本文選題：水通道蛋白 + 小腸　； 參考：《吉林農(nóng)業(yè)大學》2017年碩士論文

【摘要】：水通道蛋白(Aquaporin,AQP)是廣泛存在于原核和真核細胞膜上快速轉(zhuǎn)運水分子的特異性孔道蛋白,目前已經(jīng)從哺乳動物中克隆出13種水通道蛋白(AQP0-AQP12)。研究表明水通道蛋白在機體體液轉(zhuǎn)運生理及病理過程(包括尿濃縮、外分泌腺功能、眼球房水代謝、腦水腫形成以及腦脊液分泌和吸收)中發(fā)揮重要作用。腸道作為動物體內(nèi)僅次于腎臟的第二大水鹽代謝器官,水通道的表達和功能的研究一直是該領域國際研究的前沿和熱點。然而,截至目前對于腸道中水通道的研究主要集中在人和大鼠結(jié)腸,而關于小鼠腸道,特別是小腸各段水通道蛋白的表達和功能的研究卻鮮有報道。本研究首先應用PCR和免疫印跡技術檢測正常小鼠十二指腸、空腸和回腸中水通道蛋白的表達情況。結(jié)果顯示AQP3在小鼠空腸中存在基因和蛋白表達,AQP4在小鼠回腸中存在基因和蛋白表達。為進一步明確這兩種水通道蛋白在小腸中的表達位置,我們通過免疫組織化學的方法證實,AQP3主要表達在小鼠空腸黏膜上皮細胞,而AQP4主要表達在小鼠回腸隱窩細胞基底膜。產(chǎn)腸毒素型大腸桿菌(Enterotoxigenic Escherichia coli,ETEC)是引起人和動物,特別是低齡動物腹瀉的主要病原菌之一。目前對于ETEC的致病機理的研究主要集中在其產(chǎn)生的兩種腸毒素(LT和ST型)通過影響小腸上皮細胞CFTR的功能,加強小腸對離子的分泌而導致腹瀉。但是,ETEC對于小腸上皮細胞水轉(zhuǎn)運功能的影響,目前還是研究的盲點。本研究通過建立ETEC誘導的小鼠腹瀉模型,應用RT-PCR、qPCR、ELISA等實驗方法檢測了感染ETEC產(chǎn)生腹瀉后小鼠AQPs基因和蛋白表達量的變化。結(jié)果顯示,小鼠感染ETEC導致腹瀉后,空腸中AQP3和回腸中AQP4的基因及蛋白表達量與正常對照組相比都明顯下降。因此我們推測,ETEC附著小腸后,可能通過釋放腸毒素使AQP3和AQP4表達量降低,影響小腸上皮細胞的水轉(zhuǎn)運功能,從而引起腹瀉。
[Abstract]:Aquaporin aqup is a special pore protein which widely transports water molecules on prokaryotic and eukaryotic cell membranes. At present, 13 aquaporin (AQP0-AQP12) proteins have been cloned from mammals. Studies have shown that aquaporins play an important role in the physiological and pathological processes of body fluid transport, including urine concentration, exocrine gland function, aqueous humor metabolism, cerebral edema formation, and cerebrospinal fluid secretion and absorption. Intestinal tract is the second largest water-salt metabolic organ in animals after kidney. The research on the expression and function of water channel has been the frontier and hotspot in this field. However, up to now, the study of the water channel in the intestinal tract is mainly focused on human and rat colon, but there are few reports on the expression and function of aquaporin in the intestine of mice, especially in the small intestine. In this study, the expression of aquaporin in duodenum, jejunum and ileum of normal mice was detected by PCR and Western blot. The results showed that there was gene and protein expression of AQP3 in the jejunum of mice. AQP4 expressed genes and proteins in mouse ileum. In order to further clarify the expression of these two aquaporins in the small intestine, we confirmed by immunohistochemical method that AQP3 was mainly expressed in mouse jejunal mucosal epithelial cells, and AQP4 was mainly expressed in the basement membrane of mouse ileum crypt cells. Enterotoxigenic Escherichia coli (ETEC) is one of the main pathogens causing diarrhea in humans and animals, especially in young animals. At present, the pathogenesis of ETEC is mainly focused on the two enterotoxins (LT and St type), which affect the function of CFTR in intestinal epithelial cells and enhance the secretion of ions in the small intestine and lead to diarrhea. However, the effect of ETEC on the water transport function of intestinal epithelial cells is still a blind spot. In this study, the mouse diarrhea model induced by ETEC was established, and the changes of AQPs gene and protein expression in mice with diarrhea induced by ETEC were detected by RT-PCR- qPCR- Elisa. The results showed that the gene and protein expression of AQP3 in jejunum and AQP4 in ileum were significantly decreased in mice infected with ETEC. Therefore, we speculate that the expression of AQP3 and AQP4 may be decreased by releasing enterotoxin, which may affect the water transport function of intestinal epithelial cells and cause diarrhea.
【學位授予單位】：吉林農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S856.4

【參考文獻】

相關期刊論文 前2條

1 Alexandra Zahn;Christoph Moehle;Thomas Langmann;Robert Ehehalt;Frank Autschbach;Wolfgang Stremmel;Gerd Schmitz;;Aquaporin-8 expression is reduced in ileum and induced in colon of patients with ulcerative colitis[J];World Journal of Gastroenterology;2007年11期

2 吳力克,梁冰,孫英姿,逄金聚,熊德鑫,梁明;復合益生菌制劑“海生元”治療實驗性腸菌群失調(diào)癥療效機制探討[J];中國微生態(tài)學雜志;2003年04期

，

本文編號：2004309