本文選題：特發(fā)性肺纖維化　切入點(diǎn)：EMT　出處：《濱州醫(yī)學(xué)院》2012年碩士論文

【摘要】：目的觀察miRNAs (let-7d、miR-140、miR-192)在肺纖維化大鼠中的表達(dá)變化,研究其對(duì)TGF-β1/Smad3信號(hào)通路的調(diào)控作用及其對(duì)下游靶基因表達(dá)的影響,探討miRNA在肺纖維化發(fā)病中的作用機(jī)制。 方法SD大鼠60只隨機(jī)分為對(duì)照組及模型組,BLM氣管注入法建立肺纖維化動(dòng)物模型。分別于7d、14d、28d處死大鼠10只留取肺組織,HE染色和masson染色動(dòng)態(tài)觀察肺組織病理學(xué)變化,電鏡觀察肺組織超微結(jié)構(gòu)的變化,羥脯氨酸堿水解法測(cè)定膠原的合成；免疫組化法測(cè)定α-SMA、HMGA2、Thy-1、膠原蛋白I的表達(dá)；Real-time RT-PCR檢測(cè)miRNAs (let-7d、miR-140、miR-192)和TGF-β1/Smad3通路相關(guān)基因的表達(dá)量；Western Blotting檢測(cè)HMGA2、E-鈣粘蛋白的表達(dá)。 結(jié)果HE染色顯示BLM組肺組織按時(shí)間順序呈現(xiàn)由炎癥至纖維化的動(dòng)態(tài)改變；masson染色和羥脯氨酸含量檢測(cè)顯示肺組織膠原纖維的含量隨肺組織纖維化的程度逐漸增加；電鏡下可見Ⅱ型細(xì)胞數(shù)目較前減少,纖維母細(xì)胞增生,膠原纖維明顯增多；免疫組化結(jié)果示BLM組肺組織HMGA2、α-SMA、膠原蛋白I的表達(dá)明顯增加而thy-1蛋白的表達(dá)逐步下降；Real-time RT-PCR結(jié)果顯示,Let-7d、miR-140的表達(dá)隨纖維化程度的加重逐步下降,而miR-192的的表達(dá)量逐漸上升,三種miRNA的表達(dá)在肺纖維初期即變化明顯；TGF-β1、ZEB1、ZEB2, α-SMA的表達(dá)的升高,E-Cad基因mRNA的逐步下降。Western Blot結(jié)果示HMGA2的表達(dá)上升明顯、E-鈣粘蛋白逐步下降。 結(jié)論miRNA (let-7d、miR-140、miR-192)在肺纖維中通過smad3信號(hào)通路對(duì)EMT過程進(jìn)行調(diào)控。
[Abstract]:Objective to observe the expression of miRNAs let-7dnmiR-140 miR-1922 in rats with pulmonary fibrosis, to study its regulatory effect on TGF- 尾 1/Smad3 signaling pathway and its effect on downstream target gene expression, and to explore the mechanism of miRNA in the pathogenesis of pulmonary fibrosis.Methods 60 SD rats were randomly divided into control group and model group.Ten rats were killed on day 14 and 28, respectively. The pathological changes of lung tissue were observed by HE staining and masson staining. The ultrastructure of lung tissue was observed by electron microscope, and collagen synthesis was measured by hydrolysate of hydroxyproline.The expression of 偽 -SMA-HMGA _ 2 Thy-1, real-time RT-PCR of collagen I, miRNAs let-7dnmiR-140 miR-1922) and TGF- 尾 1/Smad3 pathway related genes were detected by immunohistochemistry. The expression of HMGA _ 2 and E-cadherin was detected by Western Blotting.Results HE staining showed that lung tissue in BLM group showed dynamic changes from inflammation to fibrosis in chronological order. Mass on staining and hydroxyproline content detection showed that the content of collagen fibers in lung tissue increased with the degree of pulmonary fibrosis.Under electron microscope, the number of type 鈪，

本文編號(hào)：1705719