摘要
孙树椿筋伤学术经验总结
孙树椿首席研究员是国内著名骨伤科专家,全国老中医药专家学术经验指导老师,国家骨伤科重点专科学科带头人、国家骨伤科学学术带头人。近四十余年来,孙老一直从事骨伤科的临床、科研和教学工作,善于灵活运用中西医两种方法,进行诊断和治疗骨伤科筋伤疾病;积极提倡中医手法的治疗,努力挖掘和发扬祖国传统医学特色。现将孙老对于筋伤疾病的相关学术思想概述如下。
1孙氏筋伤学科思想
孙氏筋伤手法主要源于清代宫廷上驷院,并广泛吸收现代医学的成就,结合自身经验加以提炼,而形成的独具特色的孙氏筋伤学科思想。即:理论与临床并重;辨病与辨证结合;内治与外治相辅;动静结合,主动为主;筋伤辨治,气血为要;倡导规范,发挥优势。
孙老常教育学生“心到、眼到,不如手到”,只有多接触病人,才能找到手下的感觉,才能把理论运用在临床诊治中;孙老也强调,病人是我们最好的老师,加强跟病人的交流能更好地提高手法的技艺,这些都是书本上所没有的。孙老自始至终把理论与临床并重的理念贯穿其中,身体力行的做好榜样的带头作用。
孙老认为,有病就有证,辨证才能识病,两者是密不可分的。临床诊治时,既要辨病,又要辨证;只有病、证合参,才能选用适当方药、恰当的手法。西医辨病与中医辨证相结合只是临床诊治中的一个模式,我们应该立足中医辨证论治的同时,合理利用现代医学的理论、检测手段,拓宽延长自己的诊断视野,在中医理论指导下,去分析观察疾病内在的病因、病机和演变规律。
孙老强调治疗要兼顾局部与整体,审证求因。由于筋伤疾病多以局部损伤为主,因此治疗时,既要注意局部,又要兼顾全身,然后选取恰当的手法,以舒筋活络,解痉止痛,这样才能兼顾局部与整体。
孙老指出,动和静是对立统一的,静是为了更好的动,动也是为了更好的静,两者之间体现了辨证关系。静的作用,是使伤病肢体得到休养,有利于损伤组织的修复和肢体功能活动的还原。
孙老指出,筋伤辨治应以气血辩证为主。他认为,气血循行全身,内伤上下、皮肉筋骨、五脏六腑、四肢百骸,无所不在,故人体损伤无论何处,首当其冲伤及气血。
中医手法应该有一套成熟的、规范的、标准的操作准则。要制定严格的标准的操作规范,孙老号召中医骨伤界同仁共同努力。虽然困难重重,但这代表着学科的发展方向,中医事业的发展方向。
2孙氏筋伤辨证体系
孙老以中医望,闻,问,切四诊为基础,结合自己多年的临床经验,提炼出一套适合于筋伤疾病的独具特色的辩证体系。
问诊在诊病过程中是必不可少的,它在四诊中占有重要地位。自古至今各代医家对问诊都非常重视。通过问诊可以了解患者的自觉症状,疾病损伤的时间、体位,损伤时患者的情况,治疗的经过,以及既往的健康状况,家族史等。
望诊的内容包括观察人体的神、色、形、态。孙老强调,筋伤望诊还应重视对局部损伤区的观察。
闻诊除了听病人的语言、呼吸、喘息、咳嗽、呕吐、呃逆等一般内容以外,主要是听患者受伤部位发出的声响。
孙氏筋伤诊治主要以摸诊为主。摸诊为切诊的一种方式。摸诊与脉诊不同,摸诊是通过医生双手触摸、按压损伤的局部或全身的一种检查方法,可提供重要的诊断依据。摸诊可以对损伤的部位的情况有个较为明确的了解,尤其在缺少X线设备的情况下,更具有重要意义。
3孙氏筋伤学术特色
孙老师从著名老中医刘寿山老先生,尽得刘老先生理伤正骨手法真传。孙老在继承刘老先生经验的同时,运用现代的解剖生理学和病理生理学知识对其进行了规范整理。在保持疗效的基础上,简化精练了手法,使之便于学习掌握和推广应用,逐渐形成具有孙氏特点的筋伤手法。孙老通过四十余年的临床经验总结,博采众长,形成具有孙氏特色的筋伤治疗方法,其中包括筋伤手法(尤其是颈椎手法)、专病专方及功能练功。
孙老认为,手法的疗效靠的是手法本身,正如《医宗金鉴·正骨心法要旨》所言:“诚以手本血肉之体,其宛转运用之妙,可以一己之卷舒,高下徐疾,轻重开合,能达病者之血气凝滞,皮肉肿痛,筋骨挛折,与情之苦欲也”,手法治疗必须要做到“一旦临证,知其体相,识其部位”。孙老指出“筋喜柔不喜刚”,“法之所施,使患者不知其苦,方称为手法也”,在手法运用上尤其强调轻柔绵软、外柔内刚,力量由轻渐重,治疗中使患者在并不感到痛苦的情况下即获得症状的缓解或痊愈。孙氏手法的独到之处正是在于轻、巧、柔、和。
颈性心绞痛的相关实验研究
实验一颈性心肌缺血动物模型的建立
1材料和方法
30只家兔随机分为3组,每组10只,正常组、模型组和假手术组。采用切断颈5~6、颈6~7、颈7~胸1棘上韧带、剥离椎旁肌、轴向旋转错位颈6~7椎体的方法建立颈性心肌缺血动物模型,然后在造模后4、8、12周时拍摄颈椎正侧位X光片,观察不同摄片时期动物的颈椎椎间隙、颈椎曲度、椎间孔缩小程度、椎体前后缘骨质增生以及颈椎上下关节突骨质增生等的变化情况,并进行X线评分分析;通过在造模前、造模后1、2、4、6、8、10、12周分别检测实验动物的心电图改变情况,观察是否因颈椎失稳而出现心电图改变。ECG改变主要观察各波波形、P-R间期、Q-T间期、ST段偏移、QRS波群、T波的形状方向等方面的变化情况。
2实验结果及分析
2.1各组家兔颈椎X线定量观察结果及分析:模型组颈椎X线改变的积分值在实验4周时已经明显高于正常组和假手术组,差异具有显著性(P<0.01),而且这种差异随着时间的延长更加显著,在第12周时最为明显(P<0.01),而正常组和假手术组的颈椎X线改变积分之间比较无显著性差异(P>0.05)。说明模型组通过手术造成颈椎失稳后,颈椎发生了类似颈椎病的明显退变,而且这种退变失稳的情况随着时间延长,逐渐加重。
2.2各组家兔心电图结果及分析:颈椎错位前后及全部实验周期内,三组家兔心电图P波时间测量结果均比较接近,相互间比较无显著性差异(P>0.05),说明家兔颈椎错位失稳后,对心房传导的电激动过程无明显影响。模型组颈椎错位后的P-R间期较错位前明显延长,相互间比较具有显著性差异(P<0.05);模型组的P-R间期与正常组和假手术组相当于错位后及全部实验周期内时测得的P-R问期相比较也具有显著性差异(P<0.05),且时间均延长,说明颈椎错位后影响了心脏房室传导的电激动过程,使心房开始兴奋到心室兴奋所需的时间延长。模型组错位后的QRS时间较错位前明显延长,差异具有显著性(P<0.05);在实验8周以前,模型组心电图QRS时间与正常组和假手术组相当于错位后测得的QRS时间相比较明显延长,具有显著性差异(P<0.05),8周后差异更为显著(P<0.01),说明颈椎错位失稳影响了心室兴奋传导的电激动过程。模型组颈椎错位失稳后心电图Q-T间期较错位前明显延长,差异具有显著性(P<0.05);模型组错位后的Q-T间期与正常组和假手术组相当于错位后测得的Q-T间期相比较也明显延长,具有显著性差异(P<0.05);在总的实验周期内,正常组和假手术组的Q-T间期与模型组相比较均具有显著性差异(P<0.05),说明颈椎错位失稳影响了心室开始除极至完成全部复极的电激动过程。模型组在手术使颈椎失稳后,全部动物立即都出现了心电图的ST段下移,有的表现为T波低平或倒置,与正常组和假手术组比较具有极显著性差异(P<0.001)。随着颈椎错位时间的延长,模型组家兔出现的心电图ST—T改变一直持续,而正常组和假手术组在整个实验过程中均未出现ST-T异常,说明通过颈椎手术,在造成椎体错位失稳的情况即引起了家兔心肌缺血,颈性心绞痛动物模型造模成功。
实验二颈性心肌缺血家兔血浆神经肽类物质变化的观察
1材料和方法
建立颈椎错位失稳心肌缺血动物模型,实验结束时3组动物静脉取血进行神经肽类物质NPY、ET、CGRP、ANP含量的放射免疫检测。
2实验结果及分析
模型组家兔的血浆NPY、ET、ANP明显高于正常组和假手术组,有显著性差异(P<0.05,P<0.01);正常组和假手术组的血浆NPY、ET、ANP之间无显著性差异(P>0.05);模型组家兔的血浆CGRP明显低于正常组和假手术组,有显著性差异(P<0.05,P<0.01);正常组和假手术组的血浆CGRP之间无显著性差异(P>0.05)。说明颈椎失稳刺激了动物的交感神经纤维,使交感神经活性增高,同时也使与去甲肾上腺素共存于交感神经系统的NPY合成和释放增加,NPY的直接缩血管作用和其增强其它缩血管活性物质如去甲肾上腺素等的作用,以及能抑制腺苷等内源性扩血管物质对冠脉的舒张效应的作用可使冠状动脉痉挛而致心肌缺血;交感神经兴奋释放大量的儿茶酚胺,可同时直接促进血管内皮合成和分泌ET,也可使椎基底动脉供血不足,使脑缺血、缺氧而刺激脑血管内皮细胞及神经胶质细胞ET-mRNA表达,使ET分泌增加,另外颈椎长期错位失稳局部骨赘增生可导致椎动脉受压,经壁压力增高,内皮细胞产生大量的ET,ET强大的缩血管功能使冠状动脉痉挛而致心肌缺血;血浆NPY释放增加,加强了对心脏感觉神经的调节,引起CGRP的合成和释放减少,CGRP在NPY突触前调节的作用下,合成及释放的减少必然会使心脏正常的变时和变力以及扩张冠状动脉的功能受到影响,使心脏的功能受到影响,心肌供血不足;交感神经兴奋后儿茶酚胺的释放,可导致肺、心肌缺血缺氧以及肺小血管及全身皮肤毛细血管收缩,回心血量增多,右心房压增高,心钠素释放增多,心钠素的这种升高可能是机体的一种保护性代偿反应。这也反映了心源性激素和局部激素的分泌,对维持内环境的相对稳定的反馈机制。
实验三颈性心肌缺血家兔颈椎间盘炎症反应情况的观察
1材料和方法
造模后12周取C6、7颈椎间盘,进行HE染色和免疫组化染色,观察颈性心肌缺血家兔椎间盘周围组织的病理表现。
2实验结果及分析
模型组共取得10个间盘。中央部位均为髓核组织及纤维软骨组织,经HE染色显示,部分标本软骨有增生、灶状变性,均未见血管组织及炎症细胞;边缘部位为透明或纤维软骨组织,经HE染色显示,存在灶状增生、粘液变性。其中,7个(70%)间盘周围组织有炎症反应,表现为小血管增生、炎症细胞浸润、组织水肿及纤维素样渗出等。3个(30%)间盘周围组织未见血管增生及炎细胞浸润等炎症表现。假手术组与正常组各取得10个间盘,经HE染色显示为软骨组织,其边缘均未见血管增生及炎细胞浸润。模型组7个有炎性表现的间盘中,5个浸润的炎症细胞LCA染色阳性,6个浸润的炎症细胞KP-1染色阳性(图3、4)。3个无炎性表现的间盘中,LCA及KP-1染色均为阴性。假手术组与正常组LCA及KP-1染色均为阴性。说明颈性心肌缺血家兔颈椎间盘周围组织存在炎症反应,炎症浸润细胞多为单核一巨噬细胞及淋巴细胞。
实验四颈性心肌缺血家兔颈椎间盘中bFGF和TGF-β1的表达情况
1材料和方法
造模后12周取C6、7颈椎间盘,进行HE染色和免疫组化染色,观察颈性心肌缺血家兔椎间盘中bFGF和TGF-β1的表达情况。
2实验结果及分析
模型组共取得10个间盘。中央部位均为髓核组织及纤维软骨组织,HE染色显示标本软骨有增生、灶状变性,均未见血管组织及炎症细胞;边缘部位为透明或纤维软骨组织,HE染色显示存在灶状增生、粘液变性。其中,7个(70%)间盘周围组织有炎症反应;3个(30%)间盘周围组织未见血管增生及炎细胞浸润等炎症表现。假手术组与正常组HE染色均未见血管增生及炎细胞浸润。模型组颈椎间盘TGF-β1和bFGF及其受体呈阳性表达,尤其在边缘部位,越往中央部位,生长因子及其受体表达越弱。正常组和假手术组的颈椎间盘无论是纤维环和髓核组织内都未见TGF-β1和bFGF及其受体表达。它们促进了颈椎间盘细胞的分化、增殖和细胞外基质的大量合成,这可能是颈椎椎间盘退变的重要原因之一。
实验结论
1采用破坏脊柱三柱结构及颈椎内外平衡的方法,通过颈椎影像学发现的椎体退行性改变及心电图检查出现的缺血性ST-T改变证实,可成功建立颈性心绞痛动物模型。实验结果表明,颈性心绞痛发病机制是颈椎结构中的交感神经纤维受累,使交感神经兴奋,植物神经功能失调,而致冠状动脉痉挛,心肌供血不足并同时影响了房室传导的电激动过程。这是导致心脏功能出现异常的原因之一。
2颈椎病交感神经受刺激、椎基底动脉供血不足时,NPY、ET、CGRP及ANP等神经肽类物质的相应变化可能也共同参与了颈性心绞痛的发病过程。
3颈性心肌缺血家兔颈椎间盘周围组织存在炎症反应,颈椎失稳可能是间盘周围组织产生炎症反应的原因之一;炎症反应可影响到交感神经;慢性炎症因素可能是引起颈性心绞痛发病的重要原因之一。
4 bFGF及TGF-β1促进了颈椎间盘细胞的分化、增殖和细胞外基质的大量合成,这可能是颈椎椎间盘退变的重要原因之一。同时,退变的颈椎间盘本身具有致炎性,炎细胞分泌炎性介质和细胞因子,这些炎性介质作用于交感神经则可能引起支配心脏的交感神经受累,导致颈性心绞痛的发生。颈椎间盘的退变与炎症因素相互影响,对颈性心绞痛的发病都有着密不可分的联系。
The studies were divided two part.
PartⅠ: the Scientific Experiences by Shuchun Sun in Soft-tissue Injury
Shuchun Sun, the chief researcher, is the celebrated orthopedics and traumatology expertise in China. As an instructor of TCM academic experience, a leading scholar in the study of orthopedics and traumatology, a leading figure in academic science of orthopedics and traumatology, Mr. Sun has being engaged in the clinic, scientific research and also the teaching of the orthopedics and traumatology in recent forty years. He is good at using the combined traditional and Western Medicine to diagnose and heal the soft-tissue injury. He advocated the treatment based on the technique of the Traditional Chinese Medicine, carrying forward the characteristic of our homeland traditional medicine. Now, the following is a brief about Mr. Sun's scientific ideas to the soft-tissue injury.
1. Sun's theory in Soft-tissue Injury
Sun's treatment of soft-tissue injury originated from ShangSiYuan of Qing Dynasty's Palace. It is refined from his own skills, widely absorbing achievements from Western Medicine. Sun's unique soft-tissue injury treatment theories include: theory equals to clinical practice; the differentiation of both disease and syndrome; internal treatment supplied with external treatment; dynamics combined with statics, mainly on dynamics; soft-tissue injury differentiation, qi-blood as main points; initiate standardization and develop the superiority.
Mr. Sun always teaches students that attention to patients is not enough. Treating them by our hands is more important. The more we get in touch with patients, the better our hands feel the illness. Only in this way, can we really put theories into clinic treatment.
Mr. Sun also stresses that patients are our best teachers. Good communications with patients help us to improve our technique, which is not taught in books. In his opinion, theory and clinic treatment are of equal importance and should be put together. He makes himself a good example for us.
Mr. Sun believes that symptoms always come along with illness, and we can never separate one from the other. In clinic treatment, what kind of formula and proper technique to choose should be based on an overall analysis of both the illness and signs?
The combination of the disease differentiation in Western Medicine and the syndrome differentiation in TCM is just a model in the clinic therapy. So we should base ourselves upon TCM's theory of treatment according to syndromes, in the meanwhile, utilize the theory and means of examination of the modern medicine reasonably to broaden our vision of treatment, and also to observe and analyze the inherent cause of disease, pathogenesis and evolvement regulation.
Mr. Sun emphasized that both parts and whole must be taken into account when treatment is administered because where there is a sign, there is a cause. Considering the fact that partial damage is the main part in the soft-tissue injury, during the treatment, not only the part but the whole must be focused on. Then, choose corresponding acupoints and proper manipulation under the guidance of either part or whole in order to release hypertonic and stop pain.
What's more, Mr. Sun pointed out that dynamics and statics are opposite but balancing. They support each other and prove a dialectical relationship. The dynamic function is to make the injured limbs get resorted, for the impairment of injured tissues, and for the recovery of the limbs' function.
Mr. Sun also believed the treatment differentiation should be mainly on the qi-blood syndrome differentiation. He thought that qi-blood circulated in the whole body, existing everywhere. Any internal injury, from up and down, skin and flesh, sinew and bone, five Zang-organs and six Fu-organs, wherever it is, qi-blood always comes to be the first affected.
The manipulation in Traditional Chinese Medicine should be based on a mature and standard set of operational rules, in which Mr. Sun is going to call on his peers to make efforts. Although there will be a lot of difficulties in the way, the rules represent the developing direction of this technology, even the cause of Traditional Chinese Medicine.
2. Sun's Dialectics System in Soft-tissue Injury
Based on the diagnosis of inspection, auscultation and olfaction, interrogation and pulse-taking, Mr. Sun takes advantage of his own abundant clinical experiences to refine a set of special diagnosis system which is suitable for the soft-tissue injury.
Interrogation is indispensable in the diagnosis process, and it takes a very important place. Every generation of the medicine expertise pays great attention to interrogation from ancient time to our modem society. We can get the information that the patients felt by themselves, when and where they got injured, the process of their treatments, their health situations, and even the disease history of their families.
Inspection contains the observation of the vitality, color and figure, particularly percept the part of the soft-tissue injury.
Auscultation and olfaction include listening to the patient's words, breath, dyspnea and cough. In addition, listening to the sound from the injured areas is the key point in Sun's theory of soft-tissue injury.
Palpating plays an important role in Sun's soft-tissue treatment. It's different from pulse-taking and is a method to touch and palpate the part or the whole body by the doctor's hands. It can provide the important diagnosis evidence. Palpation provides a thorough understanding of the injured part and has great significance when X-ray is unavailable.
3. Sun's Scientific Characteristic in Soft-tissue Injury
Mr. Sun's manipulation is primarily originated from Shoushan Liu, an old famous TCM doctor. He succeeds to inherit Mr. Liu's manipulation for treating soft-tissue injury. At the mean time, Mr. Sun applied modem anatomical physiology and pathological physiology to organize them into order. On the basis of keeping the curative effect, he also simplified and refined the manipulation, made it easier to master and use. Gradually, with special features, Sun's treatment for soft-tissue injury formed. Summarizing forty years clinic experiences and learning widely from others, Mr. Sun managed to form his special treatment, including the treatment for soft-tissue injury (especially for the cervical vertebra), special treatment for certain disease and function exercise.
Mr. Sun maintained that the curative effect of the manipulation is all by itself, as mentioned in the Medical Principles, Highlights of the Core Techniques of Ortho-correction. Hand indeed has flesh and blood. The fine use of it is felt by stretching. The proper positioning, speed and strength of hand can sense the stagnation of qi-blood, break and contraction of bones and muscles, gall of flesh, and emotional sufferings. Manipulation must live up to the point that "once face symptoms, bodily conditions and the injured parts are clear in mind." Mr. Sun pointed out that the soft tissue sustains weak strengths and is unbearable to strong ones. Thus manipulation can be called "hand skill" only when the patient cannot feel the pain when a method is being applied.
And he also pointed out that application of the manipulation emphasizes on the proper strength that can heal or alleviate the patient's symptoms without pain during the therapy. What is original in Sun's manipulation is being slight, skillful, gentle and soft. PartⅡ: Experimental studies of pathogenesis of Cervical Coronary Heart Disease
Objective:
Along with the quick rhythm of modem society, the change of work manner, increasing opportunity of cervical vertebra bent, incidence of cervical spondylosis (CS)and diseases due to cervical vertebra degeneration rise continuously, now, have become a kind of modem disease accompaning with modem society. At present, domestic and international medical field have noticed that causes of cervical disease can stimulate or oppression nerve root, spinal cord, vertebral artery or sympathetic nerve to produce clinical various syndrome including visceral symptom. Now, the research to causes of cervical diseases has become a new task of world medical field.
In numerous clinical syndrome induced by causes of cervical diseases, cervical coronary heart disease (CCHD)is relatively common. Epidemic investigation show that incidence of CCHD in that of CS take 13%. The cause of CCHD now is considered that cervical degeneration and instability resulting from cervical injary or strain stimulated sympathetic never of predominating cardiovascular function. Now, causes of CCHD still have many aspects to be resolved, how to be in trouble for sympathetic never in causes of CCHD?, is nerve factor involved in only or complex nerve-body fluid mechanism participated in simultaneously? In three cervical ganglion(CG), which is in trouble is more easy to arouse symptom of coronary heart disease(CHD) and appearance of corresponding electrocardiogram expression? If vegetative never functional disturbance due to sympathetic nerve stimulated by cervical degeneration or instability can influence cardiac contractility. These problems have not been had systematic research. Whether myocardial ischemia (MI) induced by CS can lead to organic myocardial damage, and whether CS is one of the causes of CHD still have not unambiguous viewpoint. In basic research, for lack of CCHD animal model, thorough research of its detailed pathogenesis is especially affected.
CS can arise symptom of similar CHD. That CS and CHD come on at the same time is also not rare. According to Dates ,18.4%~53.6% in CS patients have heart disease at the same time. Not knowing the connecton between two diseases is necessary or accidental yet.. Therefore, it is significant to study pathogenesis of CCHD and relation between CS and cardiac function.
Professor Sun Shu-chun thinks that cervical vertebra is related closely with the qi and blood, meridian and zang fu of the whole body, the pathological changes of cervical vertebra can involve in correlative organs. Following degeneration of cervical intervertebral discs(CIVD) and cervical vertebra instability, CS can be occurred. Cervical local hyperosteogeny and aseptic inflammation can stimulate cervical sympathetic nerve fiber or vertebral artery, so pathological changes passing through central or peripheral reflection and nerve-body fluid-blood circulation factor can influence heart and its blood vessel, which result in CCHD. Professor Sun speculated that sympathetic nerve factor maybe took important role in pathogenesis. Professor Sun also thought that CCHD should belong to" tendon off groove and bone in staggering" in pathogenesis of traditional Chinese medicine (TCM) and by using massage, making "bone reset and tendon go back to groove" could alleviate the stimulation of sympathetic nerve and increase blood supply of vertebra artery. thus, corresponding symptom get been treated better. His experience riched single sympathetic effect theory in pathogenesis of CCHD, and put forward that nerve-body fluid- blood circulation factor took important role in appearance of CCHD.
According to "tendon off groove and bone in staggering" theory of TCM, "three columns" theory of modem medicine about spine and theory of internal and external balance of vertebral steady, the experiment made animal cervical vertebra be in staggering and instability with operation, thus induceing MI imitating CHD in electrocardiogram(ECG). The X-rays changes of cervical vertebra and ECG changes were observed regularly to demonstrate if animal model of CCHD was established successfully, the experiment measured changes of neuropeptide substance such as Neuropeptide Y(NPY), Endothelin (ET), Calcitonin gene-related peptide(CGRP), Atrial natriuretic peptide (ANP) in plasma. In order to study that changes of nerve-body fluid-blood circulation induced by cervical instability, vertebra degeneration and sympathetic nerve stimulated is important cause of MI, weaken myocardial contractility and myocardial damage. Author discussed pathogenesis of CCHD, which supplyed academic foundation of prevention and cureing of this disease, at the same time, also riched connotation of diseases related with cervical vertebra. It had been proved by studies on pathogenesis of lumbar disc protrusion that the compression of spinal nerve roots was not the single factor leading to radicular pain and dysfunction and inflammation may be more important to cause impairment on the structure and function of nerve roots. What are the roles of inflammation, inflammatory cytokines and growth factors in the episode of cervical spondyosis? Therefore the cervical intervertebral discs of 3 groups had undergone histological observation for inflammation and detection for the expression of basic fibroblast growth factor (bFGF) and transforming growth factor-β1(TGF-β1) in contrast to the functional status of spinal cord to discuss the role of inflammation, inflammatory cytokines and growth factors in the degeneration of cervical intervertebral discs and impairment of spinal cord. So the pathogenesis of cervical degenerative diseases can be illuminated more completely and it can expand new ideas in the clinical prevention and cure.
Methods:
1.Study of establishment of animal model of myocardial ischemia induced by cervical instability and pathogenesis of CCHD.
30 rabbits were divided into 3 groups randomly, each group of 10, A: normal group; B: model group; C: pseuoperation group. Making CCHD animal model with cutting off supraspinal ligament of C5~6, C6~7, C7~T1, peeling off paravertebral muscle, spining and staggering C6~7 vertebral bodies. After 4, 8 and 12 weeks, taking postero-anterior and lateral radiograph of cervical vertebra to observe changes of cervical intervertebral space, cervical vertebra curvature, intervertebral foramen and hyperplasia aroud vertebra, and analyseing these changes. Furthermore, recording and analyseing changes of ECG before making animal model and 1, 2, 4, 6, 8, 10, 12 weeks after making animal model to observe whether cervical instability lead to ECG of MI.The changes of ECG mainly observed were waveform of each wave, P-R and Q-T interval, ST segment elevation or depression, time of QRS wave, form and direction of T-wave. When finishing the experiment, NPY, ET, CGRP and ANP were measured with radio-immunity methods,
2. Study of the experiment measured changes of neuropeptide substance.
the experiment measured changes of neuropeptide substance such as Neuropeptide Y(NPY), Endothelin (ET), Calcitonin gene-related peptide(CGRP), Atrial natriuretic peptide (ANP) in plasma. In order to study that changes of nerve-body fluid-blood circulation induced by cervical instability, vertebra degeneration and sympathetic nerve stimulated is important cause of MI, weaken myocardial contractility and myocardial damage.
3. study of the infiltration of inflammatory cells.
All discs were divided into underwent fixation, dehydration, paraff'm soak, embedding, paraffin blocks making to detect the expression of LCA and KP-1 by immunohistoChemistry (SABC method) on paraffin sections and hematoxylin-eosin(H-E) staining for histoogical observation to study the infiltration of inflammatory cells. The result was positive when there were yellow or brown granules in the cellular plasm of intervertebral disc cells.
4. study of the infiltration of growth factors.
All discs were divided into underwent fixation, dehydration, paraffin soak, embedding, paraffin blocks making to detect the expression of bFGF and TGF-β1 by immunohistochemistry(SABC method) on paraffin sections and hematoxylin-eosin(H-E) staining for histoogical observation to study the infiltration of inflammatory cells. The result was positive when there were yellow or brown granules in the cellular plasm of intervertebral disc cells.
Results:
1.Changes and analysis of X-rays of cervical vertebra
Integral value of X-rays of model group was obviously higher than that of normal group and pseuoperation group at 4 weeks and discrepancy was significant(P<0.01). Along with time prolonging, the discrepancy is more notable, especially at 12 weeks. Integral value of X-rays of normal group compared with pseuoperation group did not have significant discrepancy(P>0.05). Showing that cervical vertebra has occurred changes of degeneration obviously after cervical vertebra instability caused by operation, and these changes aggravated gradually with times.
2.Changes of ECG and analysis
In experimental period, P-wave time of all animals were measured, but outcome did not have significant discrepancy between every groups(P>0.05). Showing that cervical vertebra instability did not influence electrical exciting course for intra-atrial conduction. After cervical vertebra instability, P-R interval of model group was prolonged, having significant discrepancy (P<0.05), which was compared with before instability. P-R interval of model group which compared with that of normal group and pseuoperation group was prolonged (P<0.05), implying that after cervical vertebra instability, the electrical exciting course of atrioventricular conduction was affected and time from atrium to ventricle was prolonged. Time of QRS-wave of model group which compared with that of normal group and pseuoperation group was prolonged(P<0.05), assoon as cervical vertebra instability. After that time, this instance came into being continuously. Especially, after 8 weeks, discrepancy was more significant(P<0.01). Showing that the electrical exciting course of intraventricular conduction was affected. Q-T interval of model group which compared with that of normal group and pseuoperation group was prolonged (P<0.05), showing that cervical vertebra instability could influence electrical exciting course of intraventricular depolarization and repolarization. ST segment depression, T-wave depression and inversion could been observed in all model animals, after cervical vertebra instability. Along with prolongation of time of cervical vertebra instability, the changes of ST-T existed continuously in model animals, but all animals in normal group and pseuoperation group did not express changes of ST-T all the while. Showing that myocardial ischemia of rabbits could been caused by cervical vertebra instability and staggering. Animal model of CCHD was made successfully.
3. Changes of plasma neuropeptide and analysis
The plasma NPY, ET, ANP of model group was higher than that of normal group and pseuoperation group, and discrepancy was significant (P<0.05,P<0.01). The plasma NPY, ET, ANP between normal group and pseuoperation group did not have significant discrepancy(P>0.05). The plasma CGRP of model group was lower than that of normal group and pseuoperation group, and discrepancy was significant (P<0.05,P<0.01). The plasma CGRP between normal group and pseuoperation group did not have significant discrepancy(P>0.05). Showing that cervical vertebra instability stimulated nerve fiber, which resulted in sympathetic excitability increased. At the same time, synthesis and release of NPY which coexisted with NA in sympathetic system were increased. NPY could contract blood vessel intensively, which made coronary artery spasm and brought about MI. Catecholamine(CA) which was released because of exciting sympathetic nerve, could directly promote the synthesis of vascular endotheliocyte and secretion of ET. Furthermore, plenty of CA could also make vertebrobasilar artery insufficiency(VBI) and brain ischemia, which could stimulate vascular endotheliocyte of brain and expression of ET-mRNA of cerebral neurogliocyte. thus, secretion of ET may be increased. At the same time, cervical vertebra instability and staggering could oppress vertebral artery, which also made ET increased. ET could made coronary artery spasm and brought about MI, as effected of NPY. When release of plasma NPY was increased, NPY could reinforce the regulation of cardial sensory nerve and decrease composition and release of CGRP. Decreasion of CGRP caused by effect of presynatic regulation of NPY could influence normal chronotrpic and inotropic action of heart and the function of expanding coronary artery, which influenced cardiac function and resulted in MI. CA also could make ischemia and anoxia of lung and myocardium, contraction of lung blood vessel and all capillary, returned blood volume increased, pressure of right atrium increased, which resulted in lots of ANP secreted.The changes of ANP might be protective and compensatory reaction of body.
4. Changes of inflammatory cells.
In 10 herniatied cervical intervertebral discs, 7(70%) had infiltration of abundant inflammatory cells in margin of the herniated discs. Most of the inflammatory cells were monocytes and macrophages and there was a small quantity of lymphocytes and granulocytes. There were no infiltration in the normal discs. It was showed by ststistical analysis the incidences of inflammatory cells infiltration in model group were more than normal group and there was statistical difference among these 3 groups.
5. Changes of inflammatory cells.
bFGF and TGF-β1 were negative in most intervertebral discs of normal group and positive in most intervertebra discs of model group. The positive rates of bFGF and TGF-β1 in model gaoup were all significantly more than that in normal group. There was significant difference of the positive, rates of bFGF and TGF-β1 among trauma group, normal group and pseuoperation group.
Conclusion:
1. Confirmed by X-rays of cervical vertebra and ST-T changes of ECG, methods of damaging three column structure of spine and making internal and external imbalance of cervical vertebra could successfully establish animal model ofCCHD.
2.Pathogenesis of CCHD was that cervical sympathetic nerve was simulated, especially ICG and postganglionic fibers. Vegetative never functional disturbance due to stimulating sympathetic nerve could make coronary artery spasm and MI, and also influence electrical exciting course of atrioventricular and intraventricular conduction.
3.Furthermore. When VBI caused by stimulating sympathetic nerve occurred, NPY, ET, CGRP and ANP maybe participated in pathogenesis of CCHD.
4.It hand been proved that nucleus pulposus had the ability to cause inflammation. More than half of degenerative and injured cervical intervertebral discs had inflammation. Therefore inflammation may play an important role in cervica interverbral disc degeneration, pathogenesis and development of cervical spondylosis. What's more, this study showed that chondrocytes in the degenerative intervertebral discs have inflammatory characteristics which could secret inflammatory cytokines. These inflammatory cytokines not only act on the disc cells and influence the matrix synthesis and degradation of intervertebral discs, but also act on spinal cord and nerve roots to cause inflammation, pain and neurotoxic effect which aggravate the nerve impairment of cervical spondylosis patients. It was showed that the expression level of bFGF and TGF-β1 correspond with different phases maybe the important factor inducing the degeneration of cervical intervertebral discs.
引文
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