本文選題：DDR2　切入點(diǎn)：肺纖維化　出處：《第四軍醫(yī)大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：肺纖維化是由多種因素引起的慢性肺間質(zhì)性疾病。由于工業(yè)的高速發(fā)展、吸煙人群以及老齡化人口的增加，肺纖維化已成為我國的常見病和多發(fā)病。盡管科學(xué)家對此疾病已經(jīng)進(jìn)行了大量的研究，但到目前為止，肺纖維化的病因和發(fā)病機(jī)制仍然不甚清楚。早期診斷困難、預(yù)后差以及缺乏有效的治療手段使得肺纖維化成為難以攻克的疾病，亟待更深入的研究。 DDR2分子屬于酪氨酸蛋白激酶受體（Receptor Tyrosine Kinase，RTK）家族成員，介導(dǎo)細(xì)胞與細(xì)胞外基質(zhì)之間的信號轉(zhuǎn)導(dǎo)，廣泛參與細(xì)胞增殖分化、個體發(fā)育及生殖等生理病理過程。當(dāng)配體與DDR2結(jié)合后，其胞內(nèi)區(qū)的酪氨酸殘基發(fā)生磷酸化。磷酸化的位點(diǎn)會進(jìn)一步招募下游的信號分子，最終導(dǎo)致整個信號通路的活化。其活化的持續(xù)時間較經(jīng)典RTK明顯提高，提示DDR2可能參與體內(nèi)較緩慢的病理過程。 已有研究表明，DDR2僅在肺、胃等臟器中高表達(dá)。而肺纖維化過程中產(chǎn)生的大量膠原，特別是I型和III型膠原，，為活化DDR2提供了空間上的可能；在人原發(fā)性膽汁性肝硬變和大鼠酒精性肝纖維化模型中，DDR2的表達(dá)水平隨著病程的發(fā)展而逐漸升高；DDR2還能通過調(diào)控膠原酶MMP-1和MMP13的表達(dá)參與類風(fēng)濕性關(guān)節(jié)炎的發(fā)生發(fā)展過程，而MMPs正好是肺纖維化的重要參與分子。因此，我們推測DDR2在肺纖維化過程中必然發(fā)揮重要作用。 我們主要進(jìn)行了以下幾方面的研究： 1.在人正常肺標(biāo)本和肺纖維化標(biāo)本中通過免疫熒光方法檢測DDR2的表達(dá)差異。 2.建立小鼠Bleomycin肺纖維化模型，實(shí)時定量PCR和Western Blot方法檢測肺纖維化過程中的DDR2表達(dá)變化。 3.在DDR2基因缺失小鼠上建立肺纖維化動物模型，HE染色、Masson染色、ELISA、免疫熒光、TUNEL、明膠酶譜等方法明確DDR2在肺纖維過程中的作用。 4. siDDR2特異性下調(diào)野生鼠肺組織DDR2的表達(dá)，采用HE、Masson等方法觀察DDR2被干涉之后肺纖維化水平變化。 5. TGF-β1刺激小鼠原代肺成纖維細(xì)胞，實(shí)時定量PCR和Western Blot方法檢DDR2信號阻斷后纖維化相關(guān)分子以及下游信號分子的表達(dá)變化。 6.在肺纖維化模型上，觀察DDR2特異性小分子抑制劑對肺纖維化的影響。 結(jié)果：人肺纖維化標(biāo)本較正常肺標(biāo)本DDR2表達(dá)水平明顯升高；小鼠肺纖維化過程中DDR2表達(dá)呈現(xiàn)先下降后上升的趨勢；DDR2基因缺失后炎癥侵潤現(xiàn)象明顯減輕，MMPs表達(dá)和分泌下降；DDR2野生鼠中肺纖維化水平明顯升高，肺組織羥脯氨酸含量和肺泡灌洗液中TGF-β1的量明顯高于DDR2基因缺失鼠，纖維化晚期凋亡細(xì)胞也較野生鼠明顯減少；細(xì)胞學(xué)實(shí)驗表明肺纖維化相關(guān)分子的表達(dá)在DDR2缺失后隨著TGF-β1誘導(dǎo)時間的延長顯著下降，AKT、p38等下游信號分子磷酸化水平和肌成纖維細(xì)胞數(shù)目也顯著下降。
[Abstract]:Pulmonary fibrosis is a chronic pulmonary interstitial disease caused by many factors. Pulmonary fibrosis has become a common disease and frequent disease in China. Although scientists have done a lot of research on this disease, the etiology and pathogenesis of pulmonary fibrosis are still unclear. Poor prognosis and lack of effective treatment make pulmonary fibrosis a difficult disease. DDR2 molecule is a member of the tyrosine protein kinase receptor receptor receptor (DDR2). It mediates the signal transduction between cells and extracellular matrix, and participates in many physiological and pathological processes, such as cell proliferation, differentiation, ontogenesis and reproduction. When ligand binds to DDR2, it plays an important role in the process of cell proliferation, differentiation, ontogenesis and reproduction. The phosphorylation of tyrosine residues in the intracellular region may further recruit downstream signaling molecules and eventually lead to activation of the entire signaling pathway. The duration of activation is significantly longer than that of classical RTK. The results suggest that DDR2 may be involved in the slower pathological process in vivo. It has been shown that DDR2 is only highly expressed in the lung and stomach. However, the large amount of collagen produced in the process of pulmonary fibrosis, especially type I and III collagen, provides a spatial possibility for activating DDR2. The expression of dddR2 in human primary biliary cirrhosis and alcoholic liver fibrosis model increased gradually with the development of the course of the disease. The expression of DDR2 also participated in the pathogenesis of rheumatoid arthritis by regulating the expression of collagenase MMP-1 and MMP13. Therefore, we speculate that DDR2 must play an important role in the process of pulmonary fibrosis. We have mainly carried out the following studies:. 1. The expression of DDR2 was detected by immunofluorescence in normal lung samples and pulmonary fibrosis specimens. 2. The mouse Bleomycin pulmonary fibrosis model was established, and the expression of DDR2 in pulmonary fibrosis was detected by real-time quantitative PCR and Western Blot. 3. The animal model of pulmonary fibrosis was established in mice with DDR2 gene deletion. The role of DDR2 in the process of pulmonary fiber was clarified by means of HE staining and Masson staining, immunofluorescence Tunel and gelatinase spectrum. 4. SiDDR2 specifically down-regulated the expression of DDR2 in the lung tissues of wild rats, and observed the changes of pulmonary fibrosis after DDR2 was interfered with. 5. TGF- 尾 1 stimulated primary mouse lung fibroblasts, and real-time quantitative PCR and Western Blot were used to detect the expression of fibrosis related molecules and downstream signal molecules after DDR2 signal was blocked. 6. To observe the effect of DDR2 specific small molecule inhibitor on pulmonary fibrosis model. Results: the expression of DDR2 in human lung fibrosis was significantly higher than that in normal lung. In the course of pulmonary fibrosis in mice, the expression of DDR2 decreased first and then increased. The expression and secretion of DDR2 in the wild mice decreased significantly after the absence of DDR2 gene. The contents of hydroxyproline in lung tissue and TGF- 尾 1 in alveolar lavage fluid were significantly higher than those in mice without DDR2 gene, and the apoptotic cells in the late stage of fibrosis were significantly lower than those in wild mice. Cytological experiments showed that the expression of pulmonary fibrosis related molecules decreased significantly with the prolongation of TGF- 尾 1 induction time after DDR2 deletion, and the phosphorylation level of downstream signal molecules such as AKTTN p38 and the number of myofibroblasts also decreased significantly.
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R563.9

【共引文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 張燕;DDR2在成骨細(xì)胞分化過程中的作用研究[D];第四軍醫(yī)大學(xué);2010年

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