MMP-7、TGF-β1及PAI-1在鼻息肉中的表達(dá)及其病理意義
本文關(guān)鍵詞: 免疫組化 鼻息肉 MMP-7 TGF-β1 PAI-1 出處:《鄭州大學(xué)》2010年碩士論文 論文類型:學(xué)位論文
【摘要】: 鼻息肉是最常見的上呼吸道慢性炎癥性疾病之一,其病因及發(fā)病機(jī)制被大多數(shù)學(xué)者認(rèn)為是多因素多步驟的過程,與感染、免疫及細(xì)胞因子有關(guān)。鼻息肉的具體發(fā)病機(jī)制尚不完全清楚,但呼吸道黏膜的損傷-修復(fù)和結(jié)構(gòu)重塑已經(jīng)被證實(shí)在鼻息肉的發(fā)病機(jī)制中起關(guān)鍵作用,而細(xì)胞外基質(zhì)的合成和降解是組織結(jié)構(gòu)重塑中最關(guān)鍵的一步。為此我們檢測與組織結(jié)構(gòu)重塑及基底膜合成和降解過程密切相關(guān)的基質(zhì)金屬蛋白酶-7、轉(zhuǎn)化生子因子-β1及纖溶酶原激活物抑制劑-1在鼻息肉及正常下甲黏膜中的表達(dá)水平,以探討其在鼻息肉形成過程中發(fā)揮的作用。 材料和方法 1.臨床資料: 鼻息肉標(biāo)本50例,均為2008年7月-2009年7月在鄭州大學(xué)一附院接受鼻內(nèi)鏡鼻竇手術(shù)的患者。選擇標(biāo)準(zhǔn):單側(cè)或者雙側(cè)鼻竇CT檢查陽性,鼻內(nèi)鏡檢查或手術(shù)中發(fā)現(xiàn)息肉。所有患者臨床病例資料完整,均無變應(yīng)性鼻炎,支氣管哮喘,阿司匹林耐受不良及其他系統(tǒng)性疾病,且在術(shù)前一月內(nèi)未使用過激素或抗組胺類藥物。下鼻甲粘膜標(biāo)本30例,均為2008年7月-2009年7月在鄭州大學(xué)一附院接受鼻內(nèi)鏡手術(shù)的鼻中隔偏曲患者,取之對側(cè)代償性肥大的下甲。 2.實(shí)驗(yàn)方法: 對50例鼻息肉組織和30例下鼻甲粘膜組織進(jìn)行HE染色,并用免疫組化SP法檢測上述標(biāo)本中MMP-7、TGF-β1及PAI-1蛋白的表達(dá)。 3.結(jié)果判定: 三種蛋白質(zhì)均以胞質(zhì)呈現(xiàn)棕黃色或者棕褐色染色為陽性細(xì)胞。取染色良好(特異性染色清晰,背景染色低)的組織切片,根據(jù)染色細(xì)胞不同的染色強(qiáng)度和高倍鏡下陽性細(xì)胞數(shù)百分比作為依據(jù),其兩者相乘,結(jié)果大于3分極為免疫反應(yīng)陽性,進(jìn)行結(jié)果的判定。 4.統(tǒng)計(jì)學(xué)分析: 應(yīng)用SPSS11.0統(tǒng)計(jì)軟件行卡方檢驗(yàn)(x2檢驗(yàn))進(jìn)行顯著性檢驗(yàn),相關(guān)分析應(yīng)用列聯(lián)表的關(guān)聯(lián)性分析,檢驗(yàn)標(biāo)準(zhǔn)α=0.05。 結(jié)果 1.HE染色光鏡下見細(xì)胞核染色為藍(lán)色、藍(lán)黑色或者深藍(lán)色,而胞漿呈現(xiàn)為淡粉色或者是淡紅色。可見鼻息肉組織由極度水腫的疏松結(jié)締組織構(gòu)成,表面覆蓋呼吸上皮,大量杯狀細(xì)胞增生,其表面沒有纖毛,腺體組織增生明顯,多為長管狀或囊性擴(kuò)張,沒有真正的漿液黏液腺,間質(zhì)中有大量炎性細(xì)胞浸潤,主要為嗜酸性粒細(xì)胞,肥大細(xì)胞的數(shù)量也明顯增加,基底膜增厚,血管通透性增高,沒有神經(jīng)組織。而在下鼻甲黏膜組織中見到的腺管多呈圓形,間質(zhì)中炎癥細(xì)胞浸潤較少,幾乎見不到嗜酸性粒細(xì)胞。 2.MMP-7的陽性產(chǎn)物主要定位于細(xì)胞漿中,呈棕黃或者棕褐色,光鏡下可見其廣泛表達(dá)在鼻息肉組織中的上皮細(xì)胞、血管內(nèi)皮細(xì)胞、腺體細(xì)胞以及間質(zhì)內(nèi)浸潤的炎癥細(xì)胞的胞漿內(nèi),在基質(zhì)中也有表達(dá)。MMP-7其在鼻息肉中的陽性表達(dá)率為42.00%。而在對照組正常下甲粘膜中的陽性率為3.33%。兩者表達(dá)結(jié)果經(jīng)統(tǒng)計(jì)學(xué)分析顯示,p值少于0.05,說明其在鼻息肉及下甲組織中表達(dá)的差異具有統(tǒng)計(jì)學(xué)意義。 3.TGF-β1的陽性產(chǎn)物主要定位于細(xì)胞漿中,呈棕黃或者棕褐色,光鏡下可見其廣泛表達(dá)在鼻息肉組織中的上皮細(xì)胞、血管內(nèi)皮細(xì)胞、腺體細(xì)胞以及間質(zhì)內(nèi)浸潤的炎癥細(xì)胞的胞漿內(nèi),在基質(zhì)中也有表達(dá)。其在鼻息肉中的陽性表達(dá)率為72.00%。TGF-β1在對照組下鼻甲粘膜組織中的陽性率為13.33%。兩者表達(dá)結(jié)果經(jīng)統(tǒng)計(jì)學(xué)分析顯示,p值少于0.05,說明其在鼻息肉及下甲組織中的表達(dá)的差異具有統(tǒng)計(jì)學(xué)意義 4.PAI-1的陽性產(chǎn)物主要定位于細(xì)胞漿中,呈棕黃或者棕褐色,光鏡下可見其廣泛表達(dá)在鼻息肉組織中的上皮細(xì)胞、血管內(nèi)皮細(xì)胞、腺體細(xì)胞以及間質(zhì)內(nèi)浸潤的炎癥細(xì)胞的胞漿內(nèi),在基質(zhì)中也有表達(dá)。其在鼻息肉及對照組中的陽性表達(dá)率分別為18.00%和0.00%。兩者表達(dá)結(jié)果經(jīng)統(tǒng)計(jì)學(xué)分析顯示,p值少于0.05,說明其在鼻息肉及下甲組織中的表達(dá)的差異具有統(tǒng)計(jì)學(xué)意義。 5.MMP-7蛋白表達(dá)陽性病例中,TGF-β1蛋白表達(dá)陽性率為90.00%,MMP-7蛋白表達(dá)陰性病例中TGF-β1蛋白表達(dá)陽性率60.00%,經(jīng)統(tǒng)計(jì)學(xué)分析,結(jié)果顯示列聯(lián)系數(shù)r=0.311,p=0.0210.05,兩種蛋白在鼻息肉組織中的表達(dá)有關(guān)聯(lián)性。 6.MMP-7蛋白表達(dá)陽性病例中,PAI-1蛋白表達(dá)陽性率為40.00%,MMP-7蛋白表達(dá)陰性病例中PAI-1蛋白表達(dá)陽性率20.00%,經(jīng)統(tǒng)計(jì)學(xué)分析,結(jié)果顯示p0.05,兩種蛋白在鼻息肉組織中的表達(dá)無關(guān)聯(lián)性。 7.TGF-β1蛋白表達(dá)陽性病例中,PAI-1蛋白表達(dá)陽性率為30.56%,TGF-β1蛋白表達(dá)陰性病例中PAI-1蛋白表達(dá)陽性率0.00%,經(jīng)統(tǒng)計(jì)學(xué)分析,結(jié)果顯示列聯(lián)系數(shù)r=0.314,p0.05,兩種蛋白在鼻息肉組織中的表達(dá)有關(guān)聯(lián)性。 結(jié)論 1.異常增高的MMP-7,TGF-β1及PAI-1可使組織基底膜發(fā)生變化,參與鼻息肉的組織結(jié)構(gòu)重塑。 2.MMP-7、TGF-β1及PAI-1參與了鼻息肉的形成過程,但是與其臨床分型、患者的性別和年齡無關(guān)。
[Abstract]:Nasal polyps is one of the most common upper respiratory tract of chronic inflammatory disease, its etiology and pathogenesis by most scholars is a process of multiple factors and steps, and the infection, immunity and cytokines. The pathogenesis of nasal polyps is not completely understood, damage repair and remodeling of airway mucous membrane but has been that play a key role in the pathogenesis of nasal polyps, and the synthesis and degradation of extracellular matrix is the most critical step in tissue remodeling. Therefore we detected and tissue remodeling and basement membrane synthesis and degradation of matrix metalloproteinase is closely related to the -7, transforming growth factor beta 1 and plasminogen activator inhibitor -1 in nasal polyps and normal mucosa in a expression level, to explore the formation process play a role in nasal polyps.
Materials and methods
1. clinical data:
50 cases of nasal polyp specimens were in July 2008 -2009 year in July received endoscopic sinus surgery in the First Affiliated Hospital of Zhengzhou University. The patient selection criteria: unilateral or bilateral nasal polyps found CT positive, nasal endoscopy or surgery. All cases of patients with complete data, there were no allergic rhinitis, bronchial asthma, aspirin intolerance and other systemic diseases, and in the preoperative January in not using hormones or antihistamine drugs. 30 cases of inferior turbinate mucosa specimens were in July 2008 -2009 year in July received endoscopic sinus surgery in the First Affiliated Hospital of Zhengzhou University, the deviation of nasal septum were taken from contralateral compensatory hypertrophy under armour.
2. the method of experiment:
50 cases of nasal polyps and 30 cases of inferior turbinate mucosa were stained with HE, and the expression of MMP-7, TGF- beta 1 and PAI-1 protein in the above specimens were detected by immunohistochemical SP method.
3. the results are as follows:
Three proteins in the cytoplasm showed brown or brown staining in the cell. Good staining (specific staining clear, low background staining) tissue sections, according to the staining intensity of stained cells at high magnification and different number of positive cells percentage as the basis, the two multiplied by more than 3 points for immune results the positive reaction, to determine the results.
4. statistical analysis:
The SPSS11.0 statistical software was used for the saliency test (x2 test), the correlation analysis was applied to the correlation analysis of the contingency table, and the standard alpha =0.05. was tested.
Result
1.HE staining under light microscope nuclear staining is blue, dark blue or dark blue, and the cytoplasm is pink or reddish. Visible nasal polyps composed of loose connective tissue extreme edema, covering the surface of the respiratory epithelium, a large number of goblet cell hyperplasia, there is no cilia on the surface of glandular tissue, hyperplasia, mostly long or cystic dilatation, no real serous mucinous, interstitial inflammatory cell infiltration, mainly for eosinophils, mast cells also increased, basement membrane thickening, increased vascular permeability, no nerve tissue. And see in the inferior turbinate mucosa of the glandular tube round and less infiltration of inflammatory cells in the stroma, almost no eosinophils.
The positive products of 2.MMP-7 mainly located in the cytoplasm, showed brown or tan, under light microscope, which are widely expressed in nasal polyps epithelial cells, vascular endothelial cells, glandular cells and inflammatory cells infiltration in interstitial cytoplasm, in the matrix also has the expression of.MMP-7 positive expression in the nose polyps in the rate of 42.00%. in the positive rate of the control group under a normal mucosa. The results by statistical analysis showed the expression of 3.33%., P value of less than 0.05, indicating the significant difference in its expression in nasal polyps and a tissue.
Positive products of 3.TGF- beta 1 is mainly located in the cytoplasm, showed brown or tan, under light microscope, which are widely expressed in nasal polyps epithelial cells, vascular endothelial cells, glandular cells and inflammatory cells infiltration in interstitial cytoplasm, in the matrix also has expressed its positive expression. In nasal polyps was the positive rate of 72.00%.TGF- beta 1 in the control group under the turbinate mucosa tissues for the expression of 13.33%. results by statistical analysis showed that the p value is less than 0.05, indicating the significant difference in its expression in nasal polyps and under armour in the organization
The positive products of 4.PAI-1 mainly located in the cytoplasm, showed brown or tan, under light microscope, which are widely expressed in nasal polyps epithelial cells, vascular endothelial cells, glandular cells and inflammatory cells infiltration in interstitial cytoplasm, in the matrix are expressed. The positive expression in the nose the polyps in the control group were statistical analysis results showed that the expression of both 18% and 0.00%., P value of less than 0.05, indicating the significant difference in its expression in nasal polyps and Xiajia tissues.
The positive expression of 5.MMP-7 protein beta 1 cases, TGF- positive expression rate was 90%, negative cases of TGF- beta 1 protein expression rate of MMP-7 protein expression in 60%, by statistical analysis, results showed that the column connection number r=0.311, p=0.0210.05, two kinds of protein in nasal polyps of relevance.
The positive rate of PAI-1 protein expression was 40% in 6.MMP-7 protein positive cases. The positive rate of PAI-1 protein expression was 20% in negative cases of MMP-7 protein expression. After statistical analysis, the results showed that there was no correlation between the expression of P0.05 and two proteins in nasal polyps.
The positive expression of 7.TGF- protein beta 1 cases, the positive rate of PAI-1 expression was 30.56%, 1 protein expression of TGF- beta PAI-1 protein expression positive rate of 0% negative cases, through statistical analysis, results showed that the column connection number r=0.314, P0.05, the expression of two proteins in nasal polyps have relevance.
conclusion
1. the abnormal increase of MMP-7, TGF- beta 1 and PAI-1 can change the tissue basement membrane and participate in the remodeling of the tissue structure of nasal polyps.
2.MMP-7, TGF- beta 1 and PAI-1 are involved in the formation of nasal polyps, but their sex and age are not related to their clinical typing.
【學(xué)位授予單位】:鄭州大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2010
【分類號】:R765.25
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