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  本文關鍵詞： 深靜脈血栓 Kr(u|¨)ppel樣因子4 分子標記物　出處：《昆明醫(yī)科大學》2012年碩士論文　論文類型：學位論文

【摘要】：目的： 在前期對形成DVT大鼠股靜脈動物模型Affymetrix Rat230A cDNA基因芯片的統(tǒng)計研究,篩查歸納深靜脈血栓形成時期具有明顯差異性表達的基因的基礎上,通過查閱PubMand和Gene Card數據庫,鎖定深靜脈血栓形成下調基因KLF-4。檢測KLF-4在正常人血液樣本、血栓未形成患者血液樣本及深靜脈血栓形成患者血液樣本中表達變化,探討KLF-4的表達情況與深靜脈血栓形成的關系,確定人血細胞轉錄層面KLF-4作為早期診斷人深靜脈血栓的候選分子標志物。 方法： 1、參考《靜脈血栓栓塞癥預防的NICE指南》(2012年)和《美國胸科醫(yī)師協(xié)會抗栓與血栓預防臨床實踐指南—深靜脈血栓形成的診斷》(2012年第9版)制定本研究的DVT診斷標準：DVT的綜合診斷=危險因素評估(基礎危險因素+專科危險因素)+主要臨床表現+下肢深靜脈彩色多普勒超聲檢查(附錄2)。于2010年3月-2012年3月共納入樣本數58例,正常對照組(20例,為健康志愿者)、血栓未形成組(20例,為下肢長骨骨折或脊柱骨折7d內行手術后無形成血栓的患者)、血栓形成組(18例,為DVT形成患者),三組均采集清晨空腹靜脈血液樣本,血栓未形成組于手術后第1d采集,血栓形成組于確診后第1d采集。收集血樣本,應用primer5.0引物設計軟件對人KLF-4設計反轉錄引物,將各組人血細胞總RNA反轉錄為cDNA,以GAPDH為內參照運用半定量PCR檢測KLF-4在臨床血液樣本白細胞中的表達情況。 2、對實驗數據進行處理,分析KLF-4在深靜脈血栓形成中的表達情況和作用,確定KLF-4在人血細胞轉錄層面能夠作為早期診斷DVT及預后轉歸的候選分子標志物。 結果 從半定量PCR膠圖及目的基因與內參基因灰度值比值可以看出：內參對照GAPDH大小均一,亮度一致,C組條帶明顯較A組及B組條帶亮度降低,灰度值：A組(0.9803±0.0740),B組(0.9764±0.0839)C組(0.3751±0.0359),C組與A組相比P0.05,差距有統(tǒng)計學意義,C組與B組比較P0.05,差距有統(tǒng)計學意義,B組與A組相比P0.05,差距無統(tǒng)計學意義。 結論 1.KLF-4與深靜脈血栓之間存在著非常密切的相關性,很有可能為影響深靜脈血栓形成與否的一個主要影響因素。 2.KLF-4在人深靜脈血栓患者血液中的表達下調,可以作為分子標記物在血液中進行檢測來診斷人深靜脈血栓。
[Abstract]:Objective:. On the basis of the statistical study of Affymetrix Rat230A cDNA gene chip in the animal model of femoral vein formation in DVT rats, and the screening of genes with obvious difference expression during deep venous thrombosis, the databases of PubMand and Gene Card were consulted. To investigate the relationship between the expression of KLF-4 and deep venous thrombosis (DVT), we detected the expression of KLF-4 in the blood samples of normal subjects, non-thrombosis patients and deep venous thrombosis patients, and investigated the relationship between the expression of KLF-4 and deep venous thrombosis (DVT). To identify human blood transcriptional KLF-4 as a candidate molecular marker for early diagnosis of human deep venous thrombosis. Methods:. 1. Refer to the NICE guidelines for the Prevention of Venous Thromboembolism (2012) and the American Association of chest Physicians' guidelines for Clinical practice on Antithrombotic and Thrombosis Prevention-diagnosis of Deep Venous Thrombosis (2012 9th Edition) to develop the DVT diagnostic criteria for this study. The main clinical manifestations of quasi: DVT: comprehensive diagnosis = risk factor assessment (basic risk factors specialist risk factors) (appendix 2). From March 2010 to March 2012, 58 samples were included. The normal control group consisted of 20 healthy volunteers, 20 patients without thrombosis, 18 patients with thrombus after 7 days of operation and 18 patients with long bone fracture or spinal fracture of lower extremity. For the patients with DVT, fasting venous blood samples were collected in all three groups. Blood samples were collected on the first day after operation in the thrombus group, and on the first day after the diagnosis in the thrombosis group. Primer5.0 primer design software was used to design reverse transcription primers for human KLF-4. The total RNA of human blood cells in each group was reversed to cDNA. using GAPDH as internal reference, semi-quantitative PCR was used to detect the expression of KLF-4 in the leukocytes of clinical blood samples. 2. The experimental data were processed, and the expression and role of KLF-4 in deep venous thrombosis were analyzed. It was confirmed that KLF-4 could be used as a candidate molecular marker for early diagnosis and prognosis of DVT at the transcriptional level of human blood cells. Results. From the semi-quantitative PCR gel map and the ratio of gray value of target gene to internal reference gene, it can be seen that the size of GAPDH in the control group is uniform, and the brightness of the band in group C is obviously lower than that in group A and group B. The grayscale value of group A was 0.9803 鹵0.0740, group B was 0.9764 鹵0.0839C, the difference between group C and group A was 0.3751 鹵0.0359, the difference between group C and group A was significant (P 0.05), the difference between group B and group A was significant (P 0.05), the difference between group B and group A was not statistically significant. Conclusion. 1. KLF-4 is closely related to deep venous thrombosis, which may be one of the main factors influencing the formation of deep venous thrombosis. 2. The expression of KLF-4 was down-regulated in the blood of patients with deep venous thrombosis (DVT). KLF-4 could be used as a molecular marker to detect human deep venous thrombosis (DVT).
【學位授予單位】：昆明醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R363

【引證文獻】

相關博士學位論文 前1條

1 趙學凌;創(chuàng)傷性肢體深靜脈血栓形成的新型動物模型建立及相關研究[D];昆明醫(yī)學院;2004年

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本文編號：1522223