本文選題：單側(cè)輸尿管梗阻 + 腺苷　； 參考：《中南大學(xué)》2014年碩士論文

【摘要】：目的：缺血和缺氧是導(dǎo)致慢性腎臟疾病的重要原因。腺苷作為缺血、缺氧性腎臟纖維化的重要信號(hào)分子,也可能是慢性腎臟疾病的一個(gè)重要致病因子。本研究檢測(cè)了單側(cè)輸尿管梗阻后腎臟腺苷濃度的變化及其相關(guān)致纖維化因子的表達(dá),探索了腺苷信號(hào)通路與腎纖維化的相關(guān)性,旨在進(jìn)一步闡述慢性腎臟發(fā)病機(jī)制。 方法：選擇清潔級(jí)健康雄性SD大鼠32只,隨機(jī)分為兩組：假手術(shù)組(對(duì)照組)和單側(cè)輸尿管梗阻組(UUO組),每組各16只,分別于手術(shù)后第1、2、3、4周分批處死前抽血,處死后取腎臟樣本,每個(gè)時(shí)間點(diǎn)在兩組中各取4只。分別用蘇木素伊紅(HE)、Masson染色觀察各時(shí)間點(diǎn)大鼠腎組織病理變化以及腎間質(zhì)膠原纖維沉積情況,應(yīng)用圖像分析系統(tǒng)進(jìn)行半定量分析。采用高效液相色譜法(HPLC)檢測(cè)腎組織腺苷水平,Elisa測(cè)定試劑盒檢測(cè)血肌酐水平,實(shí)時(shí)定量PCR測(cè)定腎間質(zhì)TGF-β1和Ⅰ型前膠原基因mRNA的表達(dá)量,免疫組織化學(xué)方法測(cè)定腎間質(zhì)TGF-β1的蛋白表達(dá),圖像分析系統(tǒng)對(duì)免疫組化結(jié)果進(jìn)行半定量分析。 結(jié)果：1、單側(cè)輸尿管梗阻后,實(shí)驗(yàn)組腺苷濃度相比對(duì)照組顯著升高(P0.05),并且在實(shí)驗(yàn)后第一周達(dá)到高峰(P0.01),HE和Masson染色結(jié)果顯示隨著腺苷濃度顯著增加,腎單位破壞逐漸增多,腎間質(zhì)膠原沉積逐漸增加。2、在單側(cè)輸尿管梗阻后的四個(gè)時(shí)間點(diǎn),未見血肌酐水平顯著(P0.05)增加。3、實(shí)時(shí)PCR數(shù)據(jù)和免疫組化分析顯示,在單側(cè)輸尿管梗阻后的四個(gè)時(shí)間點(diǎn)的TGF-β1和Ⅰ型前膠原的mRNA顯著增加(P0.05),免疫組化分析表明在UUO后第二周,其在腎小管上皮細(xì)胞表達(dá)最多,然后隨著時(shí)間推移,其表達(dá)逐漸下降,并且隨著UUO時(shí)間的延長(zhǎng),TGF-β1的分布從腎小管上皮細(xì)胞向腎間質(zhì)轉(zhuǎn)移,進(jìn)而蔓延至腎小球。 結(jié)論：1、腎間質(zhì)纖維化過(guò)程中可能存在腎組織持續(xù)低氧,并導(dǎo)致組織內(nèi)腺苷水平升高；2、腺苷含量增高,加速了腎小管損傷、腎間質(zhì)纖維化的發(fā)生、發(fā)展；3、腺苷信號(hào)通路可能通過(guò)調(diào)節(jié)TGF-β1、 α1(Ⅰ)procollagen mRNA表達(dá),干預(yù)RIF的發(fā)展。
[Abstract]:Objective: ischemia and hypoxia are important causes of chronic renal disease. Adenosine, as an important signal molecule of ischemia and hypoxic renal fibrosis, may also be an important pathogenic factor of chronic renal disease. In this study, the changes of adenosine concentration and the expression of fibrosis factors in the kidney after unilateral ureteral obstruction were detected, and the correlation between adenosine signaling pathway and renal fibrosis was explored. Methods: Thirty-two healthy male SD rats of clean grade were randomly divided into two groups: sham operation group (control group) and UUO group with unilateral ureteral obstruction (16 rats in each group). Blood samples were taken from the kidney at the 1st and 2nd week after the operation and 4 weeks after the operation. 4 rats in each group were collected at each time point. The pathological changes of renal tissue and the deposition of renal interstitial collagen fibers in rats were observed by Masson staining. The semi-quantitative analysis was carried out by image analysis system. High performance liquid chromatography (HPLC) was used to detect adenosine level in renal tissue and serum creatinine level was detected by Elisa kit. The expression of TGF- 尾 1 and procollagen 鈪，

本文編號(hào)：1997865