AQP1、VEGF在变应性鼻炎大鼠鼻黏膜中的表达及其相关性研究

发布时间：2018-08-10 19:41

【摘要】：研究目的水通道蛋白(Aquaporins, AQPs)是一类介导水、甘油、尿素等分子跨膜转运的疏水性跨膜蛋白,广泛分布于动、植物及微生物,迄今为止在哺乳动物体内已经发现了13个亚型,即AQP0-12。AQP1表达于多种上皮组织和内皮细胞。血管内皮生长因子(vascularendothelial growth factor,VEGF)是已知最强的血管通透因子,其通透作用强于组胺1×106倍,对血管生长具有强烈的诱导作用,可增生形成微血管,同时可作用于血管内皮细胞调节小血管和毛细血管的通透作用,这种作用使血浆及部分大分子物质(如纤维蛋白原)渗到血管外基质中致使组织发生水肿。研究发现,两者的异常表达和调节失控与某些疾病的炎性、水肿有关系,但在变应性鼻炎(allergic rhinitis, AR)中鲜有研究,且相关的变态反应机制尚不清楚。本实验旨在通过对照AQP1和AEGF在AR大鼠、正常大鼠以及地塞米松干预后AR大鼠鼻黏膜中的表达水平和两者的相关性,从而进一步探讨AQP1、AEGF在AR发病机制中的作用。研究方法选取8周龄健康Wistar大鼠30只,雌雄各半,体重160g-200g。正常饲养一周后随机分为三组,对照组,模型组和干预组,每组10只,以卵清蛋白(Ovalbumin,OVA)经典致敏法制作模型组和干预组Wistar大鼠AR模型,对照组以生理盐水代替OVA,干预组大鼠造模成功后腹腔注射地塞米松,干预一周,模型组以等量生理盐水代替。用组织化学方法观察变应性鼻炎的组织形态学特征,运用免疫组织化学方法(SP法)和real-time PCR两种方法分别测定各组鼻腔黏膜AQP1与VEGF的表达。实验结果以x±s表示,采用SPSS13.0统计分析软件对三组数据进行分析,数据符合正态分布且方差齐的进行方差分析及多组均数差别的多重比较(LSD法)。偏态或方差不齐的数据进行多组数据的Kruskal-Wallis检验及Bonferroni法两两比较平均秩和,AQP1与VEGF双变量符合正态分布,二者之间的相关性采用Pearson分析,反之采用Spearman秩相关。以P0.05为有统计学意义。研究结果按行为学评分标准,模型组和干预组大鼠均被激发出典型的症状,示造模成功。干预组经过地塞米松干预后,偶有流涕,喷嚏和挠鼻现象基本消失。造模成功后上皮杯状细胞明显增多,上皮下层组织水肿,炎性细胞浸润及嗜酸性细胞明显增多,腺体增生,小血管增加、充血。干预组组织水肿,血管扩张、腺体增生及炎性细胞浸润程度均减轻。三组大鼠的症状学分析模型组评分高于对照组和干预组。AQP1主要表达于大鼠鼻黏膜被覆上皮基底膜、血管内皮细胞膜和腺体细胞内,VEGF主要表达于小动、静脉血管内皮细胞和腺体细胞内。AQP1与VEGF在模型组中经免疫组织化学和real-time PCR法检测均高表达,干预组表达低于模型组。差异均具有统计学意义,且两者具有正相关性。结论AQP1、VEGF在实验性变应性鼻炎大鼠鼻粘膜的强表达,表明AQP1、VEGF参与了变应性鼻炎的发病过程,与组织的水肿、腺体的过度分泌可能有关。糖皮质激素可以明显下调AQP1、VEGF的表达水平,提示糖皮质激素可能是通过改变AQP1、VEGF的表达,减少腺体的分泌和改善鼻塞症状。且AQP1与VEGF之间具有正相关性,但具体机制尚不明确。
[Abstract]:Aquaporins (AQPs) are a class of hydrophobic transmembrane proteins that mediate the transmembrane transport of water, glycerol, urea and other molecules. AQPs are widely distributed in animals, plants and microorganisms. So far, 13 subtypes have been found in mammals, namely AQP 0-12. AQP 1 is expressed in many kinds of epithelial tissues and endothelial cells. Vascular endothelial growth factor (VEGF) is the strongest known vascular permeability factor. Its permeability is stronger than that of histamine by 1 65 It was found that the abnormal expression and regulation of fibrinogen were related to the inflammation and edema of some diseases, but little research was done in allergic rhinitis (AR), and the mechanism of allergic reaction was not clear. The expression of AQP1 and AEGF in the nasal mucosa of AR rats, normal rats and AR rats after dexamethasone intervention were compared, so as to further explore the role of AQP1 and AEGF in the pathogenesis of AR. Three groups, control group, model group and intervention group, 10 rats in each group, Ovalbumin (OVA) classical sensitization model group and intervention group Wistar rats AR model, control group with normal saline instead of OVA, intervention group rats after successful modeling intraperitoneal injection of dexamethasone, intervention for a week, model group with the same amount of normal saline instead. Methods The histomorphological features of allergic rhinitis were observed and the expressions of AQP1 and VEGF in nasal mucosa were detected by immunohistochemical method (SP) and real-time PCR respectively. Variance analysis and multiple comparisons of mean differences (LSD). Kruskal-Wallis test and Bonferroni method were used to compare the mean rank sums of skewed or uneven data. AQP1 and VEGF bivariates were in normal distribution. Pearson analysis was used to analyze the correlation between the two variables, and Spearman rank correlation was used instead. Results According to the behavioral scoring criteria, the rats in the model group and the intervention group were all stimulated with typical symptoms, indicating that the model was successful. The degree of edema, vasodilation, glandular hyperplasia and inflammatory cell infiltration were alleviated in the intervention group. The scores in the model group were higher than those in the control group and intervention group. AQP1 was mainly expressed in the basement membrane of nasal mucosa and vascular endothelial cells. AQP1 and VEGF were highly expressed in the model group by immunohistochemistry and real-time PCR. The expression of AQP1 and VEGF in the intervention group was lower than that in the model group. The strong expression of AQP1 and VEGF in nasal mucosa of rats with rhinitis suggests that AQP1 and VEGF are involved in the pathogenesis of allergic rhinitis and may be related to tissue edema and excessive secretion of glands. There is a positive correlation between AQP1 and VEGF, but the specific mechanism is not yet clear.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R765.21

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