支气管哮喘证候分布规律研究及哮喘平冲剂对哮喘模型大鼠气道重塑的影响
本文选题：支气管哮喘 切入点：哮喘平冲剂 出处：《安徽中医药大学》2016年硕士论文
【摘要】：1.临床研究1.1目的:对142例支气管哮喘(bronchial asthma,BA)患者进行中医证候学调查,探求哮喘中医证候分布规律,为哮喘的中医辨证分型和临床治疗提供理论依据。1.2方法:参考资料制定哮喘证候表,结合哮喘控制测试(ACT)评分及哮喘控制情况表,对符合条件的142例哮喘患者的资料利用统计软件对相关因素进行分析,总结哮喘中医证型分布规律。1.3结果:142例哮喘患者中寒哮证共有65例(所占比例45.77%),哮喘易感体质以阳虚质和痰湿质为主,分别占比35.91%和32.39%,说明哮喘的发病与体质因素相关。2.实验研究1.1目的:观察哮喘平冲剂对哮喘模型大鼠细胞因子IL-17、Notch信号通路的影响,探讨哮喘平冲剂治疗哮喘的免疫机制及分子机制。1.2方法:将84只健康雄性的SD大鼠随机分为7组:正常对照组、哮喘模型组、泼尼松西药组、金水宝中成药组,哮喘平冲剂低、中、高剂量组,每组12只,常规喂养7天后开始造模。致敏2次,以卵蛋白(Ovalbumin,OVA)喷雾激发,每日1次,共激发5周。雾化造模结束后中药低、中、高剂量组予以哮喘平冲剂、西药组予以泼尼松、中成药组予以金水宝胶囊灌胃治疗,正常组、模型组予以等量生理盐水灌胃,每天一次,连续2周。其中,中药高剂量组予以0.625g/kg/d为给药剂量(相当于成人的12.5倍),中药中剂量组予以0.313g/kg/d为给药剂量(相当于成人的6.25倍),中药低剂量组予以0.157g/kg/d为给药剂量(相当于成人的3.13倍),西药组予以0.313g/kg/d为给药剂量,中成药组予以0.313g/kg/d为给药剂量。两周后取材检测血清及肺泡灌洗液IL-17的含量,免疫组化检测Notch信号通路Notch1-4的表达,探讨哮喘平冲剂对哮喘治疗的机制。1.3结果:1.3.1大鼠血清IL-17含量:与正常组相比,模型组大鼠血清IL-17含量显著高于正常组(P0.01)。与模型组相比,各治疗组大鼠血清IL-17含量均降低(P0.01)。西药组与中药高剂量组相比,差异无统计学意义(P0.05)。中成药组与中药低剂量组相比,差异无统计学意义(P0.05)。中药中剂量组、中药高剂量组大鼠血清IL-17含量均低于中成药组(P0.05)。1.3.2大鼠肺泡灌洗液IL-17含量:与正常组相比,模型组大鼠肺泡肺泡灌洗液IL-17含量显著高于正常组(P0.01)。与模型组相比,各治疗组大鼠肺泡肺泡灌洗液IL-17含量均降低(P0.01)。西药组与中药高剂量组相比,差异无统计学意义(P0.05)。中成药组与中药低剂量组相比,差异无统计学意义(P0.05)。中药中剂量组、中药高剂量组大鼠肺泡灌洗液IL-17含量均低于中成药组(P0.05)。1.3.3免疫组化检测各组大鼠肺组织Notch1-4表达:与正常组相比,模型组肺组织Notch1、Notch2平均光密度值显著高于正常组(P0.01);与模型组相比,治疗组各组肺组织Notch1、Notch2平均光密度值均降低(P0.01);中药高剂量组与西药组比较,差异无统计学意义(P0.0 5),中药中剂量与中药低剂量比较,Notch1、Notch2平均光密度值有所降低(P0.05);中药低剂量与中成药组比较,差异无统计学意义(P0.0 5)。与正常组比较,模型组Notch3平均光密度值有增高(P0.05),其余各组肺组织中Notch3平均光密度值均明显减弱,且两组间比较无明显差异(P0.0 5)。Notch4平均光密度值在各组中均无明显增高。1.3.4 RT-PCR检测肺组织Notch1-4 mRNA表达:与正常组相比较,模型组肺组织Notch1、Notch2 m RNA表达显著高于正常组(P0.0 1),与模型组相比较,治疗组各组肺组织Notch1、Notch2 m RNA表达均降低(P0.0 5),中药高剂量组与西药组比较,差异无统计学意义(P0.05),中药中剂量组Notch1、Notch2 m RNA表达较中药低剂量组降低(P0.0 5),中药低剂量组与中成药组比较,差异无统计学意义(P0.0 5);Notch3 m RNA在模型组及对照组各组肺组织中表达均减弱,两组比较无差异(P0.0 5)。Notch4m RNA在模型组肺组织及对照组肺组织中均表达不明显。3.结论:3.1在所研究的142例哮喘患者中证型以哮喘冷哮证居多。3.2哮喘平冲剂治疗哮喘的免疫机制可能是抑制白细胞介素-17(IL-17)的产生,从而减轻气道炎症;治疗哮喘的分子机制可能是通过下调Notch信号通路中参与T细胞分化与激活的Notch1、Notch2的表达,从而延缓哮喘的气道重塑。
[Abstract]:1. clinical research 1.1 Objective: 142 cases of bronchial asthma (bronchial asthma, BA) in patients with TCM syndrome of asthma survey, explore the distribution regularity of TCM syndromes, and provide a theoretical basis for the.1.2 method for traditional Chinese medicine syndrome type and clinical treatment of asthma: reference for asthma syndrome scale combined with asthma control test (ACT) score and asthma control table, to meet the conditions of 142 cases of patients with asthma data using statistical software to analyze the related factors, summed up the law.1.3 results of TCM syndrome of asthma distribution: 142 asthma patients in cold syndrome were found in 65 cases (accounted for 45.77%), asthma susceptible to yang deficiency and phlegm dampness, accounted for more than 35.91% and 32.39%, indicating the onset of.2. and physical experimental study on related factors of asthma 1.1 Objective: To observe the asthma Granule on asthmatic cytokines in rats with IL-17, the effect of Notch signaling on asthma The immune mechanism and the molecular mechanism of.1.2 granule in treating asthma asthma methods: 84 healthy male SD rats were randomly divided into 7 groups: normal control group, asthma model group, prednisone group, western medicine, Chinese medicine group, Jinshuibao xiaochuanping powder, low, high dose group, 12 rats in each group, routine feeding 7 days after modeling. Sensitized with ovalbumin 2 times (Ovalbumin, OVA) spray excitation, 1 times daily for 5 weeks. Total excitation is low, traditional Chinese medicine atomization after the modeling, the high dose group of xiaochuanping powder, western medicine group was given prednisone, Chinese medicine group was given Jinshuibao capsule gavage treatment and the normal group, model group were given normal saline, once a day for 2 weeks. Among them, high dose of Chinese medicine group were given 0.625g/kg/d dosage (equivalent to 12.5 times the adult), Chinese medicine dose group received 0.313g/kg/d dose (equivalent to 6.25 times a person), Chinese medicine low the 0.157g dose group /kg/d dose (equivalent to 3.13 times the adult), western medicine group received 0.313g/kg/d dose, Chinese medicine group received 0.313g/kg/d dose after two weeks. The content of serum and bronchoalveolar lavage fluid IL-17 detection, immunohistochemical detection of Notch signal pathway in the expression of Notch1-4, to investigate the xiaochuanping powder on the results the mechanism of.1.3 treatment of asthma: the content of IL-17 in serum of 1.3.1 rats: compared with normal group, the content of IL-17 in serum of rats in model group were significantly higher than the normal group (P0.01). Compared with the model group, the content of IL-17 in serum of rats in each treatment group were decreased (P0.01). Compared with the traditional Chinese medicine Western medicine group and high dose group, no significant difference meaning (P0.05). Compared with the traditional Chinese medicine group low dose group, the difference was not statistically significant (P0.05). Chinese medicine dose group, high dose of Chinese medicine group rats serum IL-17 levels were lower than the traditional Chinese medicine group (P0.05).1.3.2 in rat alveolar lavage fluid containing IL-17 Weight: compared with the normal group, model group of rat alveolar alveolar lavage fluid IL-17 was significantly higher than normal group (P0.01). Compared with the model group, the alveolar alveolar lavage fluid of rats in each treatment group IL-17 decreased (P0.01). Compared with the traditional Chinese medicine Western medicine group and high dose group, no significant difference (P0.05) compared with traditional Chinese medicine. Chinese medicine group, low dose group, the difference was not statistically significant (P0.05). Chinese medicine dose group, high dose of Chinese medicine group rat bronchoalveolar lavage fluid IL-17 levels were lower than the traditional Chinese medicine group (P0.05) immunohistochemical detection of.1.3.3 in lung tissue of rats Notch1-4 expression: compared with normal group, model group, lung tissue Notch1 Notch2, the average optical density value is significantly higher than the normal group (P0.01); compared with the model group, treatment group Notch1 lung tissue, the average optical density values of Notch2 were decreased (P0.01); high dose of Chinese medicine group and Western medicine group, the difference was not statistically significant (P0.0 5), Chinese Medicine In comparison, Chinese medicine dose and low dose of Notch1, the average optical density values of Notch2 decreased (P0.05); relatively low dose of traditional Chinese medicine and traditional Chinese medicine group, the difference was not statistically significant (P0.0 5). Compared with the normal group, Notch3 model group, the average optical density increased (P0.05), Notch3 in other groups in the lung tissues of the average light the density values were significantly decreased, and no significant difference between the two groups (P0.0 5) the average optical density values of.Notch4 in each group were not significantly higher expression of Notch1-4 mRNA in lung tissue of.1.3.4 RT-PCR: compared with the normal group, model group, lung tissue Notch1, Notch2 expression of M RNA was significantly higher than the normal group (P0.0 1). Compared with the model group, treatment group Notch1 lung tissue, the expression of Notch2 m and RNA were decreased (P0.0 5), high dose of Chinese medicine group and Western medicine group, the difference was not statistically significant (P0.05), Chinese medicine dose group Notch1, Notch2 m RNA expression compared with the low dose of Chinese medicine group Reduce (P0.0 5), compared with the low-dose group of Chinese medicine and traditional Chinese medicine group, the difference was not statistically significant (P0.0 5); the expression of Notch3 m RNA in the model group and control group in lung tissue were decreased, the two groups have no difference (P0.0 5).Notch4m expression of RNA was not obvious.3. in lung tissue of model group and conclusion the control group in the lung tissues: 3.1 immune mechanism in 142 asthmatic patients in the syndrome type of asthma with cold asthma are.3.2 xiaochuanping powder for the treatment of asthma may be the inhibition of interleukin -17 (IL-17) generation, so as to reduce airway inflammation; molecular mechanism of treating asthma may down regulate the expression of Notch1 T cell differentiation and activation of the Notch signaling pathway involved in the expression of Notch2, thus delaying the airway remodeling of asthma.
【学位授予单位】：安徽中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R256.12
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