过敏体质嗜酸性粒细胞相关因子表达及过敏康调体干预机制研究

发布时间：2018-03-10 17:12

本文选题：过敏康　切入点：过敏体质　出处：《北京中医药大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:寻找过敏体质与平和体质之间的差异性因子,确立过敏体质的特异性分子标记;观察中药"过敏康"通过调理过敏体质治疗过敏性疾病的临床效果;探索中药"过敏康"通过调理过敏体质治疗过敏性疾病的作用机制与嗜酸性粒细胞功能调控的相关性,并寻找该机制的作用靶点,以期明确过敏体质的微观作用机制,为过敏体质理念更好的应用于临床提供指导。方法:本研究通过收集门诊患者及张贴招募海报的方式获取受试者,并依据中医体质分类判定标准和过敏性鼻炎诊断标准对受试者进行筛选,最终纳入受试者52人,男女比例为1:1,年龄在18到40岁之间。其中过敏体质32例,平和体质20例,分别命名为过敏体质组、平和体质组。对过敏体质组受试者给予中药"过敏康"干预三个月,平和体质组作为对照,不采取干预措施。为实验描述方便,将治疗后的过敏体质组命名为干预后过敏体质组,并在后续实验中对每组受试者的血液样本分别进行蛋白芯片检测、ELISA检测、QGP检测。三组受试者均在清晨空腹采集血样,用普通采血管抽取受试者静脉血20ml,离心后得到血清,运用蛋白芯片技术检测受试者外周血血清样本中IL-5、GM-CSF、IL-3、eotaxin、RANTES、IL-4、IL-13、IL-9 蛋白水平,运用 ELISA 试剂盒检测受试者外周血血清样本中IL-25、IL-33、TLSP、ECP蛋白水平;用乙二胺四乙酸抗凝管(EDTA抗凝管)抽取受试者静脉血5ml,取36ul全血按照QuantiGene Sample Processing Kit说明书预处理,以QuantiGene Plex法检测受试者外周血中IL-5、eotaxin、CCR3mRNA表达水平。数据处理:采用spss22.0软件处理,平和体质者与过敏体质者的指标差异选用独立t检验方法,过敏体质干预前后的指标变化选用配对t检验方法,数据均以X±s表示,P0.05为有统计学差异。结果:1.中药"过敏康"通过调理过敏体质的方式治疗过敏性鼻炎受试者,经3个月的干预周期后,患者的TNSS、TNNSS、RQLQ评分表的各项分值均有下降,其主要评价指标即TNSS评分表中的鼻塞、鼻痒、流涕、喷嚏这四个鼻部症状在治疗第4至8周改善最为明显,并在之后的4周疗程中持续下降。在试验结束一年后的回访中,可发现患者在服药后的第二个过敏季节,过敏症状和生活状态较试验结束时均有显著改善,且部分患者没有复发。2.过敏体质受试者在治疗后血清总IgE及相应过敏原的特异性IgE均有下降,但不显著。3.过敏体质与平和体质的差异性因子:两组相比,过敏体质组受试者外周血中 IL-5、GM-CSF、eotaxin、RANTES、IL-4、IL-13、IL-9、IL-25、IL-33、TLSP、ECP蛋白水平及IL-5、eotaxin、CCR3mRNA水平高于平和体质组受试者,差异具有统计学意义(P0.05);过敏体质组受试者外周血中IL-3蛋白水平高于平和体质组受试者,差异无统计学意义(P0.05)。4.给予"过敏康"干预3个月后,过敏体质组受试者外周血中 IL-5、eotaxin、RANTES、IL-4、IL-13、IL-25、TLSP、ECP 蛋白水平及IL-5、eotaxin、CCR3mRNA水平较干预前降低,差异具有统计学意义(P0.05),过敏体质组受试者外周血中GM-CSF、IL-9、IL-33蛋白水平较干预前有降低,但差异没有统计学意义(p0.05)。结论:1.中药"过敏康"通过调理过敏体质可有效改善过敏性鼻炎患者的临床症状和生活质量,且这种改善具有持续性。2.IL-5、GM-CSF、eotaxin、RANTES、IL-4、IL-13、IL-9、IL-25、IL-33、TLSP、ECP、CCR3是平和体质与过敏体质之间的差异性因子。3.中药"过敏康"调理过敏体质的作用机制涉及对嗜酸性粒细胞相关功能的调控,IL-5、eotaxin、RANTES、IL-4、IL-13、IL-25、TLSP、ECP、CCR3 可能为"过敏康"调理过敏体质的作用靶点。
[Abstract]:Objective: to find out the differences between the allergic constitution factor with mild constitution, the establishment of specific molecular markers of allergy; to observe the clinical effect of Guominkang by conditioning allergy treatment of allergic diseases; explore traditional Chinese medicine "Guominkang" through the correlation mechanism of conditioning allergy treatment of allergic diseases and eosinophil function the regulation, and look for the targets of the mechanism of the micro mechanism in order to clear the allergy, allergic constitution concept better application in clinical guidance. Methods: This study by collecting outpatient and post recruitment poster access to subjects, and on the basis of TCM constitution classification standards and the allergic rhinitis diagnostic criteria of subjects were screened, the final 52 subjects, male to female ratio was 1:1, at the age of 18 to 40 years old. 32 cases of allergic constitution, and constitution of 20 An example, named allergy group, and physical group. In allergic subjects treated with traditional Chinese medicine "Guominkang" intervention for three months, and constitution as control group without intervention. Description is convenient for the experiment, the allergy group after treatment of allergies named after the intervention group. And in the follow-up experiment subjects blood samples in each group respectively, protein chip detection, ELISA detection, QGP detection. Three groups of subjects were in the early morning fasting blood samples were collected, using ordinary blood drawn venous blood 20ml, serum was obtained after centrifugation, detected by protein chip technology in IL-5, serum samples of week the blood of subjects in GM-CSF, IL-3, eotaxin, RANTES, IL-4, IL-13, IL-9 protein level by ELISA assay IL-25, peripheral blood serum samples of subjects in IL-33, TLSP, ECP protein level; with EDTA anticoagulant tubes (EDTA tubes) by extraction Try to take venous blood 5ml, 36ul blood QuantiGene Sample Processing Kit in accordance with the pretreatment instructions, detected by QuantiGene Plex method by IL-5, peripheral blood eotaxin, the expression level of CCR3mRNA. Data processing: using spss22.0 software, using independent t test method and index difference constitution and allergy, index the change before and after the intervention with allergy paired t test method, the data were expressed as X + s, P0.05 is statistically significant. Results: 1. Chinese medicine "Guominkang" by conditioning the allergy treatment of allergic rhinitis subjects after 3 months after the intervention period, patients with TNSS, TNNSS, RQLQ score the scores were decreased, the main evaluation index TNSS score of nasal congestion, nasal itching, sneezing, runny nose, nasal symptoms of the four improvement in the treatment of fourth to 8 weeks is the most obvious, and continued in the 4 weeks after the treatment under Drop. A year after the end of follow-up in the trial, patients can be found in second after taking a seasonal allergies, allergic symptoms and living condition is improved significantly at the end of the test, and some patients with no recurrence of.2. allergic subjects after treatment serum total IgE and allergen specific IgE were decreased however, no significant differences in factor.3. allergy with mild Constitution: compared to the two groups, allergic subjects IL-5, peripheral blood GM-CSF, eotaxin, RANTES, IL-4, IL-13, IL-9, IL-25, IL-33, TLSP, ECP protein level and IL-5, eotaxin, CCR3mRNA levels higher than normal constitution group of subjects, the difference was statistically significant (P0.05); allergic subjects level of IL-3 protein in the peripheral blood level and physical subjects, there was no statistically significant difference (P0.05.4.) to "Guominkang" after 3 months of intervention, allergic subjects in the peripheral blood of I L-5, eotaxin, RANTES, IL-4, IL-13, IL-25, TLSP, ECP protein level and IL-5, eotaxin, reduce the level of CCR3mRNA compared with before intervention, the difference was statistically significant (P0.05), allergic subjects GM-CSF, peripheral blood IL-9, IL-33 protein levels are lower than those before intervention, but no differences statistical significance (P0.05). Conclusion: 1. Chinese medicine "Guominkang" by conditioning allergy in patients with allergic rhinitis can effectively improve the clinical symptoms and quality of life, and has sustained.2.IL-5, GM-CSF, eotaxin, RANTES, IL-4, IL-13, IL-9, IL-25, IL-33, TLSP, ECP CCR3 is the improvement mechanism of Chinese herbal medicine "the difference between flat and.3. factor of physique and allergy Guominkang" allergy conditioning are involved in regulating eosinophil function, the related IL-5, eotaxin, RANTES, IL-4, IL-13, IL-25, TLSP, ECP, CCR3 might be the targets of conditioning allergy Guominkang. "

【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R259

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