【摘要】：研究目的水通道蛋白(Aquaporins, AQPs)是一類介導水、甘油、尿素等分子跨膜轉運的疏水性跨膜蛋白,廣泛分布于動、植物及微生物,迄今為止在哺乳動物體內已經發(fā)現了13個亞型,即AQP0-12。AQP1表達于多種上皮組織和內皮細胞。血管內皮生長因子(vascularendothelial growth factor,VEGF)是已知最強的血管通透因子,其通透作用強于組胺1×106倍,對血管生長具有強烈的誘導作用,可增生形成微血管,同時可作用于血管內皮細胞調節(jié)小血管和毛細血管的通透作用,這種作用使血漿及部分大分子物質(如纖維蛋白原)滲到血管外基質中致使組織發(fā)生水腫。研究發(fā)現,兩者的異常表達和調節(jié)失控與某些疾病的炎性、水腫有關系,但在變應性鼻炎(allergic rhinitis, AR)中鮮有研究,且相關的變態(tài)反應機制尚不清楚。本實驗旨在通過對照AQP1和AEGF在AR大鼠、正常大鼠以及地塞米松干預后AR大鼠鼻黏膜中的表達水平和兩者的相關性,從而進一步探討AQP1、AEGF在AR發(fā)病機制中的作用。研究方法選取8周齡健康Wistar大鼠30只,雌雄各半,體重160g-200g。正常飼養(yǎng)一周后隨機分為三組,對照組,模型組和干預組,每組10只,以卵清蛋白(Ovalbumin,OVA)經典致敏法制作模型組和干預組Wistar大鼠AR模型,對照組以生理鹽水代替OVA,干預組大鼠造模成功后腹腔注射地塞米松,干預一周,模型組以等量生理鹽水代替。用組織化學方法觀察變應性鼻炎的組織形態(tài)學特征,運用免疫組織化學方法(SP法)和real-time PCR兩種方法分別測定各組鼻腔黏膜AQP1與VEGF的表達。實驗結果以x±s表示,采用SPSS13.0統(tǒng)計分析軟件對三組數據進行分析,數據符合正態(tài)分布且方差齊的進行方差分析及多組均數差別的多重比較(LSD法)。偏態(tài)或方差不齊的數據進行多組數據的Kruskal-Wallis檢驗及Bonferroni法兩兩比較平均秩和,AQP1與VEGF雙變量符合正態(tài)分布,二者之間的相關性采用Pearson分析,反之采用Spearman秩相關。以P0.05為有統(tǒng)計學意義。研究結果按行為學評分標準,模型組和干預組大鼠均被激發(fā)出典型的癥狀,示造模成功。干預組經過地塞米松干預后,偶有流涕,噴嚏和撓鼻現象基本消失。造模成功后上皮杯狀細胞明顯增多,上皮下層組織水腫,炎性細胞浸潤及嗜酸性細胞明顯增多,腺體增生,小血管增加、充血。干預組組織水腫,血管擴張、腺體增生及炎性細胞浸潤程度均減輕。三組大鼠的癥狀學分析模型組評分高于對照組和干預組。AQP1主要表達于大鼠鼻黏膜被覆上皮基底膜、血管內皮細胞膜和腺體細胞內,VEGF主要表達于小動、靜脈血管內皮細胞和腺體細胞內。AQP1與VEGF在模型組中經免疫組織化學和real-time PCR法檢測均高表達,干預組表達低于模型組。差異均具有統(tǒng)計學意義,且兩者具有正相關性。結論AQP1、VEGF在實驗性變應性鼻炎大鼠鼻粘膜的強表達,表明AQP1、VEGF參與了變應性鼻炎的發(fā)病過程,與組織的水腫、腺體的過度分泌可能有關。糖皮質激素可以明顯下調AQP1、VEGF的表達水平,提示糖皮質激素可能是通過改變AQP1、VEGF的表達,減少腺體的分泌和改善鼻塞癥狀。且AQP1與VEGF之間具有正相關性,但具體機制尚不明確。
[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.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R765.21

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