【作者】 刘建国；

【导师】 刘月辉；

【作者基本信息】 南昌大学 ， 耳鼻咽喉—头颈外科， 2008， 硕士

【摘要】 目的:建立长时间抗原攻击SD大鼠变应性鼻炎(allergic rhinitis,AR)动物模型研究抗原攻击后鼻黏膜组织病理形态改变特点。方法:180只健康SD大鼠雌雄不分每组10只随机分正常对照组和AR组,AR组按实验设计又分为:间歇致敏组和持续激发组。AR模型造模方法:首先在SD大鼠的腹腔注入卵清蛋白和氢氧化铝的混合液进行基础致敏,然后再向鼻腔滴入5%的卵清蛋白溶液进行加强激发。分别于1、2、4、8、12、16周处死大鼠10只,取材。用苏木精-尹红(HE)染色、血管铸形方法观察AR鼻粘膜中组织学变化;用透射、扫描电镜观察AR鼻粘膜超微结构的变化;用免疫组织化学(SABC)、逆转录聚合酶联反应(RT-PCR)方法检测相关因子在两实验组和对照组大鼠鼻粘膜中的表达特点。结果:与正常对照组相比AR组大鼠随着时间的延长鼻黏膜组织出现不同病理形态学的特点:(1)抗原激发加强后早期(1至4周):间歇激发组出现上皮细胞水肿、脱落,粘膜层组织间水肿明显,杯状细胞化生、肥大,基底层胶原水肿明显,固有层血管明显扩张,腺体增生、肥大、分泌旺盛,粘膜层增高和结构明显紊乱;持续激发组上述组织病理形态改变进展更明显,尤其是上皮细胞的剥脱、黏膜层增高和结构紊乱,但是早期EOS的浸润数量、HO-1mRNA的表达量和TGF-β1、FGF-2蛋白表达量持续组较间歇组高但两组间比较没有显著的差别(P>0.05)。(2)在抗原激发加强后期(8至16周):间歇激发组出现再生修复:上皮细胞、组织间水肿出现明显好转,纤毛排列恢复整齐,粘膜层细胞排列趋向整齐,肥大的杯状细胞明显减少,腺体体积和扩张的腺管趋于正常,固有层的炎症细胞浸润减轻但是不能完全消除;持续激发组鼻粘膜上皮层损伤有一定程度修复,但是修复后的上皮排列不整齐,杯状细胞体积增大,粘膜下层血管增多,血管壁增厚。腺体体积增大,腺管结构模糊,黏膜有大量的淋巴细胞浸润,在粘膜层尤其是基底膜层出现大量胶原沉积和组织中纤维成分增多。EOS浸润数和HO-1mRNA表达量和TGF-β1、FGF-2蛋白表达量持续组与间歇、正常对照组比较出现了显著的增高(P<0.05)。另外,在该阶段持续激发组随着黏膜病变的过程,出现症状减轻的趋势,即症状积分明显下降,而间歇组下降更加明显。结论:本试验AR大鼠鼻黏膜中间歇组和持续组在致敏加强后早期鼻粘膜均表现为炎性损伤为主,且都呈进行性,但持续组更明显;后期持续组鼻粘膜组织病理形态改变表现为一种慢性炎症过程,在间歇激发组鼻黏膜组织病理变化恢复接近正常,主要表现为可复性修复,这可能与间歇组大鼠脱离变应原后可以出现鼻粘膜结构和功能的自行恢复;而持续组持续受抗原攻击主要表现为粘膜组织增生和瘢痕修复等组织重构特点:上皮细胞持续剥脱,上皮下纤维化,杯状细胞增生,血管增生、管壁不规则增厚、腺体体积增大和增生,基底膜层胶原沉积增多;TGF-β1、FGF-2都参与了AR鼻黏膜组织修复;随着鼻粘膜变态性炎症后组织重构,鼻部症状有减轻的趋势。另外,与正常组比较,间歇组和持续组大鼠鼻粘膜的炎症和氧化损伤早期呈现加重,后期有减轻的趋势,持续组没有间歇组明显。更多还原

【Abstract】 [Objective] To establish the model of allergic rhinitis(AR) of Sprague-Dawley(SD) rats and observe the damage of nasal mucosa in morphology and the characteric of pathologic change in nasal mucosa of AR .[Methods] Total 180 healthy SD rats irrespective of female and male were randomly devided into the normal and AR groups it including intermittent allergic group and persistence group respectively . After establishing the model of allergic rhinitis by intraperitoneal injecting comphensive solution of ovalbumin(OVA)and AL(OH)3 then drope 5%OVA solution to nose. Sacrifice rats after 1、2、4、8、12、16weeks ,Apply hematoxylin- ereuth dyeing、make microvascular corrosion casts、transmission electrical microscope to analysis ultrastructure and apply immunohistochemistry technique(SABC method)、reverse transcriptio polymerase chain reaction technique to study the protein expressiong of HO-1、TGF-β1、FGF-2 in nasal mucosa AR.[Results] Compare to normal rats,intermittence group: 1)early 4 weeks after intensively sensitizating characteric of morph: endothelial cell edemata、desquamate,textus edemata in lamina epithelialis, goblet cells metaplasia、hypertrophy、productive secretion, angiectasis greatly in propria lamina, glandular organ hypertrophy, collagen oedemata in basal membrane, mucous layer thickening, chaotic textus strcture. Pseudostraified columnar ciliated epithelium or basal layer, with cilia loss and increasing inflammatory cell. eosinophils infiltrating and the expression of HO-1、TGF-β1、FGF-2 are difference significantly. 2) late8,12,16weeks:tissue edema significantly improved, mucosal cells tend to tidy and neatly arranged cilia, the hypertrophic goblet cells decreased significantly, the glands and expansive glandular tube tend to normal. infiltration of inflammatory cells in inherent layer reducing but not completely eliminated, fiber components increased at the same time. eosinophils infiltrating and the expression of HO-1、TGF-β1、 FGF-2 are gradually decreasing. Persistence group:1)early 4 weeks after intensively sensitizating characteric of morph : as well as intermittent group, mucosal injuried is very severe. but eosinophils infiltrating and the expres -sion of HO-1 are also not difference significantly compare to intermittence group(P>0.05)。2) late8,12,16 week:damaged epithelium repaired in a certain extent with fibration, but the restoration result of epithelial are untidiness, goblet cell volume is increased submucosal blood vessels increased, vascular wall thickening.especially in the mucous layer a large number of collagen deposited in basement membrane, gland volume increased、hyperplasia,glandular tube structure are fuzzy. further,EOS infiltrating and the expression of HO-1mRNA、TGF-β1、FGF-2 protein were significantly different compared to intermittence group(P <0.05). In the progress of remodeling the symptom of AR rats have the trend to reduce.[Conclusion] two groups of AR mucosa before 4 weeks there are mucosa damage,while after 4 weeks they are very different in pathologic change. compare to the intermittence group,the persistence group mucosa are remodeling to repair the damage of mucosa,it maybe the response of damage. Pathologiccharacteristic:repair after cell loss, subepithelial fibrosis, goblet cell proliferation, vascular proliferation, irregular thickening of the lumen, the gland volume increased, collagen deposition increasing in the basement membrane layer. While intermittence group rat mucosa remodeling aren’t significant, it maybe the recovery become more easy after deprivating allergen. In the progress of remodeling the nasal mucosa inflammation and oxidative damage have the trend to reduce .and TGF-β1、FGF-2 are participate in fibrosis repairing.更多还原