脾气虚证患者唾液淀粉酶活性及其相关指标的研究

发布时间：2018-01-16 23:20

本文关键词：脾气虚证患者唾液淀粉酶活性及其相关指标的研究　出处：《广州中医药大学》2016年硕士论文　论文类型：学位论文

【摘要】：研究背景：既往研究根据“脾主涎”等中医理论发现脾气虚证患者唾液淀粉酶(salivary alpha-amylase, sAA)活性比值(酸刺激后／酸刺激前)较健康者明显下降,该指标1993年被卫生部颁布的《中药新药临床研究指导原则》列入脾气虚证疗效评价的参考指标,是为数不多的得到政府部门认可的证候微观指标。但其在多种疾病脾气虚证患者中的检出率约为60%-70%,特异性不够高,表明单一使用该指标作为脾气虚证的微观指标的信息还不够全面。目的：以sAA活性改变为切入点,从同病异证和异病同证角度探讨脾气虚证患者sAA活性及其相关指标(唾液pH值、唾液流率、总蛋白及离子浓度等)改变情况,进一步丰富中医“脾主涎”的科学内涵,并观察脾气虚证患者sAA活性及相关指标合参对临床检出率的影响。方法：一、运用EP管自然流出法、口中转动棉柱法和口中咀嚼棉柱法分别采集55例健康志愿者酸刺激前、后的唾液,检测并比较sAA活性及总活性、唾液流率指标变化情况,为筛选较优的唾液采集方法提供依据。二、采集广州中医药大学第一附属医院脾胃病科门诊及住院部101例慢性浅表性胃炎(chronic superficial gastritis, CSG)和60例重症肌无力(myasthenia gravis,MG)患者酸刺激前和酸刺激后的唾液,CSG患者纳入的证型主要为脾气虚、脾虚湿热和肝胃不和3个证型(分别为36、40和25例),上午9：00～-11：00采集；MG患者纳入的证型主要为脾气虚、脾虚湿热和脾肾两虚3个证型(分别为32、23和5例),晨醒30 min内采集。三、招募筛选青中年健康志愿者并采集其酸刺激前和酸刺激后的唾液,采集时间分晨醒30 min内和上午9：00～11：00(早餐1h后)。四、检测受试者sAA活性、唾液流率、pH值、总蛋白浓度、Ca2+及Cl-浓度。sAA活性采用Bernfeld法,唾液流率采用称重法计算,pH值使用pH计检测,总蛋白浓度采用BCA蛋白定量试剂盒检测,Ca2+及C1-浓度采用离子定量试剂盒检测。五、与健康对照组比较sAA活性及其相关指标的改变情况,尝试对sAA活性及相关指标进行多指标合参并评价其临床检出率情况。结果：一、唾液样品采集方法的优化：EP管自然流取法与口中转动棉柱法的sAA活性及总活性和唾液流率在酸刺激后均升高,反映了健康者唾液腺对酸的正常应激能力,且两者在酸刺激后各指标升高的检出率较为接近,其中又以口中转动棉柱法有结果客观、可平行采集足量唾液的优势；而口中咀嚼棉柱法的结果与前两种方法差异较大,各指标比值升高的检出率明显低于前两者,未能客观反映健康者酸刺激后sAA活性升高的现象。因而选择口中转动棉柱法作为后续研究唾液样品的采集方法。二、脾气虚证患者与健康者唾液指标检测：(一)CSG患者与健康者比较：(1)酸刺激后,健康者sAA活性、sAA总活性、流率和pH值升高,反映了健康者对酸刺激的正常应激能力；唾液总蛋白浓度酸刺激后降低,可能是酸刺激后唾液流率升高,总蛋白浓度相对降低所致。(2)CSG及其脾气虚证和脾虚湿热患者酸刺激后sAA活性降低,并且sAA活性比值低于健康者,说明CSG及其脾虚患者唾液腺对酸的应激能力降低,再次重复验证既往研究结果。肝胃不和患者sAA活性及总活性比值高于脾气虚证和脾虚湿热证患者,与健康者无明显差异；唾液流率、pH值、总蛋白浓度、Ca2+及C1-浓度在证型间无明显差异；说明sAA活性改变为脾虚患者相对特异性指征。(3)酸刺激前、后,CSG及其三证型患者唾液流率均低于健康者,总蛋白浓度、Ca2+及Cl-浓度均高于健康者,说明患者唾液成分浓度相对于健康者已发生改变,提示可能与患者植物神经系统功能紊乱有关。(4)酸刺激后脾气虚证患者唾液pH值降低的例数(16例)高于健康者(2例),说明脾气虚证患者唾液缓冲能力降低。(二)MG患者与健康者比较：(1)MG及其脾气虚证患者酸刺激后sAA活性降低,并且sAA活性比值和sAA总活性比值均低于健康者,说明MG及其脾虚患者唾液腺对酸的应激能力降低,再次重复验证既往研究结果。MG脾气虚证患者sAA活性改变与CSG脾气虚证患者表现类似,说明sAA活性改变为脾气虚证患者普遍出现的现象。MG及其脾气虚证和脾虚湿热患者酸刺激前sAA活性高于健康者,提示在基础状态下MG脾虚患者交感神经偏亢。 (2)MG及其脾气虚证患者唾液流率酸刺激前后均低于健康者,总蛋白浓度、Ca2+及C1-浓度高于健康者,说明患者唾液成分浓度相对于健康者已发生改变,再次提示可能与患者植物神经系统功能紊乱有关。(3)酸刺激后,脾气虚证患者唾液pH值降低例数(17例)多于健康者(6例),pH值比值低于健康者,说明脾气虚证患者唾液缓冲能力降低,这与CSG脾气虚证患者表现类似,而MG脾气虚证患者出现率更高,可能与MG病理改变相对更严重有关。(4)在脾气虚证和脾虚湿热证之间各指标均无差异,提示可能是MG患者以脾气虚弱为本,所以证型在指标上未表现明显差异。(5)CSG与MG脾气虚证患者sAA活性比值均较健康者降低,重复证实了sAA活性比值降低可能是脾气虚证患者的相对共性表现,目前结果表明,两种疾病脾气虚证患者的唾液分泌可能存在相似的病理生理改变。结论：口中转动棉柱法采集唾液样品简便、可靠,可供后期研究者继续选用。本研究根据中医理论“脾主涎”证实了CSG和MG脾气虚证患者不仅仅表现在sAA活性异常这个单一的指标上,还表现在sAA总活性、唾液流率、pH值、总蛋白浓度、Ca2+和Cl-浓度这些指标改变上,为丰富“脾主涎”内涵提供了参考。脾气虚证和脾虚湿热患者可能均存在植物神经系统紊乱和唾液腺对酸的应激能力降低的病理表现。除了sAA活性比值,唾液pH值比值改变对于脾气虚证可能也有一定参考价值。将sAA活性、唾液流率和pH值进行合参后在脾气虚证的检出率相较以往单用sAA活性指标提高了约11.00%,但由于本研究的例数有限,多指标合参的结果尚有待后续进一步验证。
[Abstract]:Background: Previous studies based on "spleen controlling saliva and saliva in patients with deficiency of TCM theory that temper amylase (salivary alpha-amylase, sAA) activity ratio (after acid stimulation / acid before stimulation) decreased significantly compared with the healthy people, the reference index in the 1993 by the Ministry of Health issued the" Chinese medicine clinical research guiding principles > included in evaluation of temper syndrome curative effect, is one of the few to get syndrome micro index recognition. But the government departments in a variety of diseases in patients with spleen deficiency syndrome, the detection rate is about 60%-70%, the specificity is not high enough, that single use of the index as microscopic indexes of spleen qi deficiency of the information is not comprehensive. Objective: to change the activity of sAA as the starting point, from the same angle with different syndromes syndrome and different disease activity and sAA in patients with deficiency of spleen index (pH value of saliva, salivary flow rate, total protein and ion concentration changes), To further enrich the scientific connotation of TCM "spleen controlling saliva", and to observe the activity of sAA in patients with deficiency of temper and related indicators and impact on the clinical detection rate. Methods: first, using EP tube natural outflow method, in turn chewing cotton fibre cotton column method and mouth were collected in 55 healthy volunteers acid after stimulation, saliva, detect and compare the activity of sAA and the total activity, changes of salivary flow rate index, provide the basis for screening the optimal saliva collection method. Two, collecting the First Affiliated Hospital of Guangzhou University of Chinese Medicine Department of outpatient and inpatient department and 101 cases of chronic superficial gastritis (chronic superficial, gastritis, CSG) and 60 cases of severe myasthenia gravis (myasthenia gravis, MG) patients before and after acid stimulation acid stimulated saliva syndrome CSG patients included mainly spleen deficiency, spleen deficiency and dampness heat and liver stomach disharmony syndrome (3 cases of 36,40 and 25 respectively), morning 9:00 - -1 1:00 collection; syndrome MG patients included mainly spleen deficiency, spleen deficiency and dampness heat and spleen kidney two empty syndrome type 3 (32,23 and 5), collected in the morning wake up 30 min. Three, the recruitment of young and middle-aged healthy volunteers were collected and screened the acid stimulation before and after acid stimulation of saliva collection time divided into morning wake up 30 min and 9:00 (1h to 11:00 morning after breakfast. Four), were determined by sAA activity, salivary flow rate, pH value, total protein concentration, Ca2+ concentration and Cl- activity of.SAA by Bernfeld method, the salivary flow rate by weight method calculation, pH value using the pH tester, total protein concentration. Detected by BCA protein assay kit, Ca2+ and C1- by ion concentration Quantification Kit detection. Five, change of control group sAA activity and the related indicators and health, try on the activity of sAA and related indicators of multi index parameters and to evaluate its clinical detection rate. Results: a saliva sample Optimization of acquisition method: EP tube natural flow method and rotation cotton column method in sAA activity and total activity and salivary flow rate were increased after acid stimulation, reflects the health of the normal salivary acid stress, and both increased in each index after acid stimulation rate is close to that of the and with the rotation of cotton column method is objective, parallel to capture enough saliva advantage; difference chew cotton column method with the former two methods greatly, each index ratio increased the detection rate was significantly lower than the former two, failed to reflect the healthy subjects after acid stimulation increased the activity of sAA and select the phenomenon. In turn the cotton column method as the collection method of follow-up study. Two saliva samples, detection of spleen qi deficiency patients and healthy subjects: (a) saliva index between CSG patients and healthy subjects: (1) after acid stimulation, healthy sAA activity, sAA activity, flow rate and P The increase of H value, reflects the health of the normal stress ability of acid stimulation; salivary total protein concentration decreased after acid stimulation, salivary flow rate may be increased after acid stimulation, total protein concentration is relatively reduced. (2) reduce the activity of sAA and CSG in spleen qi deficiency and spleen damp heat patients after acid stimulation, and sAA activity were lower than health, CSG and salivary glands of patients with spleen deficiency acid stress reduced ability to repeat past results. Verify the disharmony between liver and stomach in patients with sAA activity and total activity ratio was higher than that in patients with spleen deficiency syndrome and spleen deficiency and dampness heat syndrome, with no significant difference between healthy subjects; salivary flow rate, pH value, total protein concentration, Ca2+ had no significant difference. And the concentration of C1- in syndrome type; sAA activity change for patients with spleen deficiency relative specific indications. (3) after acid stimulation, CSG, and three patients were lower than that of normal salivary flow rate, total protein concentration, Ca2+ and Cl- The concentration was higher than that of healthy subjects, that saliva concentration patients compared to healthy have been changed, suggesting that it may be related to dysfunction of autonomic nervous system. (4) after acid stimulation of saliva in patients with deficiency of temper pH decreased the number of cases (16 cases) was higher than that of healthy subjects (2 cases), indicating a reduced capacity of saliva in patients with deficiency of buffer temper. (two) MG patients in comparison with healthy subjects: (1) reduce the activity of sAA and MG in patients with spleen deficiency syndrome after acid stimulation, and the activity of sAA and the ratio of the total activity of sAA was significantly lower than that of healthy subjects, MG and salivary gland in patients with spleen deficiency to acid stress reduced ability to repeat previous research results verify.MG temper the changes of activity of sAA and CSG in patients with syndrome of deficiency of spleen qi deficiency patients is similar to that of sAA activity is a common phenomenon in patients with spleen deficiency syndrome and spleen deficiency syndrome and.MG patients with spleen deficiency and dampness heat before acid stimulation of sAA activity is higher than that of healthy subjects Based on the state of MG, suggesting that spleen deficiency patients with sympathetic hyperactivity. (2) MG before and after acid stimulation in patients with spleen deficiency syndrome, salivary flow rate was lower than that of healthy subjects, the total protein concentration, Ca2+ and concentration of C1- was higher than that of healthy subjects, patients compared to healthy subjects saliva concentration has changed again, suggest that may be related with plant the disorder of the nervous system. (3) after acid stimulation, temper saliva in patients with deficiency of pH decreased the number of cases (17 cases) than healthy subjects (6 cases), pH ratio value is lower than that of healthy subjects, reduce the capacity of saliva in patients with deficiency of spleen and CSG buffer, the patients with spleen deficiency syndrome were similar, while MG appeared in patients with spleen deficiency syndrome at higher rates, and MG may be the pathological changes more serious. (4) between spleen deficiency syndrome and spleen deficiency syndrome of dampness heat of each index showed no difference, suggesting that MG patients with spleen weakness in the syndrome type, so in the index showed no significant difference Different. (5) sAA activity ratio of CSG and MG in patients with deficiency of spleen were lower than healthy subjects decreased, repeated that the lower sAA activity ratio may be relatively common symptoms of spleen deficiency syndrome patients, the results showed that two kinds of diseases in patients with spleen deficiency syndrome, the secretion of saliva may have a similar pathophysiological change. Conclusion: in rotation cotton column method to collect saliva samples is simple and reliable, can be used for later researchers to select the research. According to TCM theory of "spleen saliva" was confirmed by CSG and MG in patients with spleen deficiency syndrome, not only in the sAA abnormal activity of this single index, but also in the total activity of sAA, salivary flow rate, pH value, total protein concentration Ca2+ and Cl-, changing the concentration of these indicators, provide a reference for the rich connotation of "spleen controlling saliva". In patients with spleen deficiency syndrome and spleen deficiency and dampness heat may exist in autonomic nervous system disorders and salivary gland to acid stress ability decreased Pathology. In addition to the sAA activity ratio, the ratio of salivary pH change in spleen deficiency may also have certain reference value. The activity of sAA, salivary flow rate and pH of ginseng in spleen qi deficiency detection rate compared to the previous single sAA activity index increased by about 11%, but due to the limited number of research cases. Multi index combination results remains to be further verified.

【学位授予单位】：广州中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R228

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