本文選題：哮喘 + 肺組織��； 參考：《桂林醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:探討Dll4-Notch3信號(hào)對(duì)哮喘大鼠氣道炎癥、氣道重塑的影響。方法:選用SPF級(jí)雌性SD大鼠,霧化吸入OVA及用Notch3、Dll4兩種抗體在體干預(yù)制作哮喘模型。哮喘模型成功后,測(cè)定每組大鼠氣道阻力;常規(guī)HE染色運(yùn)用圖像分析軟件分析氣道壁的嗜酸性粒細(xì)胞(EOS)浸潤情況;測(cè)定相關(guān)指標(biāo)觀察氣道壁厚度(支氣管管腔內(nèi)周長、管壁面積、支氣管平滑肌面積及平滑肌細(xì)胞核數(shù));測(cè)定氣道平滑肌向氣道上皮基底膜遷移情況;免疫組織化學(xué)方法(IHC)觀察各組大鼠氣道壁Notch3、Dll4的表達(dá)情況;采用實(shí)時(shí)熒光定量PCR(RT-qPCR)、蛋白免疫印跡法(Western-blot)檢測(cè)各組大鼠肺組織Notch3、Dll4 m RNA及蛋白的表達(dá)變化。結(jié)果:(1)(1)哮喘1周組與哮喘4周組大鼠的氣道反應(yīng)性、氣道壁EOS浸潤數(shù)量、氣道壁壁厚相關(guān)指標(biāo)都較正常對(duì)照組顯著增高(P0.05),哮喘4周組較哮喘1周組上述指標(biāo)顯著增高(P0.05),且隨致敏天數(shù)延長上述指標(biāo)逐漸增高(P0.05)。(2)哮喘1周組與哮喘4周組相對(duì)于正常對(duì)照組,平滑肌距基底膜的距離減小(P0.05),哮喘4周組與哮喘1周組相比距離減小(P0.05)。(3)Notch3、Dll4在哮喘1周組、哮喘4周組的mRNA表達(dá)水平均高于正常對(duì)照組(P0.05),且此兩組間有顯著性差異(P0.05)。Western-blot及IHC所測(cè)結(jié)果基本同RT-qPCR結(jié)果一致。(2)(1)哮喘1周組、哮喘4周組的氣道反應(yīng)性、氣道壁EOS浸潤數(shù)量、氣道壁壁厚相關(guān)指標(biāo)高于正常對(duì)照組(P0.05),哮喘4周組的上述指標(biāo)亦高于哮喘1周組(P0.05)。Notch3治療1周組、Notch3治療4周組的上述指標(biāo)均高于正常對(duì)照組(P0.05),Notch3治療4周組的上述指標(biāo)高于Notch3治療1周組(P0.05)。Dll4治療1周組、Dll4治療4周組的上述指標(biāo)均高于正常對(duì)照組(P0.05),Dll4治療4周組的上述指標(biāo)高于Dll4治療1周組(P0.05)。哮喘4周組上述指標(biāo)高于Notch3治療1周組和Notch3治療4周組(P0.05),哮喘4周組上述指標(biāo)高于Dll4治療4周組,無組間對(duì)比意義(P0.05)。(2)哮喘1周組、哮喘4周組的支氣管平滑肌距基底膜的距離較正常對(duì)照組減小(P0.05),哮喘4周組較哮喘1周組上述指標(biāo)減小(P0.05)。Notch3治療1周組、Notch3治療4周組的以上指標(biāo)較正常對(duì)照組減小(P0.05),Notch3治療4周組的以上指標(biāo)較Notch3治療1周組減小(P0.05)。Dll4治療1周組、Dll4治療4周組的以上指標(biāo)較正常對(duì)照組減小(P0.05),Dll4治療4周組的以上指標(biāo)較Dll4治療1周組減小(P0.05)。哮喘4周組上述指標(biāo)小于Notch3治療1周組和Notch3治療4周組(P0.05),哮喘4周組上述指標(biāo)大于Dll4治療4周組,無組間對(duì)比意義(P0.05)。(3)哮喘1周組、哮喘4周組Notch3、Dll4 mRNA表達(dá)水平均高于正常對(duì)照組(P0.05),哮喘4周組的Notch3、Dll4表達(dá)水平高于哮喘1周組(P0.05)。Notch3治療1周組、Notch3治療4周組的以上指標(biāo)均高于正常對(duì)照組(P0.05),Notch3治療4周組的以上指標(biāo)高于Notch3治療1周組(P0.05)。Dll4治療1周組、Dll4治療4周組的以上指標(biāo)均高于正常對(duì)照組(P0.05),Dll4治療4周組的以上指標(biāo)高于Dll4治療1周組(P0.05)。哮喘4周組上述指標(biāo)高于Notch3治療1周組和Notch3治療4周組(P0.05),哮喘4周組上述指標(biāo)高于Dll4治療4周組,無組間對(duì)比意義(P0.05)。Western-blot及IHC所測(cè)結(jié)果基本同RT-qPCR結(jié)果一致。結(jié)論:Dll4在哮喘大鼠氣道上皮細(xì)胞有特異性表達(dá),Notch3在整個(gè)哮喘肺組織中呈高表達(dá)。抑制Dll4-Notch3信號(hào)可改善哮喘氣道炎癥及氣道重構(gòu)。表明Dll4-Notch3信號(hào)參與了哮喘的發(fā)病。
[Abstract]:Objective: To investigate the effect of Dll4-Notch3 signal on airway inflammation and airway remodeling in asthmatic rats. Methods: using SPF grade female SD rats, inhaling OVA and using Notch3, Dll4 two kinds of antibodies to make asthma model in vivo. After the asthma model was successful, the airway resistance was measured in each group of rats; the airway wall was analyzed by the routine HE staining software and the image analysis software was used to analyze the airway wall. The infiltration of eosinophil (EOS), the thickness of the airway wall (perimeter of the bronchus, the area of the tube wall, the area of the bronchial smooth muscle and the number of the nucleus of the smooth muscle of the bronchial smooth muscle), the migration of the airway smooth muscle to the basal membrane of the airway epithelium, and the expression of Notch3 and Dll4 in the airway wall of the rats were observed by the immunohistochemical method (IHC). The changes in the expression of Notch3, Dll4 m RNA and protein in lung tissues of each group were detected by real time fluorescence quantitative PCR (RT-qPCR) and protein immunoblotting (Western-blot). Results: (1) (1) the airway responsiveness of the 1 week asthma group and the 4 week group of asthma, the number of infiltration of the airway wall EOS and the wall thickness of the airway were significantly higher than that of the normal control group. High (P0.05), 4 weeks asthma group was significantly higher than that of the 1 week asthma group (P0.05), and the above index increased with the number of sensitization days (P0.05). (2) the distance between the 1 week asthma group and the 4 week asthma group compared with the normal control group, the distance of the smooth muscle from the basilar membrane decreased (P0.05), the 4 week asthma group was compared with the 1 week group of asthma (P0.05). (3) Notch3, Dll4 In the 1 week asthma group, the expression level of mRNA in the 4 week asthma group was higher than that in the normal control group (P0.05), and there was a significant difference between the two groups (P0.05) the results of.Western-blot and IHC were basically the same as the results of RT-qPCR. (1) the 1 weeks of asthma, the airway reactivity of the 4 week group of asthma, the number of EOS infiltration in the airway wall and the index of the wall thickness of the airway were higher than that of the normal. In the control group (P0.05), the above indexes in the 4 week asthma group were also higher than those in the 1 week group of asthma (P0.05) for 1 weeks, and the above indexes in the 4 week group were higher than those in the normal control group (P0.05). The above indexes in the 4 week group of Notch3 treatment were higher than those in the 1 week group of Notch3 (P0.05) for 1 weeks, and the above indexes in the 4 weeks group were all higher than those of the normal control. Group (P0.05), the above index in the 4 week group of Dll4 treatment was higher than that of the group of Dll4 for 1 weeks (P0.05). The above index in the 4 week group of asthma was higher than that in the 1 week group of Notch3 and 4 weeks in the Notch3 treatment group (P0.05). The above index in the 4 week group of asthma was higher than that in the 4 week group of Dll4 (P0.05). (2) the bronchial smooth muscle of the bronchial smooth muscle from the 1 week group of asthma and the 4 weeks of asthma was from the basilar membrane. The distance from the normal control group decreased (P0.05), the 4 week asthma group was less than that of the 1 week asthma group (P0.05) in the 1 week group, and the above index in the 4 week group decreased (P0.05). The above index in the 4 week group of Notch3 treatment was lower than that in the 1 week group of Notch3 (P0.05).Dll4 therapy 1 weeks, and Dll4 was used to treat the above indexes in the 4 week group. Compared with the normal control group (P0.05), the above index in the 4 week group of Dll4 treatment was less than that of the 1 week group (P0.05). The above indexes in the 4 week group of asthma were less than that of the 1 weeks group and the 4 week group (P0.05), the 4 week asthma group was more than the 4 week group with Dll4 treatment, and there was no comparison between the groups (P0.05). (3) the asthmatic 1 weeks group, 4 weeks of asthma, Notch3, Dll4 M The expression level of RNA was higher than that of the normal control group (P0.05). The expression of Notch3 and Dll4 in the 4 week asthma group was higher than that in the 1 week group of asthma (P0.05).Notch3 treatment 1 weeks, and the above indexes in the 4 week group of Notch3 treatment were higher than that of the normal control group (P0.05). The above index in the 4 week group of Notch3 treatment was higher than that of the 1 week group of Notch3 treatment (P0.05) for 1 weeks, and 4 weeks for the treatment. The above index in the group was higher than that of the normal control group (P0.05). The above index in the 4 week group of Dll4 treatment was higher than that of the 1 week group (P0.05). The above indexes in the 4 week group of asthma were higher than that in the 1 week group of Notch3 and the 4 week group of Notch3 (P0.05). The above indexes in the 4 week group of asthma were higher than those in the 4 week group with Dll4, and there was no comparison significance (P0.05).Western-blot and IHC. The results are basically the same as the results of RT-qPCR. Conclusion: Dll4 has a specific expression in the airway epithelial cells of asthmatic rats, and Notch3 is highly expressed in the lung tissue of the whole asthma. The inhibition of Dll4-Notch3 signal can improve the airway inflammation and airway remodeling in asthma. It indicates that the Dll4-Notch3 signal is involved in the onset of asthma.

【學(xué)位授予單位】：桂林醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R562.25

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