平喘固本合剂对哮喘小鼠气道慢性炎症及重塑的影响及机制
发布时间：2018-10-26 20:44
【摘要】：目的建立小鼠哮喘气道重塑模型；初步探讨平喘固本合剂对小鼠哮喘模型的气道慢性炎症及重塑的作用及相关机制,为防治哮喘气道重塑提供新的研究依据。 方法健康雌性昆明系小鼠50只随机分为五组,每组10只,正常对照组(A)、哮喘组(B)、平喘固本合剂干预组(C)、布地奈德雾化干预组(D)、联合治疗干预(E)。B、C、D、E组小鼠采用卵清白蛋白(OVA)和氢氧化铝粉的复合制剂致敏、OVA雾化激发建立哮喘小鼠模型,并分别给予C、D、E组不同干预治疗。对各组支气管肺泡灌洗液(BALF)中各种细胞进行分类并计数,肺组织病理切片行HE染色后观察气道重塑情况并且采用Image-Pro PLus6.0病理图像分析软件测定气道管壁厚度及管壁面积,采用免疫组织化学法检测核因子-κB(NF-κB)、血管内皮转化因子(VEGF)的表达水平。数据采用ANOVA检测进行组间分析,LSD检验进行组内两两比较。 结果①A组小鼠无喘息症状；B组小鼠喘息症状明显加重,且皮毛暗淡；C、D、E组有喘息症状,但均较B组轻；②HE染色显示所有药物干预组较单纯哮喘模型组炎症细胞浸润、平滑肌肥厚及黏膜气道组织水肿、上皮细胞脱落等表现明显减轻。其中以D和E组最为明显；③B组BALF中细胞总数,嗜酸性粒细胞、中性粒细胞、淋巴细胞比例均高于其他各组,差别有统计学意义(P0.01)；A组低于其它各组,差别亦有统计学意义(P0.01)；C组与D组小鼠BALF中嗜酸性粒细胞数无明显统计学差异(P0.01)与哮喘B组相比明显降低,差异有统计学意义(P0.01)④图像分析结果显示,Wat/Pi(μ2/μm)各组气道管壁的厚度分别为(6.61±1.14),(16.66±1.52),(11.57±1.26),(9.53±1.93),(8.56±1.35)μm2/μm,F=54.59,P0.01。A组显著低于B组、C组、D组、E组,差异有统计学意义(P分别等于0.000、0.000、0.000、0.012)；C组高于D、E组,差异具有统计学意义(P分别等于0.009,0.000)；D组与E组无明显统计学意义(P=0.193)；B组显著高于A组、C组、D组和E组差异有统计学意义(P0.000)。⑤小■肺组织免疫组化结果显示,小鼠肺组织中VEGF阳性表达主要在气管粘膜上皮、支气管平滑肌、血管内皮及血管平滑肌,炎性细胞(主要是嗜酸性粒细胞)也有表达。平均光密度(IOD)测定：分别为(11.57±1.64,35.87±4.92,28.28±2.02,25.06±2.58,16.31±2.41F=85.45,P0.01),B组、C组、D组、E组中的VEGF光密度值均明显高于A组,差异具有统计学意义(P0.01)；B组的光密度值明显高于C组、D组、E组,差异具有统计学意义(P0.01)；D组的光密度值低于C组,差异具有统计学意义(P=0.036)；E组的光密度值明显低于C组、D组,差异具有统计学意义(P0.01)。NF-κB主要表达在气道粘膜上皮细胞胞质内,分别为(16.52±2.04,20.88±0.78,19.57±1.21,18.32±0.93,17.66±0.78,F=14.73,P0.01)B组、C组、D组的平均光密度值均明显高于A组,差异具有统计学意义(P0.01)；C组与D组、E组与A组无明显统计学差异(P=0.53,0.76)；B组的平均光密度值明显高于C组、D组、E组,差异具有统计学意义(P=0.042,0.000,0.000)。 结论运用OVA致敏及雾化的方法可以复制出慢性哮喘气道重塑的小鼠模型；哮喘时NF-κB、VEGF的表达水平均提高,可能有加重气道慢性炎症反应及引起气道结构改变的作用,参与气道重塑的发生；平喘固本合剂可以抑制气道炎症细胞的浸润,下调哮喘模型小鼠气道组织中NF-κB和VEGF的表达,达到抑制哮喘气道慢性炎症及重塑的作用。
[Abstract]:Objective To establish a model of airway remodeling of asthma in mice, and to explore the role and mechanism of Pingchuan Guben Mixture on airway chronic inflammation and remodeling in mice asthma model, and to provide a new basis for prevention and treatment of asthma airway remodeling. Methods 50 healthy female Kunming mice were randomly divided into five groups: 10 rats in each group, normal control group (A), asthma group (B), antiasthmatic solid mixture intervention group (C), bubulider atomization intervention group (D), combined therapy intervention (E). B, C D, E group mice were sensitive to ovalbumin (OVA) and aluminum hydroxide powder, and the model of asthma mice was established by atomization, and different interventions were given to C, D and E groups, respectively. Treatment. Various cells were classified and counted in each group of bronchoalveolar lavage fluid (BAL). After HE staining of lung tissue pathological section, airway remodeling was observed and image-Pro PLus6.0 pathological image analysis software was used to determine airway tube wall thickness and wall thickness. The expression of vascular endothelial transformation factor (VEGF) in nuclear factor-Sepharose B (NF-Sepharose B) and vascular endothelial transformation factor (VEGF) was detected by immunohistochemical method. Level. The data were analyzed by ANOVA, and the LSD test was performed in two groups within the group. Results There were no wheezing symptoms in Group A mice; the wheezing symptoms in Group B mice were significantly increased and the fur was dim; C, D, E groups had wheezing symptoms but were lighter in Group B; and EHE staining showed that all drug intervention groups were treated with simple asthma model group. The inflammatory cell infiltration, smooth muscle hypertrophy and mucosal airway tissue edema, epithelial cell shedding, etc. The total number of cells, eosinophil, neutrophils and lymphocyte in group B was significantly higher than that in other groups (P <0.01), and the difference was statistically significant (P There was no significant difference in eosinophil number between group C and group D (P0.01). Compared with group D, there was no significant difference in eosinophil count in group C (P0.01), and the difference was statistically significant (P <0.01). The results showed that Wat/ Pi (. 2/. mu.m) had a thickness of (6.61%) in each group. 1. 14), (16. 66, 1. 52), (11. 57, 1. 26), (9. 53, 1. 93), (8. 56, 1. 35). m 2/. mu.m, F = 54. 59, P0. 01. A group was significantly lower than Group B, Group C, Group D, Group E, differences were statistically significant (P was equal to 0. 000, 000, 000, 0. 012); Group C was high There was no significant difference between group D and group E (P = 0.0193); group B was significantly higher than group A, group C, group D and group E (P <0.01). The expression of VEGF in lung tissue of mice is mainly in tracheal mucosa, bronchial smooth muscle, vascular endothelium and vascular smooth muscle, inflammatory cells (mainly eosinophil). The optical density of VEGF in group B, group C, group D and E was significantly higher than that in group A (P0.01), and the optical density of group B was significantly higher than that in group C. The optical density of group D was significantly lower than that in group C (P = 0.01). The value of optical density in group E was significantly lower than that in group C and group D. The average optical density of group C and D was significantly higher than that in group A (P0.01). There was no significant difference between group C and group D, group E and group A (P = 0. 53, 0. 76). The average optical density value of group B was significantly higher than that of group C, group D and group E. Conclusion: The mice model with chronic asthma airway remodeling can be reproduced by using the method of self-induced sensitization and atomization; the expression level of NF-100B and VEGF in asthma can be increased, and it is possible to increase the chronic inflammatory response of the airway and induce the change of airway structure. It can inhibit the infiltration of airway inflammation cells and down-regulate the expression of NF-B-B and VEGF in airway tissues of asthmatic model mice to inhibit the chronic asthma airway.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.6
本文编号：2296896
[Abstract]:Objective To establish a model of airway remodeling of asthma in mice, and to explore the role and mechanism of Pingchuan Guben Mixture on airway chronic inflammation and remodeling in mice asthma model, and to provide a new basis for prevention and treatment of asthma airway remodeling. Methods 50 healthy female Kunming mice were randomly divided into five groups: 10 rats in each group, normal control group (A), asthma group (B), antiasthmatic solid mixture intervention group (C), bubulider atomization intervention group (D), combined therapy intervention (E). B, C D, E group mice were sensitive to ovalbumin (OVA) and aluminum hydroxide powder, and the model of asthma mice was established by atomization, and different interventions were given to C, D and E groups, respectively. Treatment. Various cells were classified and counted in each group of bronchoalveolar lavage fluid (BAL). After HE staining of lung tissue pathological section, airway remodeling was observed and image-Pro PLus6.0 pathological image analysis software was used to determine airway tube wall thickness and wall thickness. The expression of vascular endothelial transformation factor (VEGF) in nuclear factor-Sepharose B (NF-Sepharose B) and vascular endothelial transformation factor (VEGF) was detected by immunohistochemical method. Level. The data were analyzed by ANOVA, and the LSD test was performed in two groups within the group. Results There were no wheezing symptoms in Group A mice; the wheezing symptoms in Group B mice were significantly increased and the fur was dim; C, D, E groups had wheezing symptoms but were lighter in Group B; and EHE staining showed that all drug intervention groups were treated with simple asthma model group. The inflammatory cell infiltration, smooth muscle hypertrophy and mucosal airway tissue edema, epithelial cell shedding, etc. The total number of cells, eosinophil, neutrophils and lymphocyte in group B was significantly higher than that in other groups (P <0.01), and the difference was statistically significant (P There was no significant difference in eosinophil number between group C and group D (P0.01). Compared with group D, there was no significant difference in eosinophil count in group C (P0.01), and the difference was statistically significant (P <0.01). The results showed that Wat/ Pi (. 2/. mu.m) had a thickness of (6.61%) in each group. 1. 14), (16. 66, 1. 52), (11. 57, 1. 26), (9. 53, 1. 93), (8. 56, 1. 35). m 2/. mu.m, F = 54. 59, P0. 01. A group was significantly lower than Group B, Group C, Group D, Group E, differences were statistically significant (P was equal to 0. 000, 000, 000, 0. 012); Group C was high There was no significant difference between group D and group E (P = 0.0193); group B was significantly higher than group A, group C, group D and group E (P <0.01). The expression of VEGF in lung tissue of mice is mainly in tracheal mucosa, bronchial smooth muscle, vascular endothelium and vascular smooth muscle, inflammatory cells (mainly eosinophil). The optical density of VEGF in group B, group C, group D and E was significantly higher than that in group A (P0.01), and the optical density of group B was significantly higher than that in group C. The optical density of group D was significantly lower than that in group C (P = 0.01). The value of optical density in group E was significantly lower than that in group C and group D. The average optical density of group C and D was significantly higher than that in group A (P0.01). There was no significant difference between group C and group D, group E and group A (P = 0. 53, 0. 76). The average optical density value of group B was significantly higher than that of group C, group D and group E. Conclusion: The mice model with chronic asthma airway remodeling can be reproduced by using the method of self-induced sensitization and atomization; the expression level of NF-100B and VEGF in asthma can be increased, and it is possible to increase the chronic inflammatory response of the airway and induce the change of airway structure. It can inhibit the infiltration of airway inflammation cells and down-regulate the expression of NF-B-B and VEGF in airway tissues of asthmatic model mice to inhibit the chronic asthma airway.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.6
【参考文献】
相关期刊论文 前10条
1 林荣军,王为达,潘玉娟,孙秀芳,官如明;青岛市区0～14岁儿童哮喘流行病学调查[J];青岛大学医学院学报;2002年03期
2 于璐;张燕;陈程;崔海福;严兴科;王富春;;针灸对哮喘肺功能影响的临床研究进展[J];环球中医药;2010年02期
3 朱建明;白建文;林闽加;;MMP-9、TIMP-1分别对哮喘小鼠和气道重塑小鼠作用的比较研究[J];临床肺科杂志;2009年06期
4 殷晓峰,冯玉麟,刘春涛;大鼠支气管哮喘模型气道重构的研究[J];四川医学;2002年05期
5 揭志军;金美玲;蔡映云;袁正宏;胡芸文;徐杨;;白介素-4和白介素-13促进人肺成纤维细胞ADAM33基因的表达[J];复旦学报(医学版);2006年03期
6 张金琴;张静;薛琴;;影响哮喘患儿治疗依从性的因素调查及护理干预[J];临床和实验医学杂志;2008年11期
7 韩征利;董翠香;曲政海;刘秀美;周立;;小青龙汤和人参五味子汤二步序贯治疗对哮喘小鼠呼吸道炎症与重塑的影响[J];实用儿科临床杂志;2011年04期
8 刘芝华;;复方甘草酸苷的临床应用[J];天津药学;2007年06期
9 熊瑛;熊彬;王宋平;杨礼腾;李阳轲;孙兴旺;;麻黄水提物对哮喘豚鼠气道炎症和气道上皮细胞STAT1信号传导影响[J];西部医学;2008年05期
10 吕小华;吴铁;覃冬云;;甘草对慢性小鼠哮喘模型气道炎症及外周血Th_1/Th_2失衡的影响[J];中国临床药理学与治疗学;2006年05期
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