【作者】 张秀娟；

【导师】 吴玉瑛；

【作者基本信息】 郑州大学 ， 耳鼻咽喉科学， 2010， 硕士

【摘要】 背景和目的鼻息肉(nasal polyps, NP)是鼻腔和鼻窦粘膜的慢性炎症性疾病,发病率约占总人数的1%～4%。临床上以慢性鼻-鼻窦炎伴鼻息肉(CRSwNP)较为多见,其复发率较高。发病机制目前尚不清楚,近年来的研究表明可能与感染、变态反应、免疫及细胞因子等有关。叉头状转录因子3(forkhead transcription factor p3, Foxp3)是一个新发现的转录调节因子,对CD4+ CD25+ T调节性细胞的发育及功能的维持至关重要,并具有免疫抑制作用。FOXP3已被证明在CD4+ CD25+ T调节性细胞上特异性表达,在一定程度上反映CD4+ CD25+ T调节性细胞的水平和功能活性。本研究旨在分析FOXP3、CD4和CD8在鼻息肉中的表达及分布、弄清FOXP3、与CD4和CD8的相互关系,探讨FOXP3+ CD4+ CD25+ T调节细胞、T淋巴细胞亚群在鼻息肉发生、发展中的作用,明确其临床意义。材料与方法本研究对实验组50例鼻息肉组织和对照组20例正常下鼻甲黏膜组织的标本切片分别行HE染色,采用免疫组化SP法检测两组标本中FOXP3、CD4以及CD8的表达情况,采用SPSS16.0统计软件,选择正确的检验方法,进行数据分析及对照研究。结果1 FOXP3蛋白的阳性产物主要定位于细胞核和(或)细胞质,呈棕黄色或棕褐色颗粒。免疫组化结果光镜下见：鼻息肉组和正常下鼻甲黏膜组均能观察到FOXP3阳性细胞。2 CD4和CD8蛋白阳性产物主要定位于细胞膜,呈棕黄色或棕褐色颗粒。在鼻息肉组织中见大量的CD4和CD8阳性表达的细胞,呈丛集性分布,多集中在上皮下和腺体周围,CD4+T细胞数显著高于CD8+T细胞数。正常下鼻甲黏膜组织中大部分切片也可见少量的CD4+T细胞和CD8+T细胞。3计算机图像分析系统检测FOXP3、CD4、CD8平均灰度值(1)FOXP3在50例鼻息肉组织和20例正常下鼻甲黏膜组织中表达的平均灰度值差异具有统计学意义(p<0.05)。在单侧和双侧鼻息肉组织中FOXP3表达的平均灰度值差异具有统计学意义(p<0.05)。FOXP3在鼻息肉首发组与复发组表达的平均灰度值差异具有统计学意义(p<0.05)。(2)CD4在50例鼻息肉组织和20例正常下鼻甲黏膜组织中表达的平均灰度值差异具有统计学意义(p<0.01)。在单侧与双侧鼻息肉组织中CD4表达的平均灰度值差异具有统计学意义(p<0.05)。CD4在鼻息肉首发组与复发组表达的平均灰度值差异具有统计学意义(p<0.05)。(3)CD8在50例鼻息肉组织和20例正常下鼻甲粘膜组织中表达的平均灰度值差异具有统计学意义(p<0.05)。在单侧与双侧鼻息肉组织中CD8表达的平均灰度值差异具有统计学意义(p<0.05)。CD8在鼻息肉首发组与复发组表达的平均灰度值差异具有统计学意义(p<0.05)。4通过spearson相关分析,在鼻息肉组织中CD4与FOXP3之间存在负相关关系(γ=-0.383,p<0.01)；CD8与FOXP3之间存在负相关关系(r=-0.364,p<0.01)。结论1FOXP3在鼻息肉组织中表达下调,提示FOXP3的缺乏或功能低下在鼻息肉的发生和发展中起着重要的作用。2FOXP3、CD4和CD8均参与鼻息肉的形成过程,且可能鼻息肉的复发特性有关。3FOXP3、CD4和CD8在鼻息肉中的表达具有相关性,提示三种蛋白可能协同参与鼻息肉的形成过程。4检测鼻息肉组织中FOXP3表达,可以了解鼻息肉微环境中CD4+ CD25+调节性T细胞浸润状态,从而可判定鼻息肉组织局部免疫反应状态,为鼻息肉提供新的治疗方法。更多还原

【Abstract】 Background and ObjectiveNasal polyps is a kind of chronic inflammatory disease of nasal cavity and nasal sinuses mucosa, the total number of its incidence is about 1%-4%. Nasal polyps usually occur in the lateral wall of nasal cavity and nasal roof, Second, ethmoid and membranous of maxillary sinuses are also seen, and the rate of its recurrence was high. The etiology and pathogenesis of nasal polyps are not yet clear, recent research indicates that it may be related to infection, allergy, immunology and cell factor and so on.Forkhead transcription factor p3 is a newly discovered transcription factor, its a multifunctional transcription factor which is related to the growth and development of CD4+ CD25+ Treg cells. FOXP3 has been shown specifically expressed in CD4+ CD25+ T regulatory cells, to some extent, it reflects the level and functional activity of the CD4+ CD25+ T regulatory cells.The purpose of this study is to analysis the expression and distribution of FOXP3、CD4、CD25+ Treg cells, to explore the effects in occurrence,development of nasal polyps, to determine its clinical significance.Methods50 cases of nasal polyps(the test group) and 20 cases of normal inferior turbinate mucosa (the normal group) underwent HE staining and immunohistochemical (SP method detected) in two groups to detect the expression of FOXP3, CD4 and CD8, using statistical packages of SPSS16.0,the datas will be studied by classification, analysis and contrast.Results1 The positive products of FOXP3 protein localized in the cytoplasm and (or)cytoplasm, and the color was clearly brownish yellow or brown particles. Under the microscope, the FOXP3 positive cell could be observed in both of nasal polyps and normal turbinate mucasa specimemts.2 The positive staining of CD4 protein and CD8 protein mainly located in the cell membrane, and were stained for brown yellow or brown particles. In nasal polyps, a large number of CD4 and CD8 positive lymphocytes can be seen, wich gathered in the skin and around the gland, and was scattered distribution in the interstitium. CD4+Tcells was significantly higher than the number of CD8+Tcells. In normal mucosa group, most of the slices can also be found, but the quantity of CD4+T cells and CD8+T cells is less which concentrated in the glands around the blood vessels, and scattered.3 Computer graphics analysis system detects FOXP3, CD4, CD8 average gray value(1) The difference of average gray value of FOXP3 between 50 cases of nasal polyps and 20 cases of inferior turbinate mucosa was statistically significant (p< 0.05). The difference of average gray value of FOXP3 between unilateral and bilateral nasal polyps was statistically significant(p<0.05). The difference of average gray value of FOXP3 between starting group and relapse group wa statistically significant (p<0.05).(2) The difference of average gray value of CD4 between 50 cases of nasal polyps and 20 cases of inferior turbinate mucosa was statistically significant (p< 0.05). The difference of average gray value of CD4 between unilateral and bilateral nasal polyps was statistically significant (p<0.05). The difference of average gray value of CD4 between starting group and relapse group wa statistically significant (p <0.05).(3) The difference of average gray value of CD8 between 50 cases of nasal polyps and 20 cases of inferior turbinate mucosa was statistically significant (p< 0.05). The difference of average gray value of CD8 between unilateral and bilateral nasal polyps was statistically significant (p<0.05). The difference of average gray value of CD8 between starting group and relapse group wa statistically significant (p <0.05). 4. By spearson correlation analysis, there exists the negative correlation between FOXP3 and CD4 in nasal polyps(r=-0.383,P=0.006); nor exists between CD8 and FOXP3 (r=-0.364,p=0.009).Conclusion1 The expression of FOXP3 in nasal polyps reduced, suggesting that the absence or dysfunction of FOXP3 plays an important role in the occurrence and development of nasal polyps.2 The FOXP3, CD4 and CD8 were involved in the formation of nasal polyps, and maybe related to the recurrence properties of nasal polyps.3 The expression of FOXP3 CD4 and CD8 in nasal polyps correlated to each other, indicating that the three proteins might collaboratively involved in the formation of nasal polyps.4 By detecting the expression of FOXP3 in nasal polyps, we can know the state of CD4+ CD25+ regulatory T cell infiltration in nasal microenvironment, and determine the state of local immune response in nasal polyps, which provide a new method of treatment for nasal polyps.更多还原