the preliminary study shows that long-term heavy work and mental stress causes the body and blood problems, the energy consumption rate increased, gives rise to a sense of fatigue, and at last in sub-health status. The qi-deficiency constitution participates have yin and yang imbalance, more susceptible to the external environment and other factors lead to sub-health or disease. Therefore, we hypothesized that the presence of energy metabolism in qi-deficiency constitution, may contribute to the health sub-health or disease. The intervention of qi-deficiency constitution can be adjusted to prevent the occurrence of sub-health and disease.
Bu zhong yi qi wan is the representative prescription for Li Dongheng, who is one of the four famous doctors in Jinyuan period. The medicine can treat fever, diarrhea, bleeding and others. Modern pharmacology studies found that the element of bu zhong yi qi wan is complexity, which contains saponins, polysaccharides, flavonoids, alkaloids, amino acids and volatile oil. It is useful to regulate gastrointestinal motility, secretion of digestive juice, promote intestinal absorption, anti-tumor, regulate immune function, promoting protein synthesis, anoxia, and combined effects of antibacterial anti-inflammatory. The qi-deficiency is characteristic as low voice, shortness of breath, lassitude, easy to sweat, weak pulse, reddish tongue with tooth print and usually have some digest problems. The bu zhong yi qi wan is suitable for regulate qi-deficiency.
Object:Clarify the curative effect of bu zhong yi qi wan intervening qi-deficiency constitution. Clarify the metabolic disorders of bu zhong yi qi wan intervening qi-deficiency constitution
Method:Firstly, we study the TCM intervening qi-deficiency constitution. The sub-health questionnaire and physical examination have been used to determine the health status. We selected qi-deficiency constitution in health status and randomly divided them into control group and intervention group. The intervention group was intervened by Bu zhong yi qi wan for a month and the control group was not intervened. The main curative index is converted scores of qi-deficency subscale and the subordination curative index is converted scores of CCMQ, SHMS V1.0and HPLP-II. The health status has been observed to clarify the effect of TCM on health transformation.
Secondly, we study the metabolomics of qi-deficiency constitution and gentleness constitution. We selected19qi-deficiency constitution people and15gentleness constitution. The plasma was collected in empty stomach and cryopreserved at-20℃.
Thirdly, we study the metabolomics of qi-deficency intervention group before and after intervention. We selected20qi-deficiency constitution people (10in control group and10in intervention group). The plasma was collected in empty stomach and cryopreserved at-20℃.
Fourthly, analysed the metabolomics data. The plasma was thawed at room temperature and centrifuged (14000×g,10min,4＞).0.3ml supernatant buffer solution,0.2ml PBS,0.1ml heavy water was added together, and injected in5mm NMR tubes to test metabolic genomics. The plasma of intervention group, control group and gentleness group had been collected before and after intervention for the metabolomics analysis. MestReNova5.3software has been used to pretreatment the metabolic profiles, and normalized the values for each segment integral. The data output will be converted to Excel format and imported into Simca-P12.0.1.0(Umetrics AB, Sweden) for multivariate statistical analysis. Principal component analysis and partial least squares discriminant analysis have been used to analyse. In order to strengthen inter-group differences, the orthogonal partial least squares discriminant analysis (OPLS-DA) has been used as well. The results were expressed in scores plot. Statistical analysis was performed using SPSS13.0. The normalization value (X±S) of the plasma metabolites has analysis by two independent samples t test. VIP values have been extracted from PLS-DA model. We selected metabolites VIP value>1, P<0.05as difference variables. Combination of clinical symptoms, medical reports and questionnaires content, VIP value and normalized results were identified to find represent metabolites. The component identification was identified through human metabolite database (http://www.hmdb.ca) and literature. We found related metabolic markers of qi-deficiency constitution to explore mechanisms of energy metabolism disorder.
Results:
(1) Clinical observation of TCM intervening qi-deficiency constitution
Before the intervention, there are no significantly different between control group and intervention group. After a month later, the scores of qi-deficiency constitution in intervention group have decreased significantly (t=4.355, P=0.000), while the control group was increased, but there was no significant difference (t=-1.145, P=0.26). The control group and intervention group had significant difference after the intervention (t=2.053, P=0.044). Bu zhong yi qi wan can improve the symptoms significantly. In the secondary evaluation indicators, scores of other constitutions don't change after a month intervention in intervention group (P>0.05), while the scores of yang-deficiency constitution, yin-deficiency constitution, blood stasis constitution changed in the control group, the difference was statistically significant (P<0.05). After a month later, the scores of HPLP-II in intervention group have decreased significantly (t=-2.879, P=0.007), but there was no significant difference in control group(t=-1.366, P=0.181). There are no significant difference in control group and intervention group in HPLP-II after1month (t=-0.811, P=0.421). The scores of some symptom increased in control group after1month, but there was no significant difference, while in intervention group, the scores have declined. The scores of palpitation, dizziness, lazy improved significantly (P<0.05).
(2) The plasma1H NMR profiling of qi-deficiency constitution and gentleness constitution
We selected35volenteers (15in gentleness constitution and19in qi-deficency constitution) to study its metabolism characteristics. PCA method was used to build model and six main components was calculated. The model parameters R2X=0.856, Q2=0.504, the model analyzed85.6%of the original data. Based on the data to establish the OPLS-DA model and two main components was calculated. The model parameters R2X=0.531, R2Y=0.388, Q2=0.0787. There are significantly differences between the two samples after intervention. There are11specific metabolites, including betaine, proline, leucine, glutamine, serine, histidine, alanine, nicotinamide nucleotide, isoleucine, aspartic acid, inositol. It is related to fatty acid metabolism, Krebs cycle, ornithine cycle, amino acid metabolism, pyruvate metabolism, gluconeogenesis, alanine cycle, inositol metabolism, steroid synthesis, purine pyrimidine metabolism, ammonia recovery, glycine and serine metabolism multiple metabolic pathway.
(3) The plasma1H NMR profiling of intervention group before and after intervention
We selected20volenteers (10in control group and10in intervention group) to study its metabolism characteristics. PCA method was used to build model and five main components was calculated. The model parameters R2X=0.725, Q2X=0.382, the model analyzed72.5%of the original data. Based on the data to establish the PLS-DA model and three main components was calculated. The model parameters R2X=0.502, R2Y=0.967, Q2X=0.721. There are significantly differences in intervention group before and after intervention. There are35specific metabolites, including renal steroids, coenzyme A, sitosterol, aldosterone, valine, cholesterol sulfate, ethanol, prostaglandin E1, isoleucine, alanine, gamma amino acid, glutamine, methionine, betaine, benzene alanine, phenylpropanolamine, alanine, glycerol acid, aspartic acid, serine, histidine, a-linolenic acid, prostaglandin D2,2-ethyl acrylate, different dimension A acid, sphingosine, urea, tyrosine, epinephrine, acetylamino, methyl hippuric acid, vanillic acid, hippuric acid, N-acetyl-tyrosine, p-hydroxybenzoic acid. It is related to fatty acid metabolism, the TCA cycle, ornithine cycle, catecholamine synthesis, amino acid metabolism, pyruvate metabolism, gluconeogenesis, synthesis of steroids, purine pyrimidine metabolism, metabolism of sphingolipids metabolism, retinol, ammonia recovery, synthetic phospholipids metabolism, glycine and serine metabolism, mitochondrial electron transport chain and multiple metabolic pathways.
Conclusion:Firstly, it indirectly shows that if the constitution can not be improved, it may convert to other constitutions.
Secondly, Bu zhong yi qi wan can significantly improve the qi-deficiency constitution.
Thirdly, the different metabolites of intervention group before and after intervention related to the citric acid cycle and bu zhong yi qi wan maybe regulate the disorder.
目前对气虚质的研究仍处于流行病学调查阶段,其基因、蛋白、代谢方面的机制研究并不多。根据本课题组前期对体质的研究结果发现：①广东地区健康人群以平和质居多(占81.3%),偏颇体质仅占18.7%；亚健康人群以偏颇体质居多(占64.0%),其中气虚质所占比例最大(占41%)。②对健康、亚健康人群的中医体质进行Logistic回归发现,平和质与健康呈正相关,与亚健康呈负相关,气虚质与亚健康呈正相关。③平和质的人,不容易出现亚健康、疾病；偏颇体质的人,更容易出现亚健康、疾病状态。由此我们推测,气虚质与健康状态有着密切的联系。健康人亦存在气虚质,且对健康气虚质人群进行研究排除了疾病对监测结果的干扰,因此,我们设计了对健康人气虚质进行中药干预的随机、对照研究,同时应用代谢组学进行气虚质代谢紊乱的机制研究。
代谢组学(metabonomics)是通过组群指标分析,进行高通量检测和数据处理,研究生物体整体或组织细胞系统的动态代谢变化,特别是对内源代谢、遗传变异、环境变化乃至各种物质进入代谢系统的特征和影响的学科。代谢组学是对新陈代谢过程中所有低分子量(<1kD)代谢产物进行定性和定量的研究,以反映生物体对外界刺激或基因修饰所发生的变化的学科。与转录组学、蛋白组学相比,代谢组学有自身的优点。首先,代谢组是最接近表型,最能反应表型变化的,基因表达的变化引起蛋白表达的变化,最终以代谢过程的变化发生效应。因此,代谢组学研究在了解事物本质变化中具有重要的作用,基因和蛋白表达的微小变化还会在代谢物上得到放大。而且代谢物的数量比基因和蛋白的数量少得多。因此,我们选择代谢组学对气虚质的代谢紊乱进行研究。本课题组对亚健康气虚质的代谢组学研究发现,其尿液中肌酸酐、组胺、葡萄糖、丙氨酸、鸟苷、三磷酸甘油酸、磷酸肌酸、马尿酸、组氨酸、甘油酸水平显著升高,这些代谢产物与三羧酸循环、糖酵解、氨基酸代谢、脂肪酸代谢等多个能量代谢通路相关。由前期研究结果可见,长期繁重的工作和精神紧张导致机体气血亏虚,能量消耗加快,代谢产物增多,使人产生疲乏感,出现亚健康状态。而健康的气虚质人,其体内阴阳气血已经失调,更容易受外界环境等因素影响导致亚健康、疾病的发生。因此,我们推测,气虚质存在代谢紊乱,给予益气中药干预可以调整气虚质的这种代谢紊乱,预防亚健康、疾病的发生。
补中益气丸出自《内外伤辨惑论》,是金元四大家之一的“补土派”代表李东垣的代表方,主治气虚发热、泄泻,气虚不能固摄血液、下陷等症状。现代药理学研究发现补中益气汤具有调节胃肠运动、影响消化液分泌、抗肿瘤、抗突变、调节免疫力、抑菌消炎等综合作用。因此,补中益气丸很适合调理以气短懒言,容易疲乏,精神不振,易出汗等为主要表现,并伴有脾胃消化不良等不适症状的气虚质人群。
目的：
1、明确补中益气丸干预健康气虚质人群的临床疗效,为应用中医药手段干预中医偏颇体质提供科学依据。
2、探讨补中益气丸干预健康气虚质人群代谢紊乱的机制。
方法：1、进行健康人气虚质中药干预的研究。应用中医体质量表、亚健康调查表、生活方式量表和体检报告进行健康与体质状况判断,再进行面谈并采用中医四诊合参方法,进一步筛选健康人单纯气虚质,并将其随机分为气虚质对照组和药物干预组,干预组应用补中益气丸干预1个月,对照组不进行干预。随访观察研究对象的主要疗效指标为气虚质转化分改变情况,次要疗效指标为其他体质转化分改变情况和亚健康量表评分和生活方式量表评分,通过统计分析主要及次要疗效指标,以明确应用中药调理气虚质的临床疗效,为应用中医药手段干预中医偏颇体质提供科学依据。
2、进行平和质与气虚质的代谢组学研究。从论文第二章临床观察的人群中筛选出气虚质,并纳入平和质人群,清晨空腹抽血后提取血浆-20℃冻存。血浆进行代谢组学特征分析后,对其结果进行生物信息学分析,建立代谢组差异代谢图谱库。
3、进行补中益气丸干预气虚质的代谢组学研究。从论文第二章临床观察的人群中筛选出气虚质人群,对其用药前后干预组与对照组人群,清晨空腹抽血后提取血浆-20℃冻存。血浆进行代谢组学特征分析后,对其结果进行生物信息学分析,建立代谢组差异代谢图谱库。
4、汇总代谢组学数据进行分析,采用MestReNova5.3软件进行代谢图谱预处理,对各分段积分值进行归一化处理。所得数据输出并转换到Excel格式并导入软件Simca-P12.0.1.0(Umetrics AB, Sweden)中进行多元统计分析。采用主成分分析(Principal Component Analysis, PC A)和偏最小二乘法判别法(Partial least-squares discriminant analysis, PLS-DA)进行分析,为强化组间差异,进一步采用正交偏最小二乘判别分析(OPLS-DA)。分析结果以得分图(Scores plot)的形式表示。统计分析软件采用SPSS13.0。对血浆中的几种代谢物的归一化积分值(X±S表示)做统计学处理(两独立样本t检验)。在PLS-DA模型中提取VIP值(variable importance in projection, VIP),选取VIP值>1的化学位移值,进一步对这些化学位移值的归一化积分进行统计学分析,提取P<0.05的化学位移值进行差异代谢产物成分鉴定。通过人类代谢产物数据库(http://www.hmdb.ca)及查阅文献来鉴定其代谢产物名称,初步探讨补中益气丸干预气虚质代谢紊乱的机制。
结果：
1、补中益气丸干预气虚质的临床观察研究
通过初步调查、面谈筛选出愿意入组的健康气虚质研究对象74例,其中6例在研究过程中退出,最后完成观察共68例(对照组和干预组各34例)。分析结果：①用药前,气虚质对照组与干预组之间的主要评价指标和次要评价指标均无统计学差异(P>0.05)。②干预1个月后,主要评价指标即气虚质转化分,干预组有所下降(分数越高气虚质症状越明显),其用药前后的差异有统计学意义(t=4.355,P=0.000),而对照组在观察1个月后气虚质转化分较观察前差异无统计学意义(t=-1.145,P=0.26)。干预1个月后,对照组与干预组之间气虚质转化分差异有统计学意义(t=2.053,P=0.044)。③在次要评价指标中,干预1个月后,干预组用药前后中医体质量表中其他体质分数的变化无统计学意义(P>0.05)；而对照组在观察1个月后,其阳虚质、阴虚质、血瘀质分数较观察前有所升高(分数越高偏颇体质症状越明显),差异有统计学意义(P<0.05)。④干预1个月后,干预组生活方式评分有所升高(分数越高生活方式越健康),差异有统计学意义(t=-2.879,P=0.007)。而对照组分数变化无统计学意义(t=-1.366,P=0.181)。干预组与对照组亚健康评分无统计学差异(t=-0.811,P=0.421)。⑤在治疗前后气虚质量表各条目的比较中发现,干预组心慌、头晕、懒言症状分数有所下降,差异有统计学意义(P<0.05),对照组1个月后症状分数有所变化,但差异无统计学意义(P>0.05)。
2、健康气虚质与健康平和质血浆1H NMR分析
从论文第二章临床观察的气虚质人群中筛选出气虚质19人,并纳入平和质15人,采集清晨空腹血液样本进行代谢组学检测后,应用分析软件对图谱进行分析,采用PCA方法对气虚质与平和质样本建模,累计计算出6个主成分,模型参数R2X=0.856,Q2=0.504,模型分析了85.6%的原始数据。基于气虚质与平和质血浆代谢数据建立OPLS-DA模型,对两组样本建模,累计计算出2个主成分,模型参数R2X=0.531, R2Y=0.388, Q2=0.0787。可见气虚质与平和质之间可明确区分。气虚质组与平和质组血浆中对分类贡献较大的特异性代谢物11个,包括甜菜碱、脯氨酸、亮氨酸、谷氨酰胺、丝氨酸、组氨酸、丙氨酸、烟酰胺核苷酸、异亮氨酸、门冬氨酸、肌醇。这些差异代谢物与脂肪酸代谢、三羧酸循环、鸟氨酸循环、氨基酸代谢、丙酮酸代谢、糖异生、丙氨酸循环、肌醇代谢、嘌呤嘧啶代谢、氨回收、神经递质传导、甘氨酸和丝氨酸代谢等多个代谢通路相关。
3、补中益气丸干预组用药前后血浆代谢组学的研究
从论文第二章临床观察的气虚质人群中筛选出气虚质20人(对照组和干预组各10人),采集干预组用药前、用药后和气虚质对照组志愿者血液(干预组有1人用药前标本因样本问题未能检测)进行代谢组学检测后,应用分析软件对图谱进行分析,采用PCA方法对干预组用药前后样本建模,累计计算出5个主成分,模型参数R2X=0.725, Q2X=0.382,模型分析了72.5%的原始数据。基于补中益气丸用药前后干预组的血浆代谢数据建立PLS-DA模型,对两组样本建模,累计计算出3个主成分,模型参数R2X=0.502, R2Y=0.967, Q2X=0.721。可见干预组在药物干预前后的样本可显著区分。干预组用药前后血浆中对分类贡献较大的特异性代谢物35个,包括肾固醇、辅酶A、谷固醇、醛固酮、缬氨酸、胆固醇硫酸盐、乙醇、前列腺素El、异亮氨酸、丙氨酸、γ-氨基丁酸、谷氨酰胺、蛋氨酸、甜菜碱、苯丙氨酸、苯丙醇胺、丙氨酸、甘油酸、天冬氨酸、丝氨酸、组氨酸、α-亚麻酸、前列腺素D2、2-乙基丙烯酸、异维A酸、鞘氨醇、尿素、酪氨酸、’肾上腺素、乙酰氨基、甲基马尿酸、香草酸、马尿酸、N-乙酰基酪氨酸、羟基苯甲酸。多为糖、脂肪、蛋白质三大营养能量代谢过程的中间产物,与脂肪酸代谢、三羧酸循环、鸟氨酸循环、儿茶酚胺合成、氨基酸代谢、丙酮酸代谢、糖异生、类固醇合成、嘌呤嘧啶代谢、鞘脂类代谢、视黄醇的代谢、氨回收、磷脂合成代谢、甘氨酸和丝氨酸代谢、神经递质传导、线粒体电子传递链等多个代谢通路相关。
结论：1、气虚质对照组观察1个月后发现其有向其他体质转化变成复合体质的倾向。
2、气虚质可以通过补中益气丸进行改善和调理。
3、应用补中益气丸干预气虚质后,其与三羧酸循环、氨合成相关的氨基酸、脂类、蛋白质有所降低,说明补中益气丸调节了气虚质的这种代谢紊乱。
Backgroud:Traditional Chinese Medicine (TCM) constitution is the relatively stable, inherent characteristics formed on the basis of innate endowment and acquired characteristics after birth, integrated with morphological structure, physiological function and psychological status in the process of human life. The qi-deficiency is characteristic as low voice, shortness of breath, lassitude, easy to sweat, weak pulse, reddish tongue with tooth print. So far, the study on qi-deficiency is focus on epidemiological investigation and poor in genes, proteins and metabolic mechanism. Our previous studies suggested that the number of bias constitution in sub-health status is more than that in health. As a result, we inferred that the qi-deficiency constitution is associated with health status. In this study, a randomized, controlled study was conducted to study the bu zhong yi qi wan intervening qi-deficiency constitution to test its mechanism.
The study on qi-deficiency still aimed at epidemiological investigation and poor in gene chip, proteomics, metabonomic. Our previous result show:(Dthe number of bias constitution in health status is in minority (18.7%), and the gentleness constitution in health status is in majority (81.3%). The number of bias constitution in sub-health status is more than that in health (64.0%), most of whom are qi-deficiency constitution (41%).②As the logistic result shows, the gentleness constitution is positive related to health status and negative related to sub-health status. The qi-deficiency constitution is positive related to sub-health status.③The gentleness constitution is hard to transforme into sub-health status and disease, while the bias constitution is much more easier in sub-health status and disease. As a result, we inferred that the bias constitution is associated with health status. The qi-deficiency constitution volunteers in health status cannot be interferenced by disease. Therefore, we studied the metabonomic characteristics of qi-deficiency constitution volunteers in health status, and intervented them by bu zhong yi qi wan to study its mechanism.
Metabolomics is processing by group index analysis, high-throughput detection and data. It investigated the overall dynamic metabolic changes, particularly in studing the characteristics and effects of endogenous metabolic, genetic variation, environment changes, as well as a variety of substances transferred into metabolic systems, of. Metabolomics is qualitative and quantitative analysis low molecular metabolites (<1kD) to reflect changes in the organism after external stimuli or genetic modification. Compared with transcriptomics and proteomics, metabolomics has its own advantages. Firstly, the metabolic is the closest to the phenotype, which can reflect phenotypic changes. The changes in gene caused the changes of protein expression, and ultimately occured changes in metabolic processes. Therefore, the study of the metabolic group has an important role in understanding the changes of nature. Small changes in gene and protein expression will be amplified in the metabolite. The number of metabolites is much less than the number of genes and proteins. As a result, we used the metabolitic to study the metabolic disorders of qi-deficiency constitution. Our group found that the urine of qi-deficiency constitution contained creatinine, histamine, glucose, alanine, guanosine triphosphate glycerol acid, creatine phosphate, hippuric acid, histidine, glycerol acid levels. These metabolites, which were related with multiple energy krebs cycle, glycolysis, amino acid metabolism, fatty acid metabolism and other metabolic pathway, were significantly increased. The results from the preliminary study shows that long-term heavy work and mental stress causes the body and blood problems, the energy consumption rate increased, gives rise to a sense of fatigue, and at last in sub-health status. The qi-deficiency constitution participates have yin and yang imbalance, more susceptible to the external environment and other factors lead to sub-health or disease. Therefore, we hypothesized that the presence of energy metabolism in qi-deficiency constitution, may contribute to the health sub-health or disease. The intervention of qi-deficiency constitution can be adjusted to prevent the occurrence of sub-health and disease.
Bu zhong yi qi wan is the representative prescription for Li Dongheng, who is one of the four famous doctors in Jinyuan period. The medicine can treat fever, diarrhea, bleeding and others. Modern pharmacology studies found that the element of bu zhong yi qi wan is complexity, which contains saponins, polysaccharides, flavonoids, alkaloids, amino acids and volatile oil. It is useful to regulate gastrointestinal motility, secretion of digestive juice, promote intestinal absorption, anti-tumor, regulate immune function, promoting protein synthesis, anoxia, and combined effects of antibacterial anti-inflammatory. The qi-deficiency is characteristic as low voice, shortness of breath, lassitude, easy to sweat, weak pulse, reddish tongue with tooth print and usually have some digest problems. The bu zhong yi qi wan is suitable for regulate qi-deficiency.
Object:Clarify the curative effect of bu zhong yi qi wan intervening qi-deficiency constitution. Clarify the metabolic disorders of bu zhong yi qi wan intervening qi-deficiency constitution
Method:Firstly, we study the TCM intervening qi-deficiency constitution. The sub-health questionnaire and physical examination have been used to determine the health status. We selected qi-deficiency constitution in health status and randomly divided them into control group and intervention group. The intervention group was intervened by Bu zhong yi qi wan for a month and the control group was not intervened. The main curative index is converted scores of qi-deficency subscale and the subordination curative index is converted scores of CCMQ, SHMS V1.0and HPLP-II. The health status has been observed to clarify the effect of TCM on health transformation.
Secondly, we study the metabolomics of qi-deficiency constitution and gentleness constitution. We selected19qi-deficiency constitution people and15gentleness constitution. The plasma was collected in empty stomach and cryopreserved at-20℃.
Thirdly, we study the metabolomics of qi-deficency intervention group before and after intervention. We selected20qi-deficiency constitution people (10in control group and10in intervention group). The plasma was collected in empty stomach and cryopreserved at-20℃.
Fourthly, analysed the metabolomics data. The plasma was thawed at room temperature and centrifuged (14000×g,10min,4＞).0.3ml supernatant buffer solution,0.2ml PBS,0.1ml heavy water was added together, and injected in5mm NMR tubes to test metabolic genomics. The plasma of intervention group, control group and gentleness group had been collected before and after intervention for the metabolomics analysis. MestReNova5.3software has been used to pretreatment the metabolic profiles, and normalized the values for each segment integral. The data output will be converted to Excel format and imported into Simca-P12.0.1.0(Umetrics AB, Sweden) for multivariate statistical analysis. Principal component analysis and partial least squares discriminant analysis have been used to analyse. In order to strengthen inter-group differences, the orthogonal partial least squares discriminant analysis (OPLS-DA) has been used as well. The results were expressed in scores plot. Statistical analysis was performed using SPSS13.0. The normalization value (X±S) of the plasma metabolites has analysis by two independent samples t test. VIP values have been extracted from PLS-DA model. We selected metabolites VIP value>1, P<0.05as difference variables. Combination of clinical symptoms, medical reports and questionnaires content, VIP value and normalized results were identified to find represent metabolites. The component identification was identified through human metabolite database (http://www.hmdb.ca) and literature. We found related metabolic markers of qi-deficiency constitution to explore mechanisms of energy metabolism disorder.
Results:
(1) Clinical observation of TCM intervening qi-deficiency constitution
Before the intervention, there are no significantly different between control group and intervention group. After a month later, the scores of qi-deficiency constitution in intervention group have decreased significantly (t=4.355, P=0.000), while the control group was increased, but there was no significant difference (t=-1.145, P=0.26). The control group and intervention group had significant difference after the intervention (t=2.053, P=0.044). Bu zhong yi qi wan can improve the symptoms significantly. In the secondary evaluation indicators, scores of other constitutions don't change after a month intervention in intervention group (P>0.05), while the scores of yang-deficiency constitution, yin-deficiency constitution, blood stasis constitution changed in the control group, the difference was statistically significant (P<0.05). After a month later, the scores of HPLP-II in intervention group have decreased significantly (t=-2.879, P=0.007), but there was no significant difference in control group(t=-1.366, P=0.181). There are no significant difference in control group and intervention group in HPLP-II after1month (t=-0.811, P=0.421). The scores of some symptom increased in control group after1month, but there was no significant difference, while in intervention group, the scores have declined. The scores of palpitation, dizziness, lazy improved significantly (P<0.05).
(2) The plasma1H NMR profiling of qi-deficiency constitution and gentleness constitution
We selected35volenteers (15in gentleness constitution and19in qi-deficency constitution) to study its metabolism characteristics. PCA method was used to build model and six main components was calculated. The model parameters R2X=0.856, Q2=0.504, the model analyzed85.6%of the original data. Based on the data to establish the OPLS-DA model and two main components was calculated. The model parameters R2X=0.531, R2Y=0.388, Q2=0.0787. There are significantly differences between the two samples after intervention. There are11specific metabolites, including betaine, proline, leucine, glutamine, serine, histidine, alanine, nicotinamide nucleotide, isoleucine, aspartic acid, inositol. It is related to fatty acid metabolism, Krebs cycle, ornithine cycle, amino acid metabolism, pyruvate metabolism, gluconeogenesis, alanine cycle, inositol metabolism, steroid synthesis, purine pyrimidine metabolism, ammonia recovery, glycine and serine metabolism multiple metabolic pathway.
(3) The plasma1H NMR profiling of intervention group before and after intervention
We selected20volenteers (10in control group and10in intervention group) to study its metabolism characteristics. PCA method was used to build model and five main components was calculated. The model parameters R2X=0.725, Q2X=0.382, the model analyzed72.5%of the original data. Based on the data to establish the PLS-DA model and three main components was calculated. The model parameters R2X=0.502, R2Y=0.967, Q2X=0.721. There are significantly differences in intervention group before and after intervention. There are35specific metabolites, including renal steroids, coenzyme A, sitosterol, aldosterone, valine, cholesterol sulfate, ethanol, prostaglandin E1, isoleucine, alanine, gamma amino acid, glutamine, methionine, betaine, benzene alanine, phenylpropanolamine, alanine, glycerol acid, aspartic acid, serine, histidine, a-linolenic acid, prostaglandin D2,2-ethyl acrylate, different dimension A acid, sphingosine, urea, tyrosine, epinephrine, acetylamino, methyl hippuric acid, vanillic acid, hippuric acid, N-acetyl-tyrosine, p-hydroxybenzoic acid. It is related to fatty acid metabolism, the TCA cycle, ornithine cycle, catecholamine synthesis, amino acid metabolism, pyruvate metabolism, gluconeogenesis, synthesis of steroids, purine pyrimidine metabolism, metabolism of sphingolipids metabolism, retinol, ammonia recovery, synthetic phospholipids metabolism, glycine and serine metabolism, mitochondrial electron transport chain and multiple metabolic pathways.
Conclusion:Firstly, it indirectly shows that if the constitution can not be improved, it may convert to other constitutions.
Secondly, Bu zhong yi qi wan can significantly improve the qi-deficiency constitution.
Thirdly, the different metabolites of intervention group before and after intervention related to the citric acid cycle and bu zhong yi qi wan maybe regulate the disorder.
引文
[1]哈孝贤.漫谈中医的体质学说(三)[J].开卷有益：求医问药.2007(9)：28-29.
[2]王莉,何裕民.男女体质特点及其异同的研究——附2967例流调资料的模 糊识别分析[J].中国中医基础医学杂志.1998(2)：8-10.
[3]王琦,朱燕波.中国一般人群中医体质流行病学调查——基于全国9省市21948例流行病学调查数据[J].中华中医药杂志.2009,24(1)：7-12.
[4]中华中医药学会.中医体质分类与判定[J].世界中西医结合杂志.2009(4)：303-304.
[5]王琦.中医体质学[M].北京：中国医药科技出版社,1995.
[6]周敏,毛敏杰,朱敏,李凫坚,尹良胜,李锋.气虚体质及CYP2E1基因多态性与抗结核药物性肝损伤的关系[J].中西医结合肝病杂志.2012,22(5)：265-267.
[7]周小军,田道法,王士贞,阮岩,邱宝珊,张丽娟,卢标清.气虚体质之鼻咽癌高癌家系成员基因组稳定与修复基因表达[J].中医药信息.2009,26(5)：59-62.
[8]周俊亮,潘佩光,刘友章,潘奔前.蛋白质组学在气虚体质研究中的应用[J].世界中医药.2009,4(1)：46-48.
[9]李英帅,王琦,袁卓珺.阳虚体质者血清和尿液的核磁共振代谢组学[J].高等学校化学学报.2011,32(11)：2521-2527.
[10]郑军,温振英.健康儿童中医体质类型调查研究初探[J].中国医药学报.2000,15(2)：41-42.
[11]姚实林,吴芳斌,许霞,曹健,王文锐,李成国.1003例中医体质类型流行病学调查分析[J].安徽中医学院学报.2007,26(1)：10-13.
[12]雍小嘉,徐姗姗.青年气虚质与平和质人群声音特征对比分析[J].辽宁中医杂志.2013,40(6)：1121-1122.
[13]张晓,赵燕平,朱绘霖,刘颂豪.气虚质与平和质原穴生物电信号与主观疲劳量表的相关性研究[J].中医杂志.2013,54(19)：1649-1652.
[14]王睿林.气虚体质者外周血基因表达谱初步研究[D].硕士,北京中医药大学,2005.
[15]郝晓玲,吕秋军,周喆,苗爱东,王学军,苗明三,王升启.补中益气丸 等中成药的辐射防护作用[J].中华放射医学与防护杂志.2006,26(4)：366-369.
[16]涂小煌,陈志耀.补中益气丸联合易蒙停治疗大肠癌术后腹泻疗效观察[J].中国中西医结合杂志.2008,28(8)：738-741.
[17]沈前保.三磷酸腺菅、补中益气丸(汤)治疗脾虚气陷型胃粘膜脱垂30例临床分析[J].右江民族医学院学报.1999,21(4)：693.
[18]欧阳明子,谭为,刘艳艳,黄颖,肖雅,程静茹,.罗仁,赵晓山.补中益气丸对小鼠乳腺癌化疗相关性疲劳的影响[J].热带医学杂志.2013,13(5)：585-589.
[19]卿照前.补中益气丸对老年支气管哮喘缓解期患者肺功能的影响[J].湖南中医药大学学报.2007,27(3)：47-48.
[20]陈海蛟,唐志武,陈兴波.黄芪超细微粉与黄芪粗粉对雏鸡免疫功能的影响[J].安徽农业科学.2009,37(4)：1549-1550,1581.
[21]伍伟培.黄芪有效成分黄芪苷Ⅳ强心作用实验研究[J].中药材.2005,28(7)：591-592.
[22]阮长武,张代富,汪姗姗,王娟,方红贤.黄芪对缺血再灌注心肌细胞凋亡的影响[J].同济大学学报(医学版).2003,24(3)：182-184.
[23]李维祖,李卫平,尹艳艳,公惠玲,吴国翠,朱芬芳.黄芪总苷及黄芪甲苷对糖皮质激素诱导老前期大鼠记忆损伤的保护作用及机制[J].中国中药杂志.2009,34(2)：199-203.
[24]付荣国,周琳,聂萌,马力群,张星映,吕星,张红利.黄芪当归合剂对大鼠缺血性急性肾损伤的保护研究[J].中国中西医结合急救杂志.2006,13(1)：9-12.
[25]穆士卿,肖安平,李拴德,汪海波,李凯,吴云志.黄芪对急性脑创伤后局部脑皮质血流速度变化的影响[J].中国中西医结合急救杂志.2004,11(1)：45-46.
[26]裴林,刘唤荣,张少丹,丁春华,张建涛.黄芪对缺氧缺血性脑损伤幼鼠一氧化氮、丙二醛含量的影响[J].中国中西医结合急救杂志.2001,8(4)：222-224.
[27]王晓峰,李拴德,杨喜民,毛小林.应用黄芪治疗重型颅脑损伤的临床研究[J].中国中西医结合急救杂志.2001,8(1)：38-40.
[28]罗晓丽,杨侃,孙明.黄芪扶正汤对老年人免疫功能及氧化应激的影响[J].广东医学.2011,32(13)：1756-1758.
[29]余黎,曹宜,魏筱,许立,方泰惠.黄芪与红芪不同提取液免疫调节作用研究[J].安徽农业科学.2010,38(36)：20623-20624,20627.
[30]张振东,杨娟,吴兰芳,景永帅.神经营养因子样土党参多糖促进小鼠学习记忆作用的实验研究[J].时珍国医国药.2011,22(8)：1845-1847.
[31]张晓丹,刘琳,佟欣.党参、黄芪对中枢神经系统作用的比较研究[J].中草药.2003,34(9)：822-823.
[32]李贵荣,杨胜圆.党参多糖的提取及其对活性氧自由基的清除作用[J].化学世界.2001,42(8)：421-422,434.
[33]熊元君,李晓瑾,木拉提.新疆党参、潞党参对肠道影响[J].中药药理与临床.2000,16(2)：15.
[34]宋丹,王峥涛,李隆云,钟国跃.党参炔苷对胃溃疡模型大鼠胃黏膜损伤保护作用的研究[J].中国中医急症.2008,17(7)：963-964,986.
[35]杨光,李发胜,刘辉,咸丰.党参多糖对小鼠免疫功能的影响[J].中药药理与临床.2005,21(4)：39.
[36]张晓君,祝晨,胡黎,赖小平,莫建霞.党参多糖对小鼠免疫和造血功能的影响[J].中药新药与临床药理.2003,14(3)：174-176.
[37]柏长青,遆新宇,刘关键,李宁秀,董碧蓉.党参、黄芪提取物注射液对减轻肺癌患者化疗毒副反应和生存质量影响的研究[J].中国康复医学杂志.2006,21(8)：707-709.
[38]沈江龙.复方党参合用中药组方治疗眩晕症40例疗效观察[J].医药前沿.2011,1(18)：145-146.
[39]谭志鑫,肖本见,廖艳华.党参硒多糖对微血管反应性及抗氧化酶影响[J].中国公共卫生.2010,26(5)：557-558.
[40]王翕,刘玉瑛,史天良,张鸿翔,杨广文.白术挥发油抗实体瘤的作用研究[J].中国药物与临床.2002,2(4)：239-240.
[41]刘曼,叶峰,邱根全,章梅,王锐,何群英,蔡云.白术内酯Ⅰ对肿瘤恶病质患者细胞因子和肿瘤代谢因子的影响[J].第一军医大学学报.2005,25(10)：1308-1311.
[42]胡晓蕾,胡迎利,汪以真.白术及白术多糖对SD大鼠生长性能和免疫功能的影响[J].中国兽药杂志.2006,40(1)：2-6.
[43]徐丽珊,金晓玲,邵邻相.白术及白术多糖对小鼠学习记忆和抗氧化作用的影响[J].科技通报.2003,19(6)：513-515.
[44]李伟,文红梅,崔小兵,李璇,洪敏.白术健脾有效成分研究[J].南京中医药大学学报(自然科学版).2006,22(6)：366-367.
[45]朱金照.白术、霍香等中药对胃排空、肠推进影响的实验研究[J].中国中医基础医学杂志.2000,6(1)：21-22.
[46]刘兆平,杨陟华,朱茂祥,霍君生,卢承前,龚诒芬,陈君石.黑升麻的雌激素活性及其对人类乳腺癌MCF-7细胞雌激素受体水平的影响[J].卫生研究.2001,30(2)：77-80.
[47]李春梅,刘志峰,李敏,高永林,韩兵,刘珂.升麻提取物对去卵巢所致大鼠骨质疏松症的作用[J].中草药.2005,36(11)：1686-1688.
[48]欧立娟,刘启德.陈皮药理作用研究进展[J].中国药房.2006,17(10)：787-789.
[49]李伟,郑天珍,瞿颂义,田治峰,邱小青,丁光辉,卫玉玲.陈皮对小鼠胃排空及肠推进的影响[J].中药药理与临床.2002,18(2)：22-23.
[50]官福兰,王汝俊,王建华.陈皮及橙皮甙对小鼠胃排空、小肠推进功能的影响[J].中药药理与临床.2002,18(3)：7-9.
[51]李红芳,李丹明,瞿颂义,郑天珍,李伟,丁永辉,卫玉玲.枳实和陈皮对兔离体主动脉平滑肌条作用机理探讨[J].中成药.2001,23(9)：658-660.
[52]钱士辉,王佾先,亢寿海,杨念云,袁丽红.陈皮提取物体内抗肿瘤作用 及其对癌细胞增殖周期的影响[J].中国中药杂志.2003,28(12)：1167-1170.
[53]钱士辉,王佾先,亢寿海,杨念云,袁丽红.陈皮提取物体外抗肿瘤作用的研究[J].中药材.2003,26(10)：744-745.
[54]苏丹,鲁心安,秦德安.陈皮提取液抗衰老作用的实验研究[J].上海铁道大学学报.1999,20(9)：18-20.
[55]谢东浩,贾晓斌,蔡宝昌,张林松,袁冬平.北柴胡及春柴胡挥发油的解热作用比较[J].中国医院药学杂志.2007,27(4)：502-504.
[56]谢东浩,贾晓斌,蔡宝昌,张林松.北柴胡及春柴胡挥发油的抗炎镇痛作用的实验研究[J].药学与临床研究.2007,15(2)：108-110.
[57]李忻,靳耀英,张韫,杨鑫,李平,郭景珍.柴胡皂苷防治肝病机制研究进展[J].中国中西医结合杂志.2009,29(11)：1049-1051.
[58]张峰,曹仲伟,张学杰,李法曾.柴胡对大鼠慢性应激抑郁模型脑单胺类神经递质及其代谢物含量的影响[J].山东中医药大学学报.2005,29(3)：224-226.
[59]步世忠,许金廉,孙继虎,陈宜张,商权.柴胡皂甙d上调人急性早幼粒白血病细胞糖皮质激素受体mRNA对细胞生长的影响[J].中国中西医结合杂志.2000,20(5)：350-352.
[60]黄运生,谢炜,陈宝田.柴胡皂甙对癫痫大鼠脑电的影响[J].第一军医大学学报.2004,24(12)：1379-1381.
[61]朱兰香,刘世增,顾振纶.柴胡皂苷的药理作用及抗肝纤维化的应用[J].中草药.2002,33(10)：5-6.
[62]盖晓东,曾常茜,洪敏.柴胡逆转肝细胞癌多药耐药作用与相关机制的研究[J].高等学校化学学报.2005,26(8)：1446-1450.
[63]谢炜,鲍勇,于礼建.柴胡总皂甙及柴胡皂甙a、c、d对MES惊厥小鼠的影响[J].中药药理与临床.2006,22(1)：39-40.
[64]张建英,楚更五,刘秀萍,聂坚.当归散对正常和缩宫素处理大鼠离体子宫活动力的影响[J].中国实验方剂学杂志.2011,17(7)：201-203.
[65]宿树兰,华永庆,段金廒,王言才,鲍邢杰,唐于平.当归-川芎挥发性成 分与其抗子宫痉挛活性相关性分析[J].中国实验方剂学杂志.2009,15(2)：64-67.
[66]刘雪东,李伟东,蔡宝昌.当归化学成分及对心脑血管系统作用研究进展[J].南京中医药大学学报.2010,26(2)：155-157.
[67]李蕊蕊.生当归和酒当归抗氧化能力的比较[J].饲料博览.2011(8)：49-51.
[68]沈建芬,张又枝,肖军花,王嘉陵.当归A3活性部位对小鼠巨噬细胞环氧化酶-2活性及基因表达的影响[J].中国药理学通报.2011,27(11)：1574-1577.
[69]许琦,刘晓秋,王建华,唐惠琼,王汝俊.”脾虚”大鼠胃黏膜组织一氧化氮含量及补中益气汤的作用[J].中药药理与临床.2003,19(1)：7-8.
[70]Nabeshima S, Murata M, Hamada M, Chong Y, Yamaji K, Hayashi J. Maturation of monocyte-derived dendritic cells by Hochu-ekki-to, a traditional Japanese herbal medicine[J]. Int Immunopharmacol.2004,4(1):37-45.
[71]Tamura R, Takahashi H K, Xue D, Kubo S, Saito S, Nishibori M, Iwagaki H, Tanaka N. Enhanced effects of combined bu-zhong-yi-qi-tang (TJ-41) and interleukin-18 on the production of tumour necrosis factor-alpha and interferon-gamma in human peripheral blood mononuclear cells[J]. J Int Med Res. 2004,32(1):25-32.
[72]Kuroiwa A, Liou S, Yan H, Eshita A, Naitoh S, Nagayama A. Effect of a traditional Japanese herbal medicine, hochu-ekki-to (Bu-Zhong-Yi-Qi Tang), on immunity in elderly persons[J]. Int Immunopharmacol.2004,4(2):317-324.
[73]Kiyohara H, Nagai T, Munakata K, Nonaka K, Hanawa T, Kim S J, Yamada H. Stimulating effect of Japanese herbal (kampo) medicine, hochuekkito on upper respiratory mucosal immune system[J]. Evid Based Complement Alternat Med.2006, 3(4):459-467.
[74]李滨,齐凤琴,李燕敏,刘石磊.补中益气汤抗肿瘤作用的实验研究[J].中医药学报.2006,34(1)：22-23.
[75]Cho J M, Sato N, Kikuchi K. Prophylactic anti-tumor effect of Hochu-ekki-to (TJ41) by enhancing natural killer cell activity[J]. In Vivo.1991,5(4):389-391.
[76]俞传芳.补中益气汤加味治疗慢性萎缩性胃炎[J].上海中医药杂志.2004,38(8)：24-25.
[77]方文佳.补中益气汤合西药治疗肝硬化低蛋白血症62例[J].上海中医药杂志.2004,38(5)：33-34.
[78]乔根宝,许理忠,邬学群,叶洁.加味补中益气汤治疗颈性眩晕400例[J].上海中医药杂志.2004,38(7)：14-15.
[79]周仲瑛.补中益气汤临床新用[J].浙江中医药大学学报.2006,30(2)：156-157,162.
[80]王嘉梅.补中益气汤加味治疗药物流产后出血56例[J].四川中医.2004,22(12)：65.
[81]张美荣.健脾益气法治疗崩漏60例[J].陕西中医.2004,25(11)：970.
[82]Nicholson J K, Lindon J C, Holmes E.'Metabonomics':understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data[J]. Xenobiotica.1999, 29(11):1181-1189.
[83]Monton M R, Soga T. Metabolome analysis by capillary electrophoresis-mass spectrometry[J]. J Chromatogr A.2007,1168(1):237-246,236.
[84]邢世杰.乳腺癌血清代谢组学研究及特异性标志物筛选[D].硕士,哈尔滨工业大学,2012.
[85]Lv H, Palacios G, Hartil K, Kurland I J. Advantages of tandem LC-MS for the rapid assessment of tissue-specific metabolic complexity using a pentafluorophenylpropyl stationary phase[J]. J Proteome Res.2011,10(4): 2104-2112.
[86]赵维薇,许文涛,王龑,彭晓丽,黄昆仑.代谢组学研究技术及其应用[J].生物技术通报.2011(12)：57-64.
[87]Lu X, Zhao X, Bai C, Zhao C, Lu G, Xu G. LC-MS-based metabonomics analysis[J]. J Chromatogr B Analyt Technol Biomed Life Sci.2008,866(1):64-76.
[88]Boulesteix A L. PLS dimension reduction for classification with microarray data[J]. Stat Appl Genet Mol Biol.2004,3(1):33-36.
[89]许国旺,路鑫,杨胜利.代谢组学研究进展[J].中国医学科学院学报.2007,29(6)：701-711.
[90]王琦,朱燕波.中国一般人群中医体质流行病学调查——基于全国9省市21948例流行病学调查数据[J].中华中医药杂志.2009(01)：7-12.
[91]朱燕波,王琦,邓棋卫,蔡晶,宋晓鸿,闫雪.中医体质类型与高血压的相关性研究[J].中西医结合学报.2010,8(1)：40-45.
[92]闫镛,朱璞,张芳,顾娟娟,陈永强,苏惠娟.2型糖尿病患者中医体质类型与相关指标关系的分析[J].中医学报.2010,25(6)：1154-1156.
[93]张超群,白介辰.从流行病学探讨瘦人多阴虚[J].湖北中医杂志.1992,14(6)：27-28.
[94]朱燕波,王琦,吴承玉,庞国明,赵健雄,沈世林,夏仲元,闫雪.18805例中国成年人中医体质类型与超重和肥胖关系的Logistic回归分析[J].中西医结合学报.2010,8(11)：1023-1028.
[95]张振贤,张烨,吴丽丽,陈敏,周长林,沈辉.慢性疲劳综合征与中医体质及社会心理危险因素相关性调查[J].辽宁中医杂志.2011,38(1)：22-26.
[96]毕建璐,陈晶,于冰琰,邓卫,赖名慧,罗仁,林家省,赵晓山.亚健康大学生阴虚及阳虚体质特征分析[J].中国公共卫生.2011,27(1)：87-89.
[97]陈晶,魏敏,戴红芳,赵晓山,李玉萍,李纪强,罗仁.亚健康大学生中医体质现状调查[J].中国公共卫生.2011,27(11)：1488-1489.
[98]张杰能,罗仁.中医体质与2型糖尿病之初探[J].内蒙古中医药.2010,29(4)：128-129.
[99]郑良琴,毕建璐,陈晶,于冰琰,邓卫,赖名慧,罗仁,林家省,赵晓山.女大学生亚健康状态与气郁质关系的调查分析[J].热带医学杂志.2011,11(5)：593-595.
[100]郑良琴,毕建璐,占春旺,邓卫,林家省,罗仁,赵晓山.大学女生中医体质与原发性痛经的相关性研究[J].中医药导报.2011,17(1)：27-29.
[101]陈晶.亚健康自评量表的编制与大学生亚健康中医体质研究[D].博士,南方医科大学,2009.
[102]邱玉明.疲劳型亚健康的中医体质特征及其尿液代谢组学研究[D].博士, 南方医科大学,2011.
[103]欧阳涛,宋剑南,林谦,李林,苗阳,牛晓红,金红,陈冰.冠心病体质表型和低密度脂蛋白受体基因Avall位点多态性关系的研究[J].中国中医基础医学杂志.2005,11(7)：521-523.
[104]沈乃莹,金宇安,韩静,闫征,何培英,王申五HLA-DQA1等位基因与急性脑梗塞多种中医体质类型的相关性研究[J].中医杂志.2001,42(4)：237-238.
[105]Wang Q, Yao S. Molecular basis for cold-intolerant yang-deficient constitution of traditional Chinese medicine[J]. Am J Chin Med.2008,36(5):827-834.
[106]王琦,姚实林,董静,吴宏东,吴承玉,夏仲元,石鹤峰,庞国明,邓祺卫,赵健雄,蔡晶,崔正植.阳虚体质者内分泌及免疫功能变化[J].中西医结合学报.2008,6(12)：1226-1232.
[107]魏敏.肾阴虚证和肾阳虚证基因表达谱的比较研究[D].博士,南方医科大学,2009.
[108]毕建璐.肾阳虚证和肾阴虚证血浆蛋白表达谱的比较研究[D].硕士,南方医科大学,2011.
[109]薛琪,聂晓莉,罗仁,赵晓山,杨晓云,毛秋平.基于OPLS-DA的高脂血症血浆1H-NMR研究[J].实用医学杂志.2010,26(21)：3874-3877.
[1]朱嵘.《亚健康中医临床指南》解读[J].中国中医药现代远程教育.2009,7(2)：5-6.
[2]中华中医药学会.中医体质分类与判定[J].世界中西医结合杂志.2009,4(4)：303-304.
[3]陈晶,于冰琰,魏敏,戴红芳,李纪强,赵晓山,罗仁.第2代亚健康调查表信度与效度评价[J].中国公共卫生.2009,25(1)：37-38.
[4]Pender N J, Walker S N, Sechrist K R, Frank-Stromborg M. Predicting health-promoting lifestyles in the workplace [J]. Nurs Res.1990,39(6):326-332.
[5]Teng H L, Yen M, Fetzer S. Health promotion lifestyle profile-Ⅱ:Chinese version short form[J]. J Adv Nurs.2010,66(8):1864-1873.
[6]陈良英,余丽娟.中年知识分子健康促进生活方式及影响因素的研究[J].中华现代护理杂志.2009,15(34)：3605-3607.
[7]崔仁善,李春玉.延边地区中年女性健康促进生活方式影响因素分析[J].延边大学医学学报.2009,32(4)：264-267.
[8]王琦,王睿林,李英帅.中医体质学学科发展述评[J].中华中医药杂志.2007,22(9)：627-630.
[9]王琦,朱燕波.中国一般人群中医体质流行病学调查——基于全国9省市21948例流行病学调查数据[J].中华中医药杂志.2009,24(1)：7-12.
[10]周敏,毛敏杰,朱敏,李凫坚,尹良胜,李锋.气虚体质及CYP2E1基因多态性与抗结核药物性肝损伤的关系[J].中西医结合肝病杂志.2012,22(5)：265-267.
[11]周小军,田道法,王士贞,阮岩,邱宝珊,张丽娟,卢标清.气虚体质之鼻咽癌高癌家系成员基因组稳定与修复基因表达[J].中医药信息.2009,26(5)：59-62.
[12]周俊亮,潘佩光,刘友章,潘奔前.蛋白质组学在气虚体质研究中的应用[J].世界中医药.2009,4(1)：46-48.
[13]李英帅,王琦,袁卓珺.阳虚体质者血清和尿液的核磁共振代谢组学[J].高等学校化学学报.2011,32(11)：2521-2527.
[14]毕建璐,陈晶,于冰琰,邓卫,赖名慧,罗仁,林家省,赵晓山.亚健康大学生阴虚及阳虚体质特征分析[J].中国公共卫生.2011,27(1)：87-89.
[15]陈晶,魏敏,戴红芳,赵晓山,李玉萍,李纪强,罗仁.亚健康大学生中医体质现状调查[J].中国公共卫生.2011,27(11)：1488-1489.
[16]张杰能,罗仁.中医体质与2型糖尿病之初探[J].内蒙古中医药.2010,29(4)： 128-129.
[17]郑良琴,毕建璐,陈晶,于冰琰,邓卫,赖名慧,罗仁,林家省,赵晓山.女大学生亚健康状态与气郁质关系的调查分析[J].热带医学杂志.2011,11(5)：593-595.
[18]郑良琴,毕建璐,占春旺,邓卫,林家省,罗仁,赵晓山.大学女生中医体质与原发性痛经的相关性研究[J].中医药导报.2011,17(1)：27-29.
[19]陈晶.亚健康自评量表的编制与大学生亚健康中医体质研究[D].博士,南方医科大学,2009.
[20]邱玉明.疲劳型亚健康的中医体质特征及其尿液代谢组学研究[D].博士,南方医科大学,2011.
[21]郝晓玲,吕秋军,周喆,苗爱东,王学军,苗明三,王升启.补中益气丸等中成药的辐射防护作用[J].中华放射医学与防护杂志.2006,26(4)：366-369.
[22]涂小煌,陈志耀.补中益气丸联合易蒙停治疗大肠癌术后腹泻疗效观察[J].中国中西医结合杂志.2008,28(8)：738-741.
[23]沈前保.三磷酸腺苷、补中益气丸(汤)治疗脾虚气陷型胃粘膜脱垂30例临床分析[J].右江民族医学院学报.1999,21(4)：693.
[24]欧阳明子,谭为,刘艳艳,黄颖,肖雅,程静茹,罗仁,赵晓山.补中益气丸对小鼠乳腺癌化疗相关性疲劳的影响[J].热带医学杂志.2013,13(5)：585-589.
[25]卿照前.补中益气丸对老年支气管哮喘缓解期患者肺功能的影响[J].湖南中医药大学学报.2007,27(3)：47-48.
[1]张涛,董宽虎.蛋氨酸、胆碱和甜菜碱在甲基供体中的相互关系[J].中国家禽.2003,25(12)：39-41.
[2]王云霞,李文君,王润霞.甜菜碱、胆碱和蛋氨酸的协同作用[J].黑龙江畜牧兽医.2003(11)：54-55.
[3]卢建军,邹晓庭.甜菜碱对蛋鸡血清卵黄前体物水平的影响[J].浙江大学学报(农业与生命科学版).2006,32(3)：287-291.
[4]Brouns F, Beckers E, Wagenmakers A J, Saris W H. Ammonia accumulation during highly intensive long-lasting cycling:individual observations [J]. Int J Sports Med.1990, 11(Suppl 2):S78-S84.
[5]Broberg S, Sahlin K. Adenine nucleotide degradation in human skeletal muscle during prolonged exercise[J]. J Appl Physiol.1989,67(1):116-122.
[6]Lewis D A, Volk D W, Hashimoto T. Selective alterations in prefrontal cortical GABA neurotransmission in schizophrenia:a novel target for the treatment of working memory dysfunction[J]. Psychopharmacology (Berl).2004,174(1):143-150.
[7]Castellano C, Cestari V, Cabib S, Puglisi-Allegra S. Strain-dependent effects of post-training GABA receptor agonists and antagonists on memory storage in mice[J]. Psychopharmacology (Berl).1993,111(2):134-138.
[8]江波.GABA((?)-氨基丁酸)——一种新型的功能食品因子[J].中国食品学报.2008,8(2)：1-4.
[9]Guezennec C Y, Abdelmalki A, Serrurier B, Merino D, Bigard X, Berthelot M, Pierard C, Peres M. Effects of prolonged exercise on brain ammonia and amino acids[J]. Int J Sports Med.1998,19(5):323-327.
[10]Hashimoto K, Fukushima T, Shimizu E, Okada S, Komatsu N, Okamura N, Koike K, Koizumi H, Kumakiri C, Imai K, Iyo M. Possible role of D-serine in the pathophysiology of Alzheimer's disease[J]. Prog Neuropsychopharmacol Biol Psychiatry.2004,28(2):385-388.
[11]Andersen J D, Pouzet B. Spatial memory deficits induced by perinatal treatment of rats with PCP and reversal effect of D-serine [J]. Neuropsychopharmacology.2004, 29(6):1080-1090.
[12]Andersen J M, Fonnum F, Myhrer T. D-Serine alleviates retrograde amnesia of a visual discrimination task in rats with a lesion of the perirhinal cortex[J]. Brain Res. 2003,979(1):240-244.
[2]王莉,何裕民.男女体质特点及其异同的研究——附2967例流调资料的模 糊识别分析[J].中国中医基础医学杂志.1998(2)：8-10.
[3]王琦,朱燕波.中国一般人群中医体质流行病学调查——基于全国9省市21948例流行病学调查数据[J].中华中医药杂志.2009,24(1)：7-12.
[4]中华中医药学会.中医体质分类与判定[J].世界中西医结合杂志.2009(4)：303-304.
[5]王琦.中医体质学[M].北京：中国医药科技出版社,1995.
[6]周敏,毛敏杰,朱敏,李凫坚,尹良胜,李锋.气虚体质及CYP2E1基因多态性与抗结核药物性肝损伤的关系[J].中西医结合肝病杂志.2012,22(5)：265-267.
[7]周小军,田道法,王士贞,阮岩,邱宝珊,张丽娟,卢标清.气虚体质之鼻咽癌高癌家系成员基因组稳定与修复基因表达[J].中医药信息.2009,26(5)：59-62.
[8]周俊亮,潘佩光,刘友章,潘奔前.蛋白质组学在气虚体质研究中的应用[J].世界中医药.2009,4(1)：46-48.
[9]李英帅,王琦,袁卓珺.阳虚体质者血清和尿液的核磁共振代谢组学[J].高等学校化学学报.2011,32(11)：2521-2527.
[10]郑军,温振英.健康儿童中医体质类型调查研究初探[J].中国医药学报.2000,15(2)：41-42.
[11]姚实林,吴芳斌,许霞,曹健,王文锐,李成国.1003例中医体质类型流行病学调查分析[J].安徽中医学院学报.2007,26(1)：10-13.
[12]雍小嘉,徐姗姗.青年气虚质与平和质人群声音特征对比分析[J].辽宁中医杂志.2013,40(6)：1121-1122.
[13]张晓,赵燕平,朱绘霖,刘颂豪.气虚质与平和质原穴生物电信号与主观疲劳量表的相关性研究[J].中医杂志.2013,54(19)：1649-1652.
[14]王睿林.气虚体质者外周血基因表达谱初步研究[D].硕士,北京中医药大学,2005.
[15]郝晓玲,吕秋军,周喆,苗爱东,王学军,苗明三,王升启.补中益气丸 等中成药的辐射防护作用[J].中华放射医学与防护杂志.2006,26(4)：366-369.
[16]涂小煌,陈志耀.补中益气丸联合易蒙停治疗大肠癌术后腹泻疗效观察[J].中国中西医结合杂志.2008,28(8)：738-741.
[17]沈前保.三磷酸腺菅、补中益气丸(汤)治疗脾虚气陷型胃粘膜脱垂30例临床分析[J].右江民族医学院学报.1999,21(4)：693.
[18]欧阳明子,谭为,刘艳艳,黄颖,肖雅,程静茹,.罗仁,赵晓山.补中益气丸对小鼠乳腺癌化疗相关性疲劳的影响[J].热带医学杂志.2013,13(5)：585-589.
[19]卿照前.补中益气丸对老年支气管哮喘缓解期患者肺功能的影响[J].湖南中医药大学学报.2007,27(3)：47-48.
[20]陈海蛟,唐志武,陈兴波.黄芪超细微粉与黄芪粗粉对雏鸡免疫功能的影响[J].安徽农业科学.2009,37(4)：1549-1550,1581.
[21]伍伟培.黄芪有效成分黄芪苷Ⅳ强心作用实验研究[J].中药材.2005,28(7)：591-592.
[22]阮长武,张代富,汪姗姗,王娟,方红贤.黄芪对缺血再灌注心肌细胞凋亡的影响[J].同济大学学报(医学版).2003,24(3)：182-184.
[23]李维祖,李卫平,尹艳艳,公惠玲,吴国翠,朱芬芳.黄芪总苷及黄芪甲苷对糖皮质激素诱导老前期大鼠记忆损伤的保护作用及机制[J].中国中药杂志.2009,34(2)：199-203.
[24]付荣国,周琳,聂萌,马力群,张星映,吕星,张红利.黄芪当归合剂对大鼠缺血性急性肾损伤的保护研究[J].中国中西医结合急救杂志.2006,13(1)：9-12.
[25]穆士卿,肖安平,李拴德,汪海波,李凯,吴云志.黄芪对急性脑创伤后局部脑皮质血流速度变化的影响[J].中国中西医结合急救杂志.2004,11(1)：45-46.
[26]裴林,刘唤荣,张少丹,丁春华,张建涛.黄芪对缺氧缺血性脑损伤幼鼠一氧化氮、丙二醛含量的影响[J].中国中西医结合急救杂志.2001,8(4)：222-224.
[27]王晓峰,李拴德,杨喜民,毛小林.应用黄芪治疗重型颅脑损伤的临床研究[J].中国中西医结合急救杂志.2001,8(1)：38-40.
[28]罗晓丽,杨侃,孙明.黄芪扶正汤对老年人免疫功能及氧化应激的影响[J].广东医学.2011,32(13)：1756-1758.
[29]余黎,曹宜,魏筱,许立,方泰惠.黄芪与红芪不同提取液免疫调节作用研究[J].安徽农业科学.2010,38(36)：20623-20624,20627.
[30]张振东,杨娟,吴兰芳,景永帅.神经营养因子样土党参多糖促进小鼠学习记忆作用的实验研究[J].时珍国医国药.2011,22(8)：1845-1847.
[31]张晓丹,刘琳,佟欣.党参、黄芪对中枢神经系统作用的比较研究[J].中草药.2003,34(9)：822-823.
[32]李贵荣,杨胜圆.党参多糖的提取及其对活性氧自由基的清除作用[J].化学世界.2001,42(8)：421-422,434.
[33]熊元君,李晓瑾,木拉提.新疆党参、潞党参对肠道影响[J].中药药理与临床.2000,16(2)：15.
[34]宋丹,王峥涛,李隆云,钟国跃.党参炔苷对胃溃疡模型大鼠胃黏膜损伤保护作用的研究[J].中国中医急症.2008,17(7)：963-964,986.
[35]杨光,李发胜,刘辉,咸丰.党参多糖对小鼠免疫功能的影响[J].中药药理与临床.2005,21(4)：39.
[36]张晓君,祝晨,胡黎,赖小平,莫建霞.党参多糖对小鼠免疫和造血功能的影响[J].中药新药与临床药理.2003,14(3)：174-176.
[37]柏长青,遆新宇,刘关键,李宁秀,董碧蓉.党参、黄芪提取物注射液对减轻肺癌患者化疗毒副反应和生存质量影响的研究[J].中国康复医学杂志.2006,21(8)：707-709.
[38]沈江龙.复方党参合用中药组方治疗眩晕症40例疗效观察[J].医药前沿.2011,1(18)：145-146.
[39]谭志鑫,肖本见,廖艳华.党参硒多糖对微血管反应性及抗氧化酶影响[J].中国公共卫生.2010,26(5)：557-558.
[40]王翕,刘玉瑛,史天良,张鸿翔,杨广文.白术挥发油抗实体瘤的作用研究[J].中国药物与临床.2002,2(4)：239-240.
[41]刘曼,叶峰,邱根全,章梅,王锐,何群英,蔡云.白术内酯Ⅰ对肿瘤恶病质患者细胞因子和肿瘤代谢因子的影响[J].第一军医大学学报.2005,25(10)：1308-1311.
[42]胡晓蕾,胡迎利,汪以真.白术及白术多糖对SD大鼠生长性能和免疫功能的影响[J].中国兽药杂志.2006,40(1)：2-6.
[43]徐丽珊,金晓玲,邵邻相.白术及白术多糖对小鼠学习记忆和抗氧化作用的影响[J].科技通报.2003,19(6)：513-515.
[44]李伟,文红梅,崔小兵,李璇,洪敏.白术健脾有效成分研究[J].南京中医药大学学报(自然科学版).2006,22(6)：366-367.
[45]朱金照.白术、霍香等中药对胃排空、肠推进影响的实验研究[J].中国中医基础医学杂志.2000,6(1)：21-22.
[46]刘兆平,杨陟华,朱茂祥,霍君生,卢承前,龚诒芬,陈君石.黑升麻的雌激素活性及其对人类乳腺癌MCF-7细胞雌激素受体水平的影响[J].卫生研究.2001,30(2)：77-80.
[47]李春梅,刘志峰,李敏,高永林,韩兵,刘珂.升麻提取物对去卵巢所致大鼠骨质疏松症的作用[J].中草药.2005,36(11)：1686-1688.
[48]欧立娟,刘启德.陈皮药理作用研究进展[J].中国药房.2006,17(10)：787-789.
[49]李伟,郑天珍,瞿颂义,田治峰,邱小青,丁光辉,卫玉玲.陈皮对小鼠胃排空及肠推进的影响[J].中药药理与临床.2002,18(2)：22-23.
[50]官福兰,王汝俊,王建华.陈皮及橙皮甙对小鼠胃排空、小肠推进功能的影响[J].中药药理与临床.2002,18(3)：7-9.
[51]李红芳,李丹明,瞿颂义,郑天珍,李伟,丁永辉,卫玉玲.枳实和陈皮对兔离体主动脉平滑肌条作用机理探讨[J].中成药.2001,23(9)：658-660.
[52]钱士辉,王佾先,亢寿海,杨念云,袁丽红.陈皮提取物体内抗肿瘤作用 及其对癌细胞增殖周期的影响[J].中国中药杂志.2003,28(12)：1167-1170.
[53]钱士辉,王佾先,亢寿海,杨念云,袁丽红.陈皮提取物体外抗肿瘤作用的研究[J].中药材.2003,26(10)：744-745.
[54]苏丹,鲁心安,秦德安.陈皮提取液抗衰老作用的实验研究[J].上海铁道大学学报.1999,20(9)：18-20.
[55]谢东浩,贾晓斌,蔡宝昌,张林松,袁冬平.北柴胡及春柴胡挥发油的解热作用比较[J].中国医院药学杂志.2007,27(4)：502-504.
[56]谢东浩,贾晓斌,蔡宝昌,张林松.北柴胡及春柴胡挥发油的抗炎镇痛作用的实验研究[J].药学与临床研究.2007,15(2)：108-110.
[57]李忻,靳耀英,张韫,杨鑫,李平,郭景珍.柴胡皂苷防治肝病机制研究进展[J].中国中西医结合杂志.2009,29(11)：1049-1051.
[58]张峰,曹仲伟,张学杰,李法曾.柴胡对大鼠慢性应激抑郁模型脑单胺类神经递质及其代谢物含量的影响[J].山东中医药大学学报.2005,29(3)：224-226.
[59]步世忠,许金廉,孙继虎,陈宜张,商权.柴胡皂甙d上调人急性早幼粒白血病细胞糖皮质激素受体mRNA对细胞生长的影响[J].中国中西医结合杂志.2000,20(5)：350-352.
[60]黄运生,谢炜,陈宝田.柴胡皂甙对癫痫大鼠脑电的影响[J].第一军医大学学报.2004,24(12)：1379-1381.
[61]朱兰香,刘世增,顾振纶.柴胡皂苷的药理作用及抗肝纤维化的应用[J].中草药.2002,33(10)：5-6.
[62]盖晓东,曾常茜,洪敏.柴胡逆转肝细胞癌多药耐药作用与相关机制的研究[J].高等学校化学学报.2005,26(8)：1446-1450.
[63]谢炜,鲍勇,于礼建.柴胡总皂甙及柴胡皂甙a、c、d对MES惊厥小鼠的影响[J].中药药理与临床.2006,22(1)：39-40.
[64]张建英,楚更五,刘秀萍,聂坚.当归散对正常和缩宫素处理大鼠离体子宫活动力的影响[J].中国实验方剂学杂志.2011,17(7)：201-203.
[65]宿树兰,华永庆,段金廒,王言才,鲍邢杰,唐于平.当归-川芎挥发性成 分与其抗子宫痉挛活性相关性分析[J].中国实验方剂学杂志.2009,15(2)：64-67.
[66]刘雪东,李伟东,蔡宝昌.当归化学成分及对心脑血管系统作用研究进展[J].南京中医药大学学报.2010,26(2)：155-157.
[67]李蕊蕊.生当归和酒当归抗氧化能力的比较[J].饲料博览.2011(8)：49-51.
[68]沈建芬,张又枝,肖军花,王嘉陵.当归A3活性部位对小鼠巨噬细胞环氧化酶-2活性及基因表达的影响[J].中国药理学通报.2011,27(11)：1574-1577.
[69]许琦,刘晓秋,王建华,唐惠琼,王汝俊.”脾虚”大鼠胃黏膜组织一氧化氮含量及补中益气汤的作用[J].中药药理与临床.2003,19(1)：7-8.
[70]Nabeshima S, Murata M, Hamada M, Chong Y, Yamaji K, Hayashi J. Maturation of monocyte-derived dendritic cells by Hochu-ekki-to, a traditional Japanese herbal medicine[J]. Int Immunopharmacol.2004,4(1):37-45.
[71]Tamura R, Takahashi H K, Xue D, Kubo S, Saito S, Nishibori M, Iwagaki H, Tanaka N. Enhanced effects of combined bu-zhong-yi-qi-tang (TJ-41) and interleukin-18 on the production of tumour necrosis factor-alpha and interferon-gamma in human peripheral blood mononuclear cells[J]. J Int Med Res. 2004,32(1):25-32.
[72]Kuroiwa A, Liou S, Yan H, Eshita A, Naitoh S, Nagayama A. Effect of a traditional Japanese herbal medicine, hochu-ekki-to (Bu-Zhong-Yi-Qi Tang), on immunity in elderly persons[J]. Int Immunopharmacol.2004,4(2):317-324.
[73]Kiyohara H, Nagai T, Munakata K, Nonaka K, Hanawa T, Kim S J, Yamada H. Stimulating effect of Japanese herbal (kampo) medicine, hochuekkito on upper respiratory mucosal immune system[J]. Evid Based Complement Alternat Med.2006, 3(4):459-467.
[74]李滨,齐凤琴,李燕敏,刘石磊.补中益气汤抗肿瘤作用的实验研究[J].中医药学报.2006,34(1)：22-23.
[75]Cho J M, Sato N, Kikuchi K. Prophylactic anti-tumor effect of Hochu-ekki-to (TJ41) by enhancing natural killer cell activity[J]. In Vivo.1991,5(4):389-391.
[76]俞传芳.补中益气汤加味治疗慢性萎缩性胃炎[J].上海中医药杂志.2004,38(8)：24-25.
[77]方文佳.补中益气汤合西药治疗肝硬化低蛋白血症62例[J].上海中医药杂志.2004,38(5)：33-34.
[78]乔根宝,许理忠,邬学群,叶洁.加味补中益气汤治疗颈性眩晕400例[J].上海中医药杂志.2004,38(7)：14-15.
[79]周仲瑛.补中益气汤临床新用[J].浙江中医药大学学报.2006,30(2)：156-157,162.
[80]王嘉梅.补中益气汤加味治疗药物流产后出血56例[J].四川中医.2004,22(12)：65.
[81]张美荣.健脾益气法治疗崩漏60例[J].陕西中医.2004,25(11)：970.
[82]Nicholson J K, Lindon J C, Holmes E.'Metabonomics':understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data[J]. Xenobiotica.1999, 29(11):1181-1189.
[83]Monton M R, Soga T. Metabolome analysis by capillary electrophoresis-mass spectrometry[J]. J Chromatogr A.2007,1168(1):237-246,236.
[84]邢世杰.乳腺癌血清代谢组学研究及特异性标志物筛选[D].硕士,哈尔滨工业大学,2012.
[85]Lv H, Palacios G, Hartil K, Kurland I J. Advantages of tandem LC-MS for the rapid assessment of tissue-specific metabolic complexity using a pentafluorophenylpropyl stationary phase[J]. J Proteome Res.2011,10(4): 2104-2112.
[86]赵维薇,许文涛,王龑,彭晓丽,黄昆仑.代谢组学研究技术及其应用[J].生物技术通报.2011(12)：57-64.
[87]Lu X, Zhao X, Bai C, Zhao C, Lu G, Xu G. LC-MS-based metabonomics analysis[J]. J Chromatogr B Analyt Technol Biomed Life Sci.2008,866(1):64-76.
[88]Boulesteix A L. PLS dimension reduction for classification with microarray data[J]. Stat Appl Genet Mol Biol.2004,3(1):33-36.
[89]许国旺,路鑫,杨胜利.代谢组学研究进展[J].中国医学科学院学报.2007,29(6)：701-711.
[90]王琦,朱燕波.中国一般人群中医体质流行病学调查——基于全国9省市21948例流行病学调查数据[J].中华中医药杂志.2009(01)：7-12.
[91]朱燕波,王琦,邓棋卫,蔡晶,宋晓鸿,闫雪.中医体质类型与高血压的相关性研究[J].中西医结合学报.2010,8(1)：40-45.
[92]闫镛,朱璞,张芳,顾娟娟,陈永强,苏惠娟.2型糖尿病患者中医体质类型与相关指标关系的分析[J].中医学报.2010,25(6)：1154-1156.
[93]张超群,白介辰.从流行病学探讨瘦人多阴虚[J].湖北中医杂志.1992,14(6)：27-28.
[94]朱燕波,王琦,吴承玉,庞国明,赵健雄,沈世林,夏仲元,闫雪.18805例中国成年人中医体质类型与超重和肥胖关系的Logistic回归分析[J].中西医结合学报.2010,8(11)：1023-1028.
[95]张振贤,张烨,吴丽丽,陈敏,周长林,沈辉.慢性疲劳综合征与中医体质及社会心理危险因素相关性调查[J].辽宁中医杂志.2011,38(1)：22-26.
[96]毕建璐,陈晶,于冰琰,邓卫,赖名慧,罗仁,林家省,赵晓山.亚健康大学生阴虚及阳虚体质特征分析[J].中国公共卫生.2011,27(1)：87-89.
[97]陈晶,魏敏,戴红芳,赵晓山,李玉萍,李纪强,罗仁.亚健康大学生中医体质现状调查[J].中国公共卫生.2011,27(11)：1488-1489.
[98]张杰能,罗仁.中医体质与2型糖尿病之初探[J].内蒙古中医药.2010,29(4)：128-129.
[99]郑良琴,毕建璐,陈晶,于冰琰,邓卫,赖名慧,罗仁,林家省,赵晓山.女大学生亚健康状态与气郁质关系的调查分析[J].热带医学杂志.2011,11(5)：593-595.
[100]郑良琴,毕建璐,占春旺,邓卫,林家省,罗仁,赵晓山.大学女生中医体质与原发性痛经的相关性研究[J].中医药导报.2011,17(1)：27-29.
[101]陈晶.亚健康自评量表的编制与大学生亚健康中医体质研究[D].博士,南方医科大学,2009.
[102]邱玉明.疲劳型亚健康的中医体质特征及其尿液代谢组学研究[D].博士, 南方医科大学,2011.
[103]欧阳涛,宋剑南,林谦,李林,苗阳,牛晓红,金红,陈冰.冠心病体质表型和低密度脂蛋白受体基因Avall位点多态性关系的研究[J].中国中医基础医学杂志.2005,11(7)：521-523.
[104]沈乃莹,金宇安,韩静,闫征,何培英,王申五HLA-DQA1等位基因与急性脑梗塞多种中医体质类型的相关性研究[J].中医杂志.2001,42(4)：237-238.
[105]Wang Q, Yao S. Molecular basis for cold-intolerant yang-deficient constitution of traditional Chinese medicine[J]. Am J Chin Med.2008,36(5):827-834.
[106]王琦,姚实林,董静,吴宏东,吴承玉,夏仲元,石鹤峰,庞国明,邓祺卫,赵健雄,蔡晶,崔正植.阳虚体质者内分泌及免疫功能变化[J].中西医结合学报.2008,6(12)：1226-1232.
[107]魏敏.肾阴虚证和肾阳虚证基因表达谱的比较研究[D].博士,南方医科大学,2009.
[108]毕建璐.肾阳虚证和肾阴虚证血浆蛋白表达谱的比较研究[D].硕士,南方医科大学,2011.
[109]薛琪,聂晓莉,罗仁,赵晓山,杨晓云,毛秋平.基于OPLS-DA的高脂血症血浆1H-NMR研究[J].实用医学杂志.2010,26(21)：3874-3877.
[1]朱嵘.《亚健康中医临床指南》解读[J].中国中医药现代远程教育.2009,7(2)：5-6.
[2]中华中医药学会.中医体质分类与判定[J].世界中西医结合杂志.2009,4(4)：303-304.
[3]陈晶,于冰琰,魏敏,戴红芳,李纪强,赵晓山,罗仁.第2代亚健康调查表信度与效度评价[J].中国公共卫生.2009,25(1)：37-38.
[4]Pender N J, Walker S N, Sechrist K R, Frank-Stromborg M. Predicting health-promoting lifestyles in the workplace [J]. Nurs Res.1990,39(6):326-332.
[5]Teng H L, Yen M, Fetzer S. Health promotion lifestyle profile-Ⅱ:Chinese version short form[J]. J Adv Nurs.2010,66(8):1864-1873.
[6]陈良英,余丽娟.中年知识分子健康促进生活方式及影响因素的研究[J].中华现代护理杂志.2009,15(34)：3605-3607.
[7]崔仁善,李春玉.延边地区中年女性健康促进生活方式影响因素分析[J].延边大学医学学报.2009,32(4)：264-267.
[8]王琦,王睿林,李英帅.中医体质学学科发展述评[J].中华中医药杂志.2007,22(9)：627-630.
[9]王琦,朱燕波.中国一般人群中医体质流行病学调查——基于全国9省市21948例流行病学调查数据[J].中华中医药杂志.2009,24(1)：7-12.
[10]周敏,毛敏杰,朱敏,李凫坚,尹良胜,李锋.气虚体质及CYP2E1基因多态性与抗结核药物性肝损伤的关系[J].中西医结合肝病杂志.2012,22(5)：265-267.
[11]周小军,田道法,王士贞,阮岩,邱宝珊,张丽娟,卢标清.气虚体质之鼻咽癌高癌家系成员基因组稳定与修复基因表达[J].中医药信息.2009,26(5)：59-62.
[12]周俊亮,潘佩光,刘友章,潘奔前.蛋白质组学在气虚体质研究中的应用[J].世界中医药.2009,4(1)：46-48.
[13]李英帅,王琦,袁卓珺.阳虚体质者血清和尿液的核磁共振代谢组学[J].高等学校化学学报.2011,32(11)：2521-2527.
[14]毕建璐,陈晶,于冰琰,邓卫,赖名慧,罗仁,林家省,赵晓山.亚健康大学生阴虚及阳虚体质特征分析[J].中国公共卫生.2011,27(1)：87-89.
[15]陈晶,魏敏,戴红芳,赵晓山,李玉萍,李纪强,罗仁.亚健康大学生中医体质现状调查[J].中国公共卫生.2011,27(11)：1488-1489.
[16]张杰能,罗仁.中医体质与2型糖尿病之初探[J].内蒙古中医药.2010,29(4)： 128-129.
[17]郑良琴,毕建璐,陈晶,于冰琰,邓卫,赖名慧,罗仁,林家省,赵晓山.女大学生亚健康状态与气郁质关系的调查分析[J].热带医学杂志.2011,11(5)：593-595.
[18]郑良琴,毕建璐,占春旺,邓卫,林家省,罗仁,赵晓山.大学女生中医体质与原发性痛经的相关性研究[J].中医药导报.2011,17(1)：27-29.
[19]陈晶.亚健康自评量表的编制与大学生亚健康中医体质研究[D].博士,南方医科大学,2009.
[20]邱玉明.疲劳型亚健康的中医体质特征及其尿液代谢组学研究[D].博士,南方医科大学,2011.
[21]郝晓玲,吕秋军,周喆,苗爱东,王学军,苗明三,王升启.补中益气丸等中成药的辐射防护作用[J].中华放射医学与防护杂志.2006,26(4)：366-369.
[22]涂小煌,陈志耀.补中益气丸联合易蒙停治疗大肠癌术后腹泻疗效观察[J].中国中西医结合杂志.2008,28(8)：738-741.
[23]沈前保.三磷酸腺苷、补中益气丸(汤)治疗脾虚气陷型胃粘膜脱垂30例临床分析[J].右江民族医学院学报.1999,21(4)：693.
[24]欧阳明子,谭为,刘艳艳,黄颖,肖雅,程静茹,罗仁,赵晓山.补中益气丸对小鼠乳腺癌化疗相关性疲劳的影响[J].热带医学杂志.2013,13(5)：585-589.
[25]卿照前.补中益气丸对老年支气管哮喘缓解期患者肺功能的影响[J].湖南中医药大学学报.2007,27(3)：47-48.
[1]张涛,董宽虎.蛋氨酸、胆碱和甜菜碱在甲基供体中的相互关系[J].中国家禽.2003,25(12)：39-41.
[2]王云霞,李文君,王润霞.甜菜碱、胆碱和蛋氨酸的协同作用[J].黑龙江畜牧兽医.2003(11)：54-55.
[3]卢建军,邹晓庭.甜菜碱对蛋鸡血清卵黄前体物水平的影响[J].浙江大学学报(农业与生命科学版).2006,32(3)：287-291.
[4]Brouns F, Beckers E, Wagenmakers A J, Saris W H. Ammonia accumulation during highly intensive long-lasting cycling:individual observations [J]. Int J Sports Med.1990, 11(Suppl 2):S78-S84.
[5]Broberg S, Sahlin K. Adenine nucleotide degradation in human skeletal muscle during prolonged exercise[J]. J Appl Physiol.1989,67(1):116-122.
[6]Lewis D A, Volk D W, Hashimoto T. Selective alterations in prefrontal cortical GABA neurotransmission in schizophrenia:a novel target for the treatment of working memory dysfunction[J]. Psychopharmacology (Berl).2004,174(1):143-150.
[7]Castellano C, Cestari V, Cabib S, Puglisi-Allegra S. Strain-dependent effects of post-training GABA receptor agonists and antagonists on memory storage in mice[J]. Psychopharmacology (Berl).1993,111(2):134-138.
[8]江波.GABA((?)-氨基丁酸)——一种新型的功能食品因子[J].中国食品学报.2008,8(2)：1-4.
[9]Guezennec C Y, Abdelmalki A, Serrurier B, Merino D, Bigard X, Berthelot M, Pierard C, Peres M. Effects of prolonged exercise on brain ammonia and amino acids[J]. Int J Sports Med.1998,19(5):323-327.
[10]Hashimoto K, Fukushima T, Shimizu E, Okada S, Komatsu N, Okamura N, Koike K, Koizumi H, Kumakiri C, Imai K, Iyo M. Possible role of D-serine in the pathophysiology of Alzheimer's disease[J]. Prog Neuropsychopharmacol Biol Psychiatry.2004,28(2):385-388.
[11]Andersen J D, Pouzet B. Spatial memory deficits induced by perinatal treatment of rats with PCP and reversal effect of D-serine [J]. Neuropsychopharmacology.2004, 29(6):1080-1090.
[12]Andersen J M, Fonnum F, Myhrer T. D-Serine alleviates retrograde amnesia of a visual discrimination task in rats with a lesion of the perirhinal cortex[J]. Brain Res. 2003,979(1):240-244.
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