补肾平喘方对大鼠哮喘模型的作用及机制的实验初探

发布时间：2018-05-17 20:01

本文选题：补肾平喘方 + 哮喘　；参考：《蚌埠医学院》2014年硕士论文

【摘要】：目的：通过观察补肾平喘方（Bushen Pingchuan Prescription）对大鼠哮喘模型的防治作用，初步探讨本方抗哮喘的作用及作用机制，为该处方的进一步开发提供初步的实验依据。方法： SPF级清洁雄性SD大鼠60只，采用腹腔注射卵蛋白（OVA）与氢氧化铝致敏、OVA雾化吸入激发来复制哮喘动物模型。第1，7天除空白对照组外，其余大鼠腹腔注射卵蛋白（OVA）与氢氧化铝致敏，之后将其随机分为5组：模型组（等容量蒸馏水）、补肾防喘片组(6.21g/kg)、补肾平喘方高剂量组(49.68g/kg)、补肾平喘方中剂量组(24.84g/kg)、补肾平喘方低剂量组(12.42g/kg)，每组10只。第15天起每天雾化激发，于激发前半小时各给药组给药。第1,2周雾化浓度为1％，第3,4周为1.5％，第5,6周为2％，第7,8周为2.5％。于末次给药激发后，禁食24h，麻醉、取血、取大鼠肺脏；测定肺脏重量、肺脏指数，分离取血清及血浆,采用ELISA法检测血清中IL-4，IFN-γ，IgE，CORT的含量及血浆中ACTH的含量。将肺组织放入10％的甲醛里浸泡，待进行HE染色，在光学显微镜下观察大鼠肺组织形态结构，自动成像系统拍摄图片。免疫组化法检测肺组织内β-AR的含量。结果： 1.补肾平喘方通过补肾温阳、宣肺降气，改善大鼠肾虚哮喘症状如精神萎靡、喘息、咳嗽等。 2.补肾平喘方能显著降低大鼠肺脏指数，与模型空白组相比有显著差异性（P㩳0.01），其中补肾平喘方中剂量组效果明显。 3.补肾平喘方能显著改善哮喘肺组织病理水平，与模型空白组相比具有显著性差异。 4.补肾平喘方能显著降低血清IL-4同时升高血清IFN-γ的水平，与模型对照组相比有显著差异性（P㩳0.05、P㩳0.01），抑制Th2细胞的优势表达，调节哮喘炎性反应。 5.补肾平喘方显著降低大鼠血清中IgE的水平，，与模型对照组相比有显著差异性（P㩳0.05、P㩳0.01）。 6.补肾平喘方能显著降低大鼠血清CORT，血浆ACTH的水平，调节HPA轴的功能，与模型对照组相比，差异具有显著性（P㩳0.01、P㩳0.05）。 7.补肾平喘方能提高大鼠肺组织中β-AR的水平，与模型对照组相比具有显著性差异（P㩳0.05、P㩳0.01）。结论： 1.哮喘模型组大鼠肺脏病理形态显著病变，血清中IL-4升高、IFN-γ降低，IgE显著提高，同时大鼠出现一系列的肾虚症状精神萎靡、喘息、咳嗽等，证明采用卵蛋白加氢氧化铝注射致敏并给予雾化激发可以成功复制哮喘动物模型。 2.补肾平喘方降低大鼠肺脏指数、血清IL-4与IgE，同时升高IFN-γ水平，可能与抑制Th2细胞的优势表达，调节Th1与Th2的平衡有关。 3.补肾平喘方能升高CORT，ACTH的表达，减轻肺组织病变程度，可能与调节HPA轴的平衡有关，即通过下丘脑-腺垂体-肾上腺轴来调节糖皮质激素的水平，发挥免疫抑制及抗炎作用；此外还提高了 AR的水平，通过扩张支气管及扩张血管等作用来减缓哮喘症状。 4.补肾平喘方温补肾阳、宣肺降逆，对哮喘模型大鼠具有一定的防治作用。
[Abstract]:Objective: By observing the preventive and therapeutic effects of Bushen Pingchuan Prescription( Bushen Pingchuan Prescription) on asthma in rats, the effect and mechanism of Bushen Pingchuan Prescriptionof Bushen on asthma in rats were preliminarily discussed, which provided a preliminary experimental basis for the further development of this prescription. Methods: 60 SPF grade clean male SD rats were injected intraperitoneally with ovalbumin and aluminum hydroxide to induce asthma model. On the 1st day of 7 days, the other rats were sensitized with aluminum hydroxide by intraperitoneal injection of ovalbumin OVA, except for the blank control group. Then it was randomly divided into five groups: model group (equal volume distilled water, Bushen anti-asthma tablet group, 6.21 g / kg 路kg ~ (-1), high dose group (49.68 g / kg 路kg ~ (-1), medium dose group (24.84 g / kg) and low dose group (12.42 g / kg 路kg ~ (-1), 10 rats in each group. Every day after 15 days of atomization, each group was administered half an hour before excitation. The atomization concentration was 1 at week 1, 1.5 at week 3, 2 at week 5 and 2 at week 7, and 2. 5 at week 7. The lung weight, lung index, serum and plasma levels of IL-4, IFN- 纬, IgECORT and ACTH in plasma were determined by ELISA method. The lung tissue was immersed in 10% formaldehyde for HE staining. The morphologic structure of lung tissue was observed under optical microscope and the images were taken by automatic imaging system. The content of 尾-AR in lung tissue was detected by immunohistochemical method. Results: 1. Bushen Pingchuan prescription can improve asthmatic symptoms of kidney deficiency such as depression, wheezing and coughing by tonifying kidney and warming yang, promoting lung and reducing qi. 2. Bushen Pingchuan recipe can significantly reduce the lung index of rats, compared with the model blank group, there is a significant difference between the P0. 01, among which the effect of Bushen Pingchuan decoction medium dose group is obvious. 3. Bushen Pingchuan recipe can significantly improve the lung pathological level of asthma compared with the model blank group has a significant difference. 4. Bushen Pingchuan recipe could significantly reduce serum IL-4 and increase the level of serum IFN- 纬. Compared with the model control group, there was a significant difference between the two groups. Compared with the model control group, Bushen Pingchuan recipe could inhibit the predominant expression of Th2 cells and regulate the inflammatory response of asthma. 5. Bushen Pingchuan recipe significantly decreased the level of IgE in serum of rats, and there was significant difference compared with the model control group. 6. Bushen Pingchuan recipe could significantly reduce the level of Cort, plasma ACTH and regulate the function of HPA axis in rats. Compared with the model control group, the difference was significant. 7. Bushen Pingchuan recipe could increase the level of 尾 -AR in lung tissue of rats, and had significant difference compared with the model control group. Conclusion: 1. In the asthmatic model group, the pathological changes of lung were significant, the serum level of IFN- 纬 was significantly increased, and a series of symptoms of kidney deficiency, such as depression, wheezing and coughing, were found in the rats. It was proved that the asthmatic animal model could be successfully induced by injection of ovalbumin and aluminum hydroxide. 2. The decrease of lung index, serum IL-4 and IgE, and the increase of IFN- 纬 level may be related to the inhibition of the dominant expression of Th2 cells and the regulation of the balance between Th1 and Th2. 3. Bushen Pingchuan prescription can increase the expression of Corton-ACTH and reduce the degree of lung pathological changes, which may be related to regulating the balance of HPA axis, that is, regulating the level of glucocorticoid through hypothalamus-pituitary-adrenal axis, and exerting immunosuppressive and anti-inflammatory effects. It also improves the level of AR and relieves asthma symptoms by dilating bronchi and dilating blood vessels. 4. The prescription of tonifying kidney and relieving asthma has preventive and therapeutic effect on asthmatic rats.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R259

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