本文選題：分泌性中耳炎 + 水通道蛋白��； 參考：《東南大學(xué)》2016年碩士論文

【摘要】：目的：探究模型豚鼠在分泌性中耳炎(Otitis media with effusion, OME)自然轉(zhuǎn)歸過(guò)程中,咽鼓管黏膜上皮內(nèi)AQP1、AQP5的表達(dá)變化情況,試圖從新的角度探討AQP參與調(diào)控中耳液體平衡的機(jī)制。方法：健康雄性豚鼠35只,采用左側(cè)咽鼓管阻塞法建立可逆性O(shè)ME豚鼠模型,耳內(nèi)鏡下觀察并記錄造模成功豚鼠的鼓膜形態(tài)及鼓室積液情況,將造模成功的豚鼠隨機(jī)分為“術(shù)后3d組”、“術(shù)后7d組”、“術(shù)后14d組”和“術(shù)后28d組”四個(gè)實(shí)驗(yàn)組,分別于術(shù)后第3天、第7天、第14天和第28天處死、采集標(biāo)本。同時(shí)取5只健康雄性豚鼠的左側(cè)咽鼓管組織作為對(duì)照。蘇木素-伊紅(HE)染色觀察OME豚鼠咽鼓管黏膜的組織學(xué)形態(tài)變化,免疫組織化學(xué)染色觀察AQP1、AQP5在OME豚鼠模型咽鼓管黏膜上皮內(nèi)的表達(dá)部位,蛋白免疫印跡法檢測(cè)各組中AQP1,5的表達(dá)量是否有差異。結(jié)果：27只實(shí)驗(yàn)豚鼠造模成功,術(shù)后每三天耳內(nèi)鏡下觀察并記錄鼓膜形態(tài)及鼓室積液情況,將中耳感染及死亡的豚鼠排除實(shí)驗(yàn)隊(duì)列,最終“術(shù)后3d組”、“術(shù)后7d組”、“術(shù)后14d組”和“術(shù)后28d組”符合實(shí)驗(yàn)要求予以納入的實(shí)驗(yàn)豚鼠例數(shù)為6例、5例、4例、3例。HE染色觀察到對(duì)照組的咽鼓管黏膜纖毛排列整齊,形態(tài)完整；而“術(shù)后3d組”和“術(shù)后7d組”的咽鼓管黏膜出現(xiàn)纖毛紊亂、壞死、缺失,上皮層及固有層水腫,混合性腺體病理性增生等形態(tài)學(xué)改變。隨著疾病的自動(dòng)轉(zhuǎn)歸,在“術(shù)后14d組”和“術(shù)后28d組”中可以觀察到咽鼓管黏膜的纖毛形態(tài)逐漸恢復(fù),黏膜水腫較前減輕。免疫組織化學(xué)染色觀察到AQP1表達(dá)于咽鼓管黏膜下的成纖維細(xì)胞以及血管內(nèi)皮細(xì)胞；AQP5表達(dá)于咽鼓管黏膜的纖毛細(xì)胞頂部以及混合性腺體。蛋白免疫印跡實(shí)驗(yàn)顯示AQP1與AQP5的表達(dá)量在“術(shù)后3d組”出現(xiàn)了顯著降低,“術(shù)后7d組”維持在較低水平,而在“術(shù)后14d組”出現(xiàn)了表達(dá)上升,“術(shù)后28d組”中的表達(dá)則進(jìn)一步增加。結(jié)論：1.AQP1和AQP5在豚鼠中耳咽鼓管黏膜內(nèi)存在表達(dá),其中AQP1表達(dá)于咽鼓管黏膜下的成纖維細(xì)胞以及血管內(nèi)皮細(xì)胞；AQP5表達(dá)于咽鼓管黏膜的纖毛細(xì)胞頂部以及混合性腺體。2.咽鼓管黏膜內(nèi)的AQP1和AQP5在OME發(fā)病以及自然轉(zhuǎn)歸的過(guò)程中出現(xiàn)了表達(dá)明顯下降隨后緩慢上升的變化,并且AQP1和AQP5的表達(dá)水平與咽鼓管黏膜的受損程度存在關(guān)聯(lián)。3.咽鼓管黏膜內(nèi)的AQP1和AQP5對(duì)于維持咽鼓管黏膜的完整性和黏液纖毛系統(tǒng)的正常運(yùn)作具有重要意義,并可能通過(guò)該途徑參與了中耳液體平衡的調(diào)節(jié)以及OME的發(fā)生、發(fā)展。
[Abstract]:Objective: To explore the changes in the expression of AQP1 and AQP5 in the epithelia of the eustachian tube during the natural outcome of Otitis media with effusion (OME) in the model guinea pig, and to explore the mechanism of AQP involved in regulating the liquid balance of the middle ear from a new angle. Methods: 35 healthy male guinea pigs were established by the left eustachian tube obstruction method. In a reversible OME guinea pig model, the tympanic membrane morphology and tympanic cavity effusion were observed and recorded by endoscopy. The guinea pigs were randomly divided into four groups, "postoperative 3D group", "postoperative 7d group", "postoperative 14d group" and "postoperative 28d group", respectively, to be killed at third days, seventh days, fourteenth days and twenty-eighth days after operation respectively. At the same time, the left eustachian tube tissue of 5 healthy male guinea pigs was taken as control. The histological changes of the eustachian tube mucosa of OME guinea pigs were observed by hematoxylin eosin (HE) staining. The expression of AQP1, AQP5 in the mucosa of the eustachian tube of OME guinea pig was observed by immunohistochemical staining, and AQP1,5 was detected by protein immunoblotting in the OME guinea pig model. Results: 27 experimental guinea pigs were made successfully. The morphology of the tympanic membrane and the effusion of tympanic cavity were recorded every three days after ear endoscopy. The middle ear infection and the dead guinea pigs were excluded from the experimental cohort, and the final "postoperative 3D group", "postoperative 7d group", "postoperative 14d group" and "postoperative 28d group" were accorded with the experimental requirements. The number of experimental guinea pigs included 6 cases, 5 cases, 4 cases. 3 cases of.HE staining showed that the mucociliary mucosa of the eustachian tube in the control group was neatly arranged, and the morphology of the eustachian tube in the post operation 3D group and the 7d group after operation appeared ciliary disorder, necrosis, deletion, epithelium and lamina edema, and the pathological hyperplasia of mixed gland. In the "postoperative 14d group" and "postoperative 28d group", the cilium morphology of the eustachian tube was gradually recovered and the edema of the mucous membrane decreased. Immunohistochemical staining was used to observe the fibroblasts and intravascular cells expressed under the mucosa of the eustachian tube by immunohistochemical staining. AQP5 was expressed in the cilium of the eustachian tube mucosa. The expression of AQP1 and AQP5 showed a significant decrease in the "postoperative 3D" group. "Post operation 7d group" was maintained at a lower level, while the expression in "postoperative 14d group" increased, and the expression in group 28d was further increased. Conclusion: 1.AQP1 and AQP5 are in guinea pigs. The expression of AQP1 was expressed in the mucous membrane of the eustachian tube, in which the fibroblasts and vascular endothelial cells were expressed under the mucous membrane of the eustachian tube; the AQP1 and AQP5 expressed in the ciliated cells of the eustachian tube and the AQP1 and AQP5 in the mixed.2. eustachian tube mucosa showed a significant decline in the expression of the OME and the natural outcome of the eustachian mucosa. The expression level of AQP1 and AQP5 and the damage degree of the eustachian tube mucosa are associated with the AQP1 and AQP5 in the.3. eustachian tube mucosa. It is important to maintain the integrity of the eustachian tube mucosa and the normal operation of the mucociliary system, and may be involved in the regulation of liquid balance in the middle ear and OME. Happen, develop.

【學(xué)位授予單位】：東南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R764.21;R-332

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本文編號(hào)：1779716