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基于NEI、PTS的复杂性疾病共性机制与“异病同证、同证同理”性的研究
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摘要
目的:
     复杂性疾病,是指由多个基因位点共同参与,且和环境因素相互作用决定表型的遗传病,如肿瘤、糖尿病、哮喘等,此已成为危害人类健康的重要因素,是现代生物学的研究热点之一。随着现代生命科学研究的迅速发展,医学界对人体生理机能整体调节方式的探索倍受关注。神经内分泌免疫网络的研究是现代医学前沿领域之一,随着对神经系统-内分泌系统和免疫系统之间相互作用、相互依赖复杂关系的深入研究,已有大量实验证实神经系统、内分泌系统、免疫系统的组织细胞均可表达神经递质、激素、细胞因子及其相应受体,并以此作为生物学物质基础,相互间进行双向性的信息传递,通过相互刺激、相互制约、正负反馈,达到系统内部的自我调制和相对稳定,在细胞、分子和基因水平上构成一个动态平衡的调节网络。研究发现复杂性疾病虽然在诊断和治疗等方面各异,但都拥有现代医学病理生理的共同机理,如血栓前状态改变等。
     作为我国传统医学之一的维吾尔医学在复杂性疾病的认识和治疗方面拥有独特的理论、组方和疗效。维医学历来非常重视人体本身的统一性、完整性及其与自然界的相互关系,认为人体是一个有机整体,构成人体的各个脏器、组织或器官在结构上不可分割,在功能上相互协调,在病理上相互影响。从维医病理学角度来看,复杂性疾病的发生有共同的基础,常由异常黑胆质所致。维吾尔医在临床实践中,对复杂性疾病的治疗,经辨证分型属异常异常黑胆质证者,首先使用异常黑胆质成熟剂对异常黑胆质体液进行调理和堆积,然后使用异常黑胆质清除剂,使已成熟好的异常黑胆质体液排出体外,从而使气质复原,体液平衡,为本脏的治疗奠定基础,最后针对具体的病因进行相应的治疗,取得了良好治疗效果。
     复杂性疾病之所以复杂,原因就在于其超越了一个部位、一个器官或一个系统的范畴,而影响到多个系统以至整个机体的平衡状态。由此,在研究疾病本身的基础上,有必要研究疾病背后整体的生物学基础紊乱状态,并加以干预以改善之,使疾病的发展处于一种孤立状态,从而改善疾病的恶化,延缓疾病的进程。为此,有必要找出复杂性病种背后的共性机制及其共同证型加以研究。
     本研究对恶性肿瘤、2型糖尿病、哮喘等复杂性疾病患者血清/或血浆中有关神经-内分泌-免疫学指标及血栓前状态分子标志物进行检测,试从神经-内分泌-免疫网络、血栓前状态等方面,阐明复杂性疾病的共性机制及系统疾病性。并以维吾尔医理论为指导,按照体液分型标准,对所有患者进行维医异常体液分型,寻找其中最常见的维医异常体液证型,探讨该证型与神经-内分泌-免疫网络及血栓前状态的关系,试图找出复杂性疾病背后的共性机制,阐述复杂性疾病的“异病同证”“同证同理”性。
     方法:
     (1)复杂性疾病患者维吾尔医分型:收集2006年1月至2009年8月在新疆医科大学各附属医院诊治的恶性肿瘤患者287例、2型糖尿病患者267例、哮喘发作期患者307例,共861例患者,经体格检查正常的健康志愿者33例。观察患者的舌象、脉象、纳寐、二便、肌肤、口味、眼部变化并询问患者主观症状进行维吾尔医辨证分型。
     (2)神经内分泌免疫网络相关指标测定:对632例患者(218例恶性肿瘤患者、174例2型糖尿病患者、240例哮喘患者)及33例健康志愿者进行以下指标的测定:用流式细胞仪对外周血CD4~+、CD8~+、CD4~+/CD8~+及NK进行检测;采用放射免疫分析法对血清白细胞介素-1(IL-1)、白细胞介素-6(IL-6)、肿瘤坏死因子α(TNF-α)、促肾上腺皮质激素(ACTH)及皮质醇(CORT)进行检测;用液相色谱法检测血浆去甲肾上腺素(NE)和多巴胺(DA)含量。
     (3)血栓前状态分子标志物的测定:对229例患者(69恶性肿瘤患者、93例2型糖尿病患者、67例哮喘患者)及33例健康志愿者进行以下指标的测定:用流式细胞术测定患者血小板表面CD41、CD62p;用放射免疫法检测血清ET-1;用ELISA方法检测血浆t-PA、PAI-1;全自动血凝分析仪检测凝血四项即血浆纤维蛋白原(FIB)水平、活化部分凝血活酶时间(APTT)、凝血酶原时间(PT)、凝血酶时间(TT)。
     (4)统计方法:计量资料以均数±标准差(x±s)表示,如果符合正态性、方差齐性,多组间比较选用完全随机设计多组间单因素方差分析;如果不符合正态性、方差齐性,则采用秩和检验;使用SPSS13.0统计软件进行统计学分析,检验水准α=0.05。P<0.05时说明差异有统计学意义。
     结果:
     (1)复杂性疾病组与健康对照组神经内分泌免疫网络相关指标(T淋巴细胞亚群、NK细胞水平,IL-1β、IL-6、TNF-α、ACTH、CORT、DA及NE含量)的测定结果显示:与正常对照组相比,肿瘤、2-型糖尿病、哮喘患者组CD4~+细胞数及CD4~+/CD8~+比值下降(P<0.05),CD8~+细胞数升高(P>0.05),NK细胞数下降(P<0.05);IL-1β、IL-6含量升高(P<0.05),TNF-α含量在不同复杂性疾病中高低有别,在2-型糖尿病组、哮喘组升高(P<0.05),恶性肿瘤组下降(P<0.05);ACTH含量下降,在2-型糖尿病组与哮喘组中有统计学意义(P<0.05),恶性肿瘤组无统计学意义(P>0.05);CORT含量下降(P<0.05),NE、DA含量升高(P<0.05)。上述指标在各疾病组间比较无差异(P>0.05)。
     (2)复杂性疾病组与健康对照组血栓前状态分子标志物(CD41、CD62P、t-PA、PAI-1及ET-1)测定结果显示:与正常对照组相比,肿瘤、2-型糖尿病、哮喘患者组血小板表面CD41的表达无统计学意义(P>0.05),血小板表面CD62p的表达及血浆PAI-1水平升高(P<0.05);t-PA的含量降低(P<0.05);ET-1水平升高,在肿瘤组中有统计学意义(P<0.05),在2-型糖尿病组与哮喘组中无统计学意义(P>0.05);APTT时间缩短(P<0.05);FIB浓度升高(P<0.05);PT时间缩短,在肿瘤组、2-型糖尿病有统计学意义(P<0.05),哮喘组无统计学意义(P>0.05);TT时间无差异(P>0.05)。上述指标在各疾病组间比较无差异(P>0.05)。
     (3)在复杂性疾病861例患者中,属异常黑胆质证的有476例,非异常黑胆质证为385例;异常黑胆质证占全部病例的55.28%,非异常黑胆质证占44.72%,异常黑胆质证所占比例大于非异常黑胆质证。其中,糖尿病患者中,异常黑胆质证141例,占糖尿病患者总数的52.18%,肿瘤患者中,异常黑胆质证181例,占肿瘤患者总数的63.05%,哮喘患者中,异常黑胆质证183例,占哮喘患者总数的59.58%。
     (4)复杂性疾病维吾尔医各证型组与健康对照组神经内分泌免疫网络相关指标的测定结果显示:与正常对照组相比,CD4~+、NK细胞水平在异常黑胆质证患者组和非异常黑胆质证患者组下降(P<0.05),与非异常黑胆质证组相比,异常黑胆质证组CD4~+及NK细胞水平下降明显(P<0.05);与正常对照组相比,IL-1β、IL-6、TNF-α含量在异常黑胆质证患者组和非异常黑胆质证患者组升高(P<0.05),与非异常黑胆质证组相比,异常黑胆质证组IL-1β、IL-6含量升高明显(P<0.05);与正常对照组相比, ACTH、CORT含量在异常黑胆质证患者组和非异常黑胆质证患者组下降(P<0.05),DA及NE含量在异常黑胆质证患者组和非异常黑胆质证患者组升高(P<0.05),与非异常黑胆质证组相比,异常黑胆质证组NE含量升高明显(P<0.05)。
     (5)复杂性疾病维吾尔医各证型组与健康对照组血栓前状态分子标志物的测定结果显示:与正常对照组相比,ET、CD62p、PAI-1水平在异常黑胆质证组和非异常黑胆质证组升高(P<0.05),与非异常黑胆质证组相比,异常黑胆质证组ET、CD62p升高明显(P<0.05);t-PA水平在异常黑胆质证组和非异常黑胆质证组下降(P<0.05);CD41水平在各组间比较无差异(P>0.05);与正常对照组相比,FIB水平在异常黑胆质证组和非异常黑胆质证组升高(P<0.05),与非异常黑胆质证组相比,异常黑胆质证组FIB水平升高明显(P<0.05); PT、APTT时间在异常黑胆质证组和非异常黑胆质证组缩短(P<0.05),TT在各组间比较无差异(P>0.05)。
     结论:
     (1)研究发现,复杂性疾病具有神经-内分泌-免疫网络紊乱、血栓前状态改变的共性。神经-内分泌-免疫网络紊乱、血栓前状态改变可能是复杂性疾病共存的部分病生基础。
     (2)复杂性疾病神经-内分泌-免疫网络紊乱、血栓前状态改变的整个过程的连续性及关联性提示复杂性疾病不是单纯的局部性病变,而是机体全身性的变化,复杂性疾病具有系统疾病性。
     (3)研究发现,复杂性疾病具有“异病同证”“同证同理”性。异常黑胆质证是复杂性疾病常见的维医证型,在复杂性疾病异常胆液质证、异常粘液质证、异常血液质证、异常黑胆质证中,异常黑胆质证型占55.28%。与复杂性疾病非异常黑胆质证(异常胆液质证、异常粘液质证、异常血液质证)患者相比,异常黑胆质证患者的神经-内分泌-免疫网络紊乱、血栓前状态改变程度更为严重,表现在免疫功能更加紊乱(CD4~+、NK细胞水平下降,IL-1β、IL-6含量升高,P<0.05),交感神经兴奋更加明显(NE含量升高,P<0.05),血管内皮细胞损伤更加严重(ET水平升高,P<0.05)、血小板活化性增强(CD62p水平升高,P<0.05)、血液粘度增高(FIB水平升高,P<0.05)。
     (4)研究发现,异常黑胆质的产生过程可能包含着神经-内分泌-免疫网络功能的紊乱;包含着机体神经系统、内分泌系统、免疫系统之间相互制约、相互协调机制的失衡;包含着血管内皮细胞的损伤、血小板的活化、血液粘度的增高、纤溶功能的降低。发现上述变化与维医体内异常体液堆积、“燃烧”最终产生异常黑胆质的过程相似或相同。此方面的研究正在进行中。
Objective:
     Complex Diseases such as malignant cancer, asthma and diabetes, caused by thecommon interaction of multiple genes and environmental factors, are threatening humanbeings, and becoming hotspot in modern biology.
     In recent years scientists put forward a new way of studying the human body as onewhole system. The essence of the neuro-endocrine-immune network (NEI) is to keep thehomeostasis and balance of organism by the three interdependent systems--nervous,endocrine and immune systems. The interaction between them is mediated by the nervoustransmitters, hormones and cytokines. It has been discovered that cancer, diabetes andasthma have many differences on their diagnosis’ and treatments, however all of thesehave pathophysiological change, prethrombotic state (PTS) in common.
     Uyghur medicines always stress the importance of considering the human body as awhole unit. It has its unique theoriy and treatment methods for complex diseases. Thefluid theory in Uighur traditional medicine describes the following: body fluid is the basicmatter during the vital life and physiological activity; it is produced and utilizedcontinuously and provides the energy for the vital activity. The abnormal savda as thefinal pathological production of abnormal sapra、abnormal kan, abnormal blgam is relatedclosely to the complex diseases.
     Complex diseases are complex, the reason lies in its beyond a part, an organs or asystem, and also affect the balance of whole systems.Thus, It is necessary to study theoverall biological disorder behind the basis of disease and to improve it, make thedevelopment of the disease in an isolated state, so as to improve the deterioration of thedisease, and slow down the progress of the disease. Therefore, it is necessary to find outthe common mechanism and common type on complex diseases.
     In this study we tested the expression of CD41, CD62p on platelets, the level ofplasma tissue plasminogen activator (t-PA) and its inhibiter (PAI-1), the level of endothelin (ET-1), Activated partial thromboplastin time (APTT), Fibrinogen (FIB),Prothrombin time (PT), Thrombin time (TT), the level of CD4~+, CD8~+, CD4~+/CD8~+,Narural killer (NK), Interleukin-1(IL-1β), Interleukin-6(IL-6), Tumor necrosis factor-α(TNF-α), Adrenocorticotropic-hormone (ACTH), Corticosterone (CORT), Dopamine(DA) and Norepinephrine (NE) on patients of malignant cancer, cardiovascular diseasesand diabetes try to find out common mechanism and systematicness of complex diseasesfrom neuro-endocrine-immune network and prethrombotic state. we classified all thepatients according to body fluids standard in Uyghur Medicine tried to seek out one of themost common and most serious abnormal type then discussed the relationship betweenthat type and the neuro-endocrine-immune network、prethrombotic state, by using thismethod we tried to discover the common mechanism and common type in UyghurMedicine for complex diseases.
     Materials and Methods
     (1) in Uyghur Medicine of complex diseases287cases of malignant cancer,267casesof diabetes,307cases of asthma were provided by the Affiliated Hospital of XinjiangMedical University. Patients were classified according to Chinese Traditional and UygurMedicine fluid theory. For the control group,33cases of healthy individuals werechecked to make sure they did not have hypertension, diabetes or other heart, liver, kidneyand blood diseases. All the patients were classified according to body fluids standard inUyghur Medicine from the change of tongue, pulse condition, diet, sleep, defecate,urinate, skin, taste, eyes.
     (2) Determination on the neuroendocrine immune network related indicators:632patients (218patients of malignant tumor,174cases of diabetes,240cases of asthma) and33healthy volunteers were detected the following index: Flow cytometer technology wasutilized to detect the serum level count of CD3~+、CD4~+and CD8~+, and NK cell.Radio-immune technology was utilized to detect the serum level of cytokines includingIL-1β (Interleukin-1β)、IL-6、TNF-α (Tumor necrosis factor-α), and hormones likeACTH (adrenocorticotrophic hormone). Liquid Chromatography was utilized to detectthe plazma level of DA (dopamine) and NE (norepinephrine).
     (3) Determination on the prethrombotic state marks:228patients (69cases ofmalignant cancer,93cases of diabetes,67cases of asthma) and33healthy volunteerswere detected the following index: The expression of CD41, CD62p on platelets, the levelof plasma tissue plasminogen activator (t-PA) and its inhibiter (PAI-1), the level of endothelin (ET-1), Activated partial thromboplastin time (APTT), Fibrinogen (FIB),Prothrombin time (PT) and Thrombin time (TT) were tested by using Flow Cytometer,ELISA method, radioimmunoassay method and auto coagulometer.
     (4) Statistical analysis: SPSS13.0was used in the study. The data were expressed asmeans±standard deviation. Significance levels of a comparison were determined by usingANOVA. The difference between means were considered to be statistically significant ifP<0.05.
     Results
     (1) The neuroendocrine immune network related indicators (between complexdiseases and control group) Compared to the control group, the amount of CD4~+,CD4~+/CD8~+, NK in complex diseases group were decreased (P<0.05); the amount ofCD8~+was increased; The level of IL-1β、IL-6in complex diseases group were increased(P<0.05); the amount of TNF-α were different in complex diseases, it was increased indiabetes and asthma group and was decreased in malignant tumor group; The level ofACTH and CORTin complex diseases group were decreased (P<0.05), The level of NE、DA in complex diseases group were increased (P<0.05), there was no difference onabove indexes between each complex diseases.
     (2) The prethrombotic state marks (between complex diseases and control group)Compared to the control group, the average amount of CD62P, the level of plasma PAI-1were increased (P<0.05) in complex diseases group, The level of t-PA was decreased incomplex diseases group (P<0.05); the level of ET-1were increased in the tumor group(P<0.05), there was no significant change in the level of ET-1between control group andcomplex diseases group. APTT were significantly shorter in complex diseases group (P<0.05), the level of plasma FIB was increased in complex diseases group (P<0.05).PTwere significantly shorter in malignant cancer, diabetes and hypertension group, therewas no chang onTT, there was no difference on above indexes between each complexdiseases.
     (3) There were476cases of Abnormal Savda patients and385cases of nonAbnormal Savda patients among861cases of complex disease patients; among the totalclinical cases, the ratio of Abnormal Savda patients was55.28%, the ratio of nonAbnormal Savda patients was44.72%, the ratio of Abnormal Savda patients is muchbigger than the ratio of non Abnormal Savda patients.
     (4) The neuroendocrine immune network related indicators (between Abnormal Savda group of complex diseases and control group) Compared to control group, theaverage expression of CD4~+、NK were significantly increased both in Abnormal Savdagroup and non Abnormal Savda group (P<0.05); Compared to non Abnormal Savdagroup, the average expression of CD4~+、NK were significantly increased in AbnormalSavda group (P<0.05); Compared to control group, the level of IL-1β、IL-6、TNF-α weresignificantly increased both in Abnormal Savda group and non Abnormal Savda group (P<0.05); Compared to non Abnormal Savda group, the the leve of IL-1β、IL-6weresignificantly increased in Abnormal Savda group (P<0.05); Compared to control group,the the leve of ACTH、CORT were significantly decreased both in Abnormal Savda groupand non Abnormal Savda group (P<0.05); Compared to control group, the the leve ofDA、NE were significantly increased both in Abnormal Savda group and non AbnormalSavda group (P<0.05); Compared to non Abnormal Savda group, the leve of NE wassignificantly increased in Abnormal Savda group (P<0.05);
     (5) The prethrombotic state marks (between Abnormal Savda group of complexdiseases and control group) Compared to the control group, the average amount of CD62P,the plasma PAI-1and serum ET-1were significantly increased both in Abnormal Savdagroup and non Abnormal Savda group (P<0.05). Compared to non Abnormal Savdagroup, the average expression of ET、CD62p were significantly increased in AbnormalSavda group (P<0.05); Compared to the control group, the level of t-PA wassignificantly increased both in Abnormal Savda group and non Abnormal Savda group (P<0.05). There was no difference in the amount of CD41between the Abnormal Savdagroup and the control group (P>0.05). Compared to the control group, the level of FIBwas significantly increased both in Abnormal Savda group and non Abnormal Savdagroup (P<0.05), Compared to non Abnormal Savda group, the level of FIB wassignificantly increased in Abnormal Savda group (P<0.05), Compared to the controlgroup, the APTT and PT were significantly shorter both in the Abnormal Savda group andnon Abnormal Savda group (P<0.05); there was no significant change in TT between thetwo groups (P>0.05).
     Conclusion
     (1) Although the External symptoms of complex diseases are different, there arecommonness in pathogenesis on adjustment of neuroendocrine immune network, theCellular immune were reduced, the endocrine hormone levels were suppressed,Sympathetic activation, There is prethromboitic state both in complex diseases. Neuroendocrine-immune network disorder and Prethromboitic state might be one of thePathophysiological basis of complex diseases.
     (2) The continuity and relevance of the whole process prompts that complex diseaseis not simply localized lesions, but the body's systemic change, That is to say complexdisease is not the only change in a certain organ but the overall change of the body.
     (3) Abnormal Savda Syndrome is the major type of complex diseases.Neuroendocrine-immune network disorder are more obvious on Abnormal SavdaSyndrome of complex diseases. Reflected in the disorderness of the immune function andobviousness of Sympathetic nerve activity. Prethromboitic state exists in the AbnornalSavda syndrom of complex diseases, Manifested in the injury of Vascular endothelial cell,activation of platelet, increasness of blood viscosity and reduction of fibrinolytic function.
     (4) During the Abnormal Savda generation process may be contained thedisfunctionness of the nervous system, endocrine system, immune system. Brokenbalance between nervous system, endocrine system, immune system might be one of themechanisms of the pathology of abnormal Savda. Immune dysfunction may be animportant feature of abnormal Savda Syndrome. Prethromboitic state exists in theAbnornal Savda syndrom of complex diseases, Manifested in the injury of Vascularendothelial cell, activation of platelet, increasness of blood viscosity and reduction offibrinolytic function.Vascular endothelial cell injury, platelet activation, increased bloodviscosity and reduced fibrinolytic function are the reasons for the formation of blood clot.This process also might be the one for the accumulation and burning of Abnormal Savdawhich eventually leads to the Abormal Savda syndrome.
引文
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