本文關(guān)鍵詞：Toll樣受體2和Toll樣受體4在小鼠變應(yīng)性鼻炎中的表達(dá)　出處：《中國醫(yī)科大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： TLR2 TLR4 變應(yīng)性鼻炎 小鼠模型 鼻黏膜

【摘要】： 目的 變應(yīng)性鼻炎是一種吸入變應(yīng)原而導(dǎo)致的以鼻黏膜嗜酸性粒細(xì)胞、肥大細(xì)胞、T淋巴細(xì)胞等多種炎性細(xì)胞浸潤,IgE介導(dǎo)的免疫細(xì)胞釋放化學(xué)介質(zhì)引起的Ⅰ型變態(tài)反應(yīng)炎癥性疾病,以鼻癢、打噴嚏、流清涕等為主要癥狀,可并發(fā)哮喘、鼻-鼻竇炎和分泌性中耳炎等疾病,嚴(yán)重影響患者生活質(zhì)量。近年來對變應(yīng)性鼻炎的研究雖然有了很大進(jìn)展,但對其發(fā)病機(jī)制仍需要進(jìn)一步的研究。Toll樣受體(TLRs)是近年發(fā)現(xiàn)的介導(dǎo)天然免疫反應(yīng)的關(guān)鍵受體,它們識別特定的病原相關(guān)分子模式并啟動了一系列防御反應(yīng)。由于TLR2和TLR4的配體已經(jīng)涵蓋了自然界絕大多數(shù)對人體致病的微生物類別,因此,二者在人Toll樣受體家族中最為重要。以前的研究已經(jīng)證實TLR2和炎癥因子之間的協(xié)同促進(jìn)表達(dá)在炎癥中的重要意義。目前,對于TLRs在變應(yīng)性鼻炎中的表達(dá)及作用機(jī)制的研究報道少見,且存在爭議。國內(nèi)報道TLRs在鼻息肉中有高表達(dá),也有國外報道沒有發(fā)現(xiàn)TLRs在鼻息肉組較健康對照組有高表達(dá),但在變應(yīng)性鼻炎的急性期發(fā)現(xiàn)高表達(dá)。由此可見,對TLRs在變應(yīng)性鼻炎中表達(dá)的研究有重要意義。 方法 1、變應(yīng)性鼻炎模型制備 Bal b／c小鼠以卵清白蛋白(OVA)100μg加氫氧化鋁(Al(OH)3)2mg溶于0.1ml的生理鹽水中腹腔注射,分別在1、3、5、7、9、11、13、15天各一次,共8次,然后于第16天起每側(cè)鼻腔用10μg卵清白蛋白滴鼻激發(fā),連續(xù)11天,建立小鼠變應(yīng)性鼻炎模型。 2、變應(yīng)性鼻炎模型評價 (1)末次滴鼻激發(fā)后觀察30分鐘,觀察小鼠打噴嚏,撓鼻,抓臉,鼻溢等癥狀并進(jìn)行計分；(2)抽取致敏動物血,血清中加入標(biāo)記熒光生物素的變應(yīng)原,然后在450nm處測定光密度值,來判定血清中IgE的效價；(3)各組小鼠模型行常規(guī)HE染色,并行高倍鏡下嗜酸性粒細(xì)胞計數(shù)。 3、應(yīng)用免疫組織化學(xué)染色技術(shù)在蛋白水平對變應(yīng)性鼻炎的Toll樣受體2、4的表達(dá)進(jìn)行研究。 4、統(tǒng)計結(jié)果均以均數(shù)±標(biāo)準(zhǔn)差表示,多組均數(shù)間比較用OneWay-ANOVA分析。均用SPSS 13.0軟件進(jìn)行統(tǒng)計分析。 結(jié)果 1、成功的制備出小鼠變應(yīng)性鼻炎的模型。 2、變應(yīng)性鼻炎組小鼠的癥狀評分與正常對照組比較差異有統(tǒng)計學(xué)意義(P0.01)；變應(yīng)性鼻炎組小鼠IgE效價與正常對照組比較差異有統(tǒng)計學(xué)意義(P0.01)；變應(yīng)性鼻炎組小鼠的嗜酸性粒細(xì)胞計數(shù)與正常對照組比較差異有統(tǒng)計學(xué)意義(P0.01)。 3、免疫組化法發(fā)現(xiàn)TLR2和TLR4在正常小鼠鼻黏膜細(xì)胞的胞漿和胞核無陽性或極弱陽性染色；在變應(yīng)性鼻炎組小鼠鼻黏膜細(xì)胞胞核和／或胞漿呈不同程度的褐色或黃棕色陽性染色。 4、變應(yīng)性鼻炎組小鼠在30min時鼻黏膜細(xì)胞上TLR2表達(dá)開始增多,但與正常對照組差異無統(tǒng)計學(xué)意義(P0.05),2h時鼻黏膜細(xì)胞上TLR2表達(dá)明顯增加(P0.01),12h時鼻黏膜細(xì)胞上TLR2表達(dá)開始下降(P0.05)。24h時鼻黏膜細(xì)胞上TLR2表達(dá)與正常對照組差異無統(tǒng)計學(xué)意義(P0.05)。 5、變應(yīng)性鼻炎組小鼠在30min時鼻黏膜細(xì)胞上TLR4表達(dá)開始增多,與正常對照組差異有統(tǒng)計學(xué)意義(P0.05),2h時鼻黏膜細(xì)胞上TLR4表達(dá)明顯增加(P0.01),12h時鼻黏膜上TLR4表達(dá)開始下降(P0.05)。24h時鼻黏膜細(xì)胞上TLR4表達(dá)正常對照組差異無統(tǒng)計學(xué)意義(P0.05)。 結(jié)論 1、TLR2和TLR4在變應(yīng)性鼻炎組中均有表達(dá)。 2、TLR2和TLR4在變應(yīng)性鼻炎的急性期出現(xiàn)高表達(dá),在變應(yīng)性鼻炎的整個病程中呈現(xiàn)動態(tài)變化。
[Abstract]:Purpose Toll - like receptor ( TLRs ) is the key receptor in human Toll - like receptor family . Toll - like receptor ( TLRs ) is one of the most important receptors in human Toll - like receptor family . Toll - like receptor ( TLRs ) is a key receptor in human Toll - like receptor family . Toll - like receptor ( TLRs ) is a key receptor in human Toll - like receptor . method 1 . Preparation of allergic rhinitis model Balb / c mice were injected intraperitoneally with 100 渭g of aluminum hydroxide ( Al ( OH ) 3 ) 2 mg dissolved in 0.1 ml of physiological saline , once every 1 , 3 , 5 , 7 , 9 , 11 , 13 , 15 days , and then stimulated with 10 渭g of egg albumin nasal drops on each side of the nasal cavity on day 16 , and the allergic rhinitis model was established for 11 days . 2 . Evaluation of allergic rhinitis model ( 1 ) observing 30 minutes after the last nose - nose excitation , observing the symptoms and scoring of the mice , scratching , nose , face , rhinorrhea and the like ; ( 2 ) extracting the sensitized animal blood , adding the allergen of the labeled fluorescent biotin in the serum , and then measuring the optical density value at 450 nm to determine the titer of the IgE in the serum ; and ( 3 ) performing routine HE staining in each group of mouse models , and counting the eosinophils in the parallel high - magnification mirror . 3 . The expression of Toll - like receptor 2 , 4 in allergic rhinitis was studied by immunohistochemical staining . 4 . The statistical results were expressed by mean 鹵 standard deviation , and one Way - ANOVA was used to analyze the multiple groups . The statistical analysis was carried out with SPSS 13.0 software . Results 1 . The model of mouse allergic rhinitis was successfully prepared . 2 . There was significant difference between the symptom scores of allergic rhinitis group and the normal control group ( P0.01 ) , the IgE titer of allergic rhinitis group was significantly different from that of the normal control group ( P0.01 ) , and the count of eosinophils in allergic rhinitis group was significantly different from that of the normal control group ( P0.01 ) . 3 . Immunohistochemical method showed that TLR2 and TLR2 were not positive or weakly positive staining in the cytoplasm and nuclei of normal mouse nasal mucosa cells , and the nuclei and / or cytoplasm of nasal mucosa cells of allergic rhinitis group were stained with brown or yellowish brown positive . 4 . The expression of TLR2 in nasal mucosa increased significantly after 30 min in allergic rhinitis group ( P0.05 ) . The expression of TLR2 in nasal mucosa increased significantly ( P0.05 ) at 2h . The expression of TLR2 in nasal mucosa was not statistically significant at 24h ( P0.05 ) . 5 . In allergic rhinitis group , the expression began to increase in nasal mucosa cells at 30 min . There was a significant difference between nasal mucosa and normal control group ( P0.05 ) , and the expression began to decrease in nasal mucosa at 12 h ( P0.05 ) . Conclusion 1 . TLR2 and TLR2 were expressed in allergic rhinitis group . 2 . TLR2 and TLR2 were highly expressed in the acute stage of allergic rhinitis and showed dynamic changes in the whole course of allergic rhinitis .

【學(xué)位授予單位】：中國醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R765.21

【參考文獻(xiàn)】

相關(guān)期刊論文 前8條

1 孫守勛;蒲曉允;;Toll樣受體2的研究進(jìn)展[J];重慶醫(yī)學(xué);2008年05期

2 王成碩,董震,楊占泉;Toll樣受體mRNA在鼻息肉中的表達(dá)及意義[J];中國耳鼻咽喉頭頸外科;2004年03期

3 錢彥方,李炳文,周正謀,王嘉玲,李進(jìn)讓,王鳳梅,周蘭坤;克鼻敏對豚鼠超敏鼻炎模型電鏡光鏡的病理觀察[J];北京中醫(yī)藥大學(xué)學(xué)報;1999年06期

4 殷明德;;變應(yīng)性鼻炎對耳鼻咽喉科的影響[J];臨床耳鼻咽喉頭頸外科雜志;2007年17期

5 余洪猛,文三立,劉志剛,方勇;蛔蟲變應(yīng)原致過敏性鼻炎豚鼠模型建立[J];上海實驗動物科學(xué);2000年04期

6 余洪猛,文三立,劉志剛,方勇,王華;豚草花粉變應(yīng)原致過敏性鼻炎豚鼠模型的建立[J];上海實驗動物科學(xué);2001年04期

7 王成碩,董震;Toll樣受體mRNA在慢性鼻竇炎患者鼻黏膜上皮細(xì)胞中的表達(dá)[J];中華耳鼻咽喉科雜志;2003年04期

8 徐睿,李源,謝民強(qiáng),許庚,張革化,王樹芳;白介素-5(IL-5)在人類鼻息肉組織中的含量、分布和表達(dá)(英文)[J];Chinese Medical Journal;2002年09期

，

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