本文關(guān)鍵詞： miR-4284 動(dòng)脈硬化閉塞　出處：《西南醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:探討微小RNA-4284(MicroRNA-4284簡(jiǎn)稱miR-4284)在下肢動(dòng)脈硬化閉塞癥(Arteriosclerosis obliterans,ASO)患者血漿和病變血管中的表達(dá)量變化特點(diǎn);探討在缺血清缺氧狀態(tài)下miR-4284在動(dòng)脈平滑肌細(xì)胞增殖、遷移及凋亡中的作用。方法:臨床試驗(yàn):收集西南醫(yī)科大學(xué)第一附屬醫(yī)院血管甲狀腺外科2014-2015年間收治的55例下肢ASO患者和54例健康對(duì)照者血漿,24例下肢ASO截肢患者的病變血管及24例外傷截肢患者正常血管。選取其中3對(duì)年齡及性別相近的下肢ASO患者和正常健康人血漿用microarray篩選出下肢ASO患者血漿與正常健康人血漿中相比較表達(dá)差異在2倍以上的24種MicroRNAs(mi RNAs),用實(shí)時(shí)熒光定量PCR檢測(cè)臨床55例下肢ASO患者和54例健康對(duì)照人群血漿中miR-4284的表達(dá)量,24例下肢ASO截肢患者病變血管和24例正常血管中miR-4284的表達(dá)量;按Fontaine分期將55例下肢ASO患者分為Ⅰ、Ⅱ、Ⅲ、Ⅳ期,用RT-qPCR分別檢測(cè)各期患者血漿中mi R-4284的表達(dá)量。細(xì)胞實(shí)驗(yàn):正常培養(yǎng)主動(dòng)脈平滑肌、臍靜脈內(nèi)皮細(xì)胞后,再進(jìn)行缺血清缺氧處理用以模擬臨床ASO患者下肢缺血缺氧的狀態(tài)。用高糖培養(yǎng)基培養(yǎng)臍靜脈內(nèi)皮細(xì)胞再進(jìn)行缺血清缺氧處理,用以模擬臨床ASO合并Ⅱ型糖尿病患者。采用RT-qPCR檢測(cè)各組細(xì)胞中mi R-4284的相對(duì)表達(dá)量;分別上調(diào)、下調(diào)miR-4284后通過cck8檢測(cè)細(xì)胞增殖、transwell檢測(cè)細(xì)胞遷移,流式檢測(cè)細(xì)胞凋亡,蛋白印跡檢測(cè)凋亡相關(guān)蛋白XIAP、BCL2、BAX的表達(dá)。結(jié)果:臨床試驗(yàn)顯示,與健康對(duì)照組相比下肢ASO患者血漿中miR-4284的表達(dá)量在Fontaine第Ⅰ期中就開始顯著升高,并在第Ⅱ、Ⅲ、Ⅳ期血漿中一直持續(xù)在高水平狀態(tài);下肢ASO患者病變血管與正常血管相比,病變血管中miR-4284的表達(dá)量也明顯升高;細(xì)胞試驗(yàn):臍靜脈內(nèi)皮細(xì)胞在低糖及高糖缺血清缺氧的條件下miR-4284的表達(dá)量均升高;主動(dòng)脈平滑肌細(xì)胞在缺血清缺氧的條件下miR-4284表達(dá)量降低;主動(dòng)脈平滑肌細(xì)胞在缺血清缺氧的條件下,上調(diào)、下調(diào)miR-4284,對(duì)細(xì)胞的增殖、遷移沒有太大影響;在不進(jìn)行缺血清缺氧處理的主動(dòng)脈平滑肌細(xì)胞中,上調(diào)、下調(diào)miR-4284對(duì)細(xì)胞凋亡沒有太大影響;在缺血清缺氧的主動(dòng)脈平滑肌細(xì)胞中上調(diào)miR-4284的表達(dá)對(duì)細(xì)胞的凋亡沒有太大影響,但下調(diào)miR-4284的表達(dá)能抑制主動(dòng)脈平滑肌細(xì)胞凋亡;抑制凋亡相關(guān)蛋白XIAP、BCL2在下調(diào)mi R-4284后表達(dá)量明顯升高,促凋亡相關(guān)蛋白BAX在下調(diào)miR-4284后表達(dá)量明顯降低,差異有統(tǒng)計(jì)學(xué)意義。結(jié)論:miR-4284在下肢ASO患者血漿和血管組織中表達(dá)量明顯升高;缺血清缺氧誘導(dǎo)主動(dòng)脈平滑肌細(xì)胞中miR-4284表達(dá)明顯降低,缺血清缺氧誘導(dǎo)人臍靜脈內(nèi)皮細(xì)胞中miR-4284表達(dá)明顯升高;miR-4284對(duì)平滑肌細(xì)胞增殖、遷移作用不大;下調(diào)miR-4284可以抑制平滑肌細(xì)胞凋亡,調(diào)控miR-4284有望成為靶向治療下肢ASO研究的新方向。
[Abstract]:Objective: to investigate the changes of micro#en0# expression in plasma and diseased vessels of patients with Arteriosclerosis obliterans obliterans obliterans (Arteriosclerosis obliterans), and to investigate the proliferation of miR-4284 in arterial smooth muscle cells under ischemia and hypoxia. Role of migration and apoptosis. Methods: clinical trial: 55 patients with lower extremity ASO and 54 healthy controls who were treated in vascular thyroid surgery, first affiliated hospital of southwest medical university from 2014 to 2015, 24 patients with lower extremity ASO amputation. Three pairs of patients with age and sex of lower extremity ASO patients and healthy people were selected to use microarray to screen the plasma of lower extremity ASO patients compared with healthy people. Compared with 24 species of MicroRNAs(mi RNAsN, the expression of miR-4284 was detected by real-time fluorescence quantitative PCR in 55 patients with lower extremity ASO and 54 healthy controls. There were 24 patients with ASO amputation of lower extremity and 24 patients with normal blood. The expression of miR-4284 in tube; 55 patients with lower extremity ASO were divided into 鈪，

本文編號(hào)：1505455