發(fā)布時間：2018-05-02 08:13

  本文選題：上皮間質(zhì)轉化 + 足細胞　； 參考：《山東大學》2017年碩士論文

【摘要】：在腎臟,足細胞附著在腎小球基底膜外,其足突上多種分子相互交錯形成裂孔隔膜(slit diaphragm,SD),是腎小球濾過屏障的重要組成部分。足細胞在保護腎小球濾過屏障的完整性方面起著重要的作用。足細胞的功能與腎臟功能密切相關,其超微結構的變化或與功能相關的分子表達改變均與腎臟功能的損害有著密切的關系。一般來說,上皮-間質(zhì)轉化(epithelial-mesenchymal transition,EMT)是極化上皮細胞發(fā)生許多生化變化,以獲得間充質(zhì)細胞表型。足細胞足上皮細胞,當受到不同的刺激(如TGF-β,高濃度的葡萄糖,血管緊張索Ⅱ,阿霉素等)時,有可發(fā)生EMT。當EMT的發(fā)生,上皮表型標志物如E-cadherin,P-cadherin,nephrin、podocalyxin,Synaptopodin 的下調(diào),與間質(zhì)表型標志物如desmin,FSP-1,MMP-9,ILK,fibronectin,vimentin,α-SMA,Ⅰ 型膠原及 snai 等上調(diào)。在受損的腎臟產(chǎn)生腎間質(zhì)纖維化中,EMT被認為是一個關鍵因素。足細胞EMT可引起腎小球濾過屏障的損傷和蛋白尿,嚴重者可導致足細胞由腎小球基底膜脫離和細胞凋亡,導致足細胞數(shù)量減少,從而加重蛋白尿和腎臟功能損傷甚至腎小球硬化。足細胞EMT是研究蛋白尿和腎臟纖維化發(fā)生發(fā)展的一個重要方面。研究目的:探討在高濃度的葡萄糖和血管緊張素Ⅱ存在下,足細胞的EMT以及TCF8在足細胞EMT中的影響。方法:足細胞經(jīng)分化后,經(jīng)與血管緊張素Ⅱ和葡萄糖分別孵育,Western blotting方法檢測蛋白表達;Transwell小室檢測細胞的遷移和侵襲能力。qPCR方法檢測mRNA表達。結果:1.血管緊張素Ⅱ和高糖對正常足細胞E-cadherin、α-catenin、N-cadherin和vimentin蛋白表達的影響血管緊張素Ⅱ和葡萄糖可使足細胞上皮標志分子E-cadherin和α-catenin蛋白的表達逐漸降低,而足細胞間質(zhì)標志分子N-cadherin和Vimentin表達均升高。2.血管緊張素Ⅱ和高糖對正常足細胞TCF8蛋白表達的影響足細胞經(jīng)葡萄糖或血管緊張素Ⅱ孵育后,隨著葡萄糖或血管緊張素Ⅱ濃度的增加,TCF8蛋白的表達逐漸升高。3.血管緊張素Ⅱ和高糖對正常足細胞遷移和侵襲能力足細胞經(jīng)葡萄糖或血管緊張素Ⅱ作用后,其遷移和侵襲能力有明顯增加。4.TCF8 siRNA 篩選足細胞經(jīng)TCF8基因沉默后,TCF8的表達明顯降低,其中1#siRNA的作用最強。5.沉默 TCF8 基因?qū)?E-cadherin、a-catenin、N-cadherin 和 vimentin 蛋白表達的影響未經(jīng)轉染的細胞上皮標志物分子E-cadherin和α-catenin均有表達,而間質(zhì)標志物分子N-cadherin和vimentin較低。經(jīng)高糖或血管緊張素Ⅱ刺激后,E-cadherin 和 α-catenin 表達降低,N-cadherin 和 vimentin 表達增加。shTCF8#1轉染細胞后,高濃度葡萄糖和血管緊張素Ⅱ均可使上皮細胞標志物E-cadherin和α-catenin表達增加,間質(zhì)標志物分子N-cadherin和vimentin表達降低,表明TCF8表達沉默后,細胞不利于間質(zhì)轉換,提示TCF8 具有促進EMT作用。6.沉默TCF8基因?qū)ψ慵毎w移和侵襲能力未經(jīng)轉染的細胞經(jīng)高糖或血管緊張素Ⅱ刺激后,細胞遷移和侵襲能力增加,TCF8基因沉默后,細胞遷移和侵襲能力降低。表明TCF8表達沉默后,細胞遷移能力下降。結論:本研究結果提示高糖和血管緊張素Ⅱ可促進足細胞EMT,TCF8促進了該EMT過程。
[Abstract]:In the kidneys, the podocytes are attached to the glomerular basement membrane, and a variety of molecules on the Poddar interlock to form slit diaphragm (SD). It is an important part of the glomerular filtration barrier. Podocyte plays an important role in protecting the integrity of the glomerular filtration barrier. The function of the podroblast is closely related to the function of the kidney. Changes in ultrastructure or changes in molecular expression related to function are closely related to impairment of renal function. Generally, epithelial-mesenchymal transition (EMT) is a variety of biochemical changes in the polarized epithelial cells in order to obtain the phenotype of mesenchymal cells. Stimulation (such as TGF- beta, high concentration of glucose, angiotensin II, adriamycin, etc.) can occur when EMT. occurs when EMT occurs, the epithelial phenotype markers such as E-cadherin, P-cadherin, nephrin, podocalyxin, Synaptopodin, and interstitial markers such as desmin, FSP-1, MMP-9, ILK, alpha, type I collagen and EMT is considered to be a key factor in the production of renal interstitial fibrosis in damaged kidneys. Podocyte EMT can cause damage to the glomerular filtration barrier and proteinuria. The serious person can cause the foot cell to be separated from the glomerular basement membrane and apoptosis, which leads to the decrease of the number of podons, thus aggravating the damage of proteinuria and renal function. To glomerulosclerosis. Podocyte EMT is an important aspect of the development of proteinuria and renal fibrosis. The purpose of this study is to explore the effect of EMT and TCF8 on the EMT of podocytes in the presence of high concentration of glucose and angiotensin II, and the effect of the podocyte on the foot cell EMT. The protein expression was detected by Western blotting method, and the cell migration and invasion ability of Transwell cells was detected by.QPCR method to detect mRNA expression. Results: 1. angiotensin II and high glucose affect the expression of E-cadherin, alpha -catenin, N-cadherin and vimentin protein in normal poddin, and the angiotensin II and glucose can make the foot cell epithelia standard The expression of E-cadherin and alpha -catenin protein decreased gradually, while the expression of N-cadherin and Vimentin in the stromal marker molecules increased the expression of.2. angiotensin II and high glucose on the expression of TCF8 protein in normal podocytes. After incubation of glucose or angiotensin II, the concentration of glucose or angiotensin II increased. Adding, the expression of TCF8 protein increased gradually,.3. angiotensin II and high glucose had the ability to migrate and invasiveness of normal poddcells. After the action of glucose or angiotensin II, the ability to migrate and invasiveness increased significantly in.4.TCF8 siRNA screening podthe cells after TCF8 gene silencing, and the expression of TCF8 was significantly reduced, of which 1#siRNA was most effective. The expression of E-cadherin, a-catenin, N-cadherin and vimentin protein expressed by the strong.5. silencing gene were expressed in the untransfected cell epithelial marker molecules E-cadherin and alpha -catenin, while the molecules N-cadherin and vimentin were lower in the interstitial markers. The expression of E-cadherin and alpha -catenin was reduced after the stimulation of high glucose or angiotensin II. Low, N-cadherin and vimentin expression increased.ShTCF8#1 transfected cells. High concentration of glucose and angiotensin II could increase the expression of E-cadherin and alpha -catenin in epithelial cell markers. The expression of N-cadherin and vimentin in the interstitial marker molecules decreased, indicating that after the expression of TCF8 is silent, the cells are not conducive to interstitial conversion, suggesting that TCF8 is promoted. The effect of.6. silencing of TCF8 gene on the cell migration and invasion of untransfected cells with high glucose or angiotensin II, increased cell migration and invasion ability, and decreased cell migration and invasion ability after TCF8 gene silencing. It indicated that the cell migration ability decreased after the silent TCF8 gene was silent. Conclusion: the results of this study suggest that high glucose is the result of this study. And angiotensin II can promote podocyte EMT and TCF8 promote the EMT process.

【學位授予單位】：山東大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R692

【相似文獻】

相關期刊論文 前10條

1 張玉珍,高平進,王興宇,張宗梁,朱鼎良;血管緊張素Ⅱ?qū)ρ芷交〖毎鲋车挠绊慬J];中國病理生理雜志;2000年10期

2 張怡;血管緊張素Ⅱ-2型受體的最新研究進展[J];心血管病學進展;2001年05期

3 吳自強;血管緊張素Ⅱ的生成機制[J];臨床內(nèi)科雜志;2003年07期

4 沈云輝,陳長勛;血管緊張素Ⅱ的生物學效應及中藥對其影響[J];中國中藥雜志;2003年05期

5 陳世德;血管緊張素Ⅱ與纖溶酶原激活物抑制物-1的研究進展[J];醫(yī)學綜述;2003年09期

6 白敬恩,李榮山;血管緊張素Ⅱ與紅系造血[J];國外醫(yī)學.泌尿系統(tǒng)分冊;2005年02期

7 鄧曉奇;;低密度脂蛋白膽固醇對血管緊張素Ⅱ敏感性的影響[J];心血管病學進展;2006年04期

8 關愛麗;牛玉宏;張國平;李紀明;馬楨;梁艷艷;胡朝輝;鄒云增;;血管緊張素Ⅱ引起心肌微血管內(nèi)皮細胞功能障礙及其可能機制的研究[J];中國分子心臟病學雜志;2009年01期

9 劉迎春;李慧敏;劉U，

本文編號：1833099