平喘固本合劑對哮喘小鼠氣道慢性炎癥及重塑的影響及機制
發(fā)布時間：2018-10-26 20:44
【摘要】：目的建立小鼠哮喘氣道重塑模型；初步探討平喘固本合劑對小鼠哮喘模型的氣道慢性炎癥及重塑的作用及相關(guān)機制,為防治哮喘氣道重塑提供新的研究依據(jù)。 方法健康雌性昆明系小鼠50只隨機分為五組,每組10只,正常對照組(A)、哮喘組(B)、平喘固本合劑干預組(C)、布地奈德霧化干預組(D)、聯(lián)合治療干預(E)。B、C、D、E組小鼠采用卵清白蛋白(OVA)和氫氧化鋁粉的復合制劑致敏、OVA霧化激發(fā)建立哮喘小鼠模型,并分別給予C、D、E組不同干預治療。對各組支氣管肺泡灌洗液(BALF)中各種細胞進行分類并計數(shù),肺組織病理切片行HE染色后觀察氣道重塑情況并且采用Image-Pro PLus6.0病理圖像分析軟件測定氣道管壁厚度及管壁面積,采用免疫組織化學法檢測核因子-κB(NF-κB)、血管內(nèi)皮轉(zhuǎn)化因子(VEGF)的表達水平。數(shù)據(jù)采用ANOVA檢測進行組間分析,LSD檢驗進行組內(nèi)兩兩比較。 結(jié)果①A組小鼠無喘息癥狀；B組小鼠喘息癥狀明顯加重,且皮毛暗淡；C、D、E組有喘息癥狀,但均較B組輕；②HE染色顯示所有藥物干預組較單純哮喘模型組炎癥細胞浸潤、平滑肌肥厚及黏膜氣道組織水腫、上皮細胞脫落等表現(xiàn)明顯減輕。其中以D和E組最為明顯；③B組BALF中細胞總數(shù),嗜酸性粒細胞、中性粒細胞、淋巴細胞比例均高于其他各組,差別有統(tǒng)計學意義(P0.01)；A組低于其它各組,差別亦有統(tǒng)計學意義(P0.01)；C組與D組小鼠BALF中嗜酸性粒細胞數(shù)無明顯統(tǒng)計學差異(P0.01)與哮喘B組相比明顯降低,差異有統(tǒng)計學意義(P0.01)④圖像分析結(jié)果顯示,Wat/Pi(μ2/μm)各組氣道管壁的厚度分別為(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組顯著低于B組、C組、D組、E組,差異有統(tǒng)計學意義(P分別等于0.000、0.000、0.000、0.012)；C組高于D、E組,差異具有統(tǒng)計學意義(P分別等于0.009,0.000)；D組與E組無明顯統(tǒng)計學意義(P=0.193)；B組顯著高于A組、C組、D組和E組差異有統(tǒng)計學意義(P0.000)。⑤小■肺組織免疫組化結(jié)果顯示,小鼠肺組織中VEGF陽性表達主要在氣管粘膜上皮、支氣管平滑肌、血管內(nèi)皮及血管平滑肌,炎性細胞(主要是嗜酸性粒細胞)也有表達。平均光密度(IOD)測定：分別為(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組、C組、D組、E組中的VEGF光密度值均明顯高于A組,差異具有統(tǒng)計學意義(P0.01)；B組的光密度值明顯高于C組、D組、E組,差異具有統(tǒng)計學意義(P0.01)；D組的光密度值低于C組,差異具有統(tǒng)計學意義(P=0.036)；E組的光密度值明顯低于C組、D組,差異具有統(tǒng)計學意義(P0.01)。NF-κB主要表達在氣道粘膜上皮細胞胞質(zhì)內(nèi),分別為(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組、C組、D組的平均光密度值均明顯高于A組,差異具有統(tǒng)計學意義(P0.01)；C組與D組、E組與A組無明顯統(tǒng)計學差異(P=0.53,0.76)；B組的平均光密度值明顯高于C組、D組、E組,差異具有統(tǒng)計學意義(P=0.042,0.000,0.000)。 結(jié)論運用OVA致敏及霧化的方法可以復制出慢性哮喘氣道重塑的小鼠模型；哮喘時NF-κB、VEGF的表達水平均提高,可能有加重氣道慢性炎癥反應(yīng)及引起氣道結(jié)構(gòu)改變的作用,參與氣道重塑的發(fā)生；平喘固本合劑可以抑制氣道炎癥細胞的浸潤,下調(diào)哮喘模型小鼠氣道組織中NF-κB和VEGF的表達,達到抑制哮喘氣道慢性炎癥及重塑的作用。
[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.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R725.6
本文編號：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.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R725.6
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