發(fā)布時(shí)間：2018-04-20 12:33

  本文選題：維生素D2 + 哮喘　； 參考：《安徽醫(yī)科大學(xué)》2012年碩士論文

【摘要】：目的：哮喘是一種由多種細(xì)胞特別是肥大細(xì)胞、嗜酸性粒細(xì)胞、T淋巴細(xì)胞、嗜中性粒細(xì)胞、氣道上皮細(xì)胞及多種細(xì)胞組分參與的慢性氣道炎癥反應(yīng)性疾病，免疫調(diào)節(jié)在哮喘發(fā)病過(guò)程中有重要作用，維生素D在免疫調(diào)節(jié)中的作用越來(lái)越受到重視，研究發(fā)現(xiàn)維生素D對(duì)Th1/Th2平衡有調(diào)節(jié)作用，本實(shí)驗(yàn)通過(guò)維生素D2對(duì)哮喘大鼠模型進(jìn)行干預(yù)，比較各組試驗(yàn)大鼠肺泡灌洗液中IL-4水平、肺泡灌洗液中細(xì)胞計(jì)數(shù)及分類(lèi)、肺部ICAM-1的表達(dá)，探討維生素D在哮喘大鼠的抗炎作用及其可能機(jī)制。 方法：按照隨機(jī)分組的原則，將30只SD雄性大鼠隨機(jī)分組成3組，分別為哮喘組、維生素D2組、對(duì)照組，每組10只。（1）哮喘組：于第1、8、15天分別給予無(wú)菌腹腔注射10%OVA1ml（內(nèi)含卵蛋白100mg及氫氧化鋁100mg）致敏，自第16天開(kāi)始置于密閉霧化罐中用1%OVA霧化吸入激發(fā)，持續(xù)霧化1周，每天激發(fā)30分鐘致其出現(xiàn)呼吸急促、喘息或呼吸困難。（2）維生素D2(VD2)組：誘喘方法同哮喘組，從第15天開(kāi)始給予腹腔注射維生素D25×10~5U/（kg.d），隔天1次，，共3次。（3）對(duì)照組：以生理鹽水代替OVA和維生素D2。在末次激發(fā)后24小時(shí)內(nèi)處死大鼠，收集肺組織及肺泡灌洗液（BALF）。將BALF離心后分離上清及沉淀細(xì)胞，并將肺組織經(jīng)脫水透明、石蠟包埋等處理后制作成肺標(biāo)本切片。運(yùn)用Elisa方法對(duì)BALF上清中白細(xì)胞介素4（IL-4）水平進(jìn)行測(cè)定，以及用改良牛氏計(jì)數(shù)臺(tái)對(duì)BALF沉淀細(xì)胞進(jìn)行計(jì)數(shù)并將沉淀細(xì)胞涂片染色后進(jìn)行分類(lèi)計(jì)數(shù)。運(yùn)用免疫組織化學(xué)方法技術(shù)測(cè)定大鼠肺組織標(biāo)本細(xì)胞間粘附分子1（ICAM-1）水平。通過(guò)統(tǒng)計(jì)學(xué)方法對(duì)各組指標(biāo)數(shù)據(jù)進(jìn)行分析比較。 結(jié)果：VD2組IL-4含量（501.54±17.96）pg/ml明顯低于哮喘組（615.37±94.77）pg/ml（P0.01）；VD2組IL-4含量與對(duì)照組比較無(wú)統(tǒng)計(jì)學(xué)意義；VD2組大鼠肺泡灌洗液沉渣中細(xì)胞總數(shù)（72.80±24.44）×10~6、中性粒細(xì)胞計(jì)數(shù)（18.20±8.14）×10~6、嗜酸性粒細(xì)胞計(jì)數(shù)（3.80±1.75）×10~6明顯低于哮喘組大鼠肺泡灌洗液沉渣中細(xì)胞總數(shù)（10~8.92±27.55）×10~6、中性粒細(xì)胞計(jì)數(shù)（31.30±6.00）×10~6、嗜酸性粒細(xì)胞計(jì)數(shù)（7.10~±2.89）×10~6（P0.01）；對(duì)照組大鼠肺泡灌洗液沉渣中細(xì)胞總數(shù)（47.20±18.23）×10~6、中性粒細(xì)胞計(jì)數(shù)（7.40±1.96）×10~6、嗜酸性粒細(xì)胞計(jì)數(shù)（0.80±0.79）×10~6低于VD2組（P0.01）；VD2組肺組織ICAM-1表達(dá)（0.23±0.03）較哮喘組（0.34±0.10~）明顯降低（P0.01）；VD2組大鼠肺組織ICAM-1表達(dá)（0.23±0.03）高于對(duì)照組（0.17±0.38）（P0.01）。 結(jié)論：維生素D2能抑制Th2細(xì)胞因子IL-4水平，抑制哮喘大鼠氣道ICAM-1水平，減少肺內(nèi)氣道炎癥細(xì)胞浸潤(rùn)，減輕哮喘氣道炎癥反應(yīng)。
[Abstract]:Objective: asthma is a chronic airway inflammatory response disease involving a variety of cells, especially mast cells, eosinophil T lymphocytes, neutrophilic granulocytes, airway epithelial cells and various cellular components. Immune regulation plays an important role in the pathogenesis of asthma. Vitamin D plays a more and more important role in the immune regulation. It has been found that vitamin D can regulate the balance of Th1/Th2. In this experiment, vitamin D _ 2 was used to interfere with asthma rat model. The levels of IL-4 in alveolar lavage fluid, cell count and classification in alveolar lavage fluid, and the expression of ICAM-1 in lung were compared in each group. To investigate the anti-inflammatory effect of vitamin D in asthmatic rats and its possible mechanism. Methods: according to the principle of random grouping, 30 SD male rats were randomly divided into three groups: asthma group, vitamin D2 group and control group. Asthma group (n = 10) was sensitized by intraperitoneal injection of 10OVA1ml (containing ovalbumin 100mg and 100mg aluminum hydroxide) on the 15th day of the first day, and was stimulated by 1%OVA atomization inhalation in a closed atomizing tank on the 16th day, and continued to atomize for 1 week. After 30 minutes of arousing every day, they developed shortness of breath, wheezing or dyspnea) vitamin D2 VD2) group: the asthma group received intraperitoneal injection of vitamin D 25 脳 10 5 U / L 路kg 路dL from the 15th day, once every other day. The control group was treated with normal saline instead of OVA and vitamin D 2. The rats were killed within 24 hours after the last stimulation, and the lung tissue and alveolar lavage fluid were collected. Supernatant and precipitate cells were isolated from BALF after centrifugation, and lung tissue was treated with dehydration and translucent, paraffin embedded. The level of IL-4 in the supernatant of BALF was determined by Elisa method, and the precipitation cells of BALF were counted by modified Niu's counter and stained by smear. The level of intercellular adhesion molecule (ICAM 1) in rat lung tissue was determined by immunohistochemical method. The index data of each group were analyzed and compared by statistical method. Results the IL-4 content in the VD2 group was significantly lower than that in the asthmatic group (615.37 鹵94.77pgml / ml) and the IL-4 content in the VD2 group was significantly lower than that in the control group (72.80 鹵24.44) 脳 10 ~ (-6), the neutrophil count was 18.20 鹵8.14 脳 10 ~ (-6), and the eosinophil count was 3.80 鹵1.75 脳 10 ~ (-6). The expression of ICAM-1 in lung tissue of VD2 group was significantly lower than that of VD2 group (0.23 鹵0.03), which was significantly lower than that of asthmatic group (0.34 鹵0.10). The expression of ICAM-1 in lung tissue of VD2 group was significantly lower than that of control group (0.17 鹵0.38). Conclusion: vitamin D 2 can inhibit the level of Th2 cytokine IL-4, inhibit the level of airway ICAM-1 in asthmatic rats, reduce the infiltration of airway inflammatory cells in the lung, and alleviate the airway inflammation of asthma.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R725.6

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 馬雅心;戚好文;;維生素D3聯(lián)合地塞米松對(duì)哮喘大鼠的治療作用及機(jī)制[J];第四軍醫(yī)大學(xué)學(xué)報(bào);2008年15期

2 王曉芳;洪建國(guó);周小建;;維生素D對(duì)大鼠哮喘模型氣道炎癥的影響[J];上海醫(yī)學(xué);2008年01期

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