【摘要】：目的應(yīng)用微小RNA(miRNA)基因表達(dá)譜芯片技術(shù)篩查miRNA在原發(fā)性高血壓(EH)合并心力衰竭患者血漿中的差異表達(dá),旨在進(jìn)一步探索EH合并心力衰竭發(fā)生發(fā)展的分子機(jī)制。方法收集50例來(lái)自醫(yī)院心血管內(nèi)科住院部的EH合并心力衰竭急性發(fā)作期患者作為實(shí)驗(yàn)組。隨機(jī)選取50例EH不合并心力衰竭患者做為對(duì)照組。測(cè)定受試者血脂、血壓、體質(zhì)量指數(shù)(BMI)、氨基末端腦鈉尿肽前體(NT-proBNP)水平,評(píng)估臨床癥狀和超聲心動(dòng)圖心功能情況,常規(guī)抽提總RNA,應(yīng)用CapitalBio公司miRNA芯片檢測(cè)miRNA表達(dá)譜,并對(duì)差異表達(dá)的miRNA進(jìn)行實(shí)時(shí)熒光定量聚合酶鏈反應(yīng)(qRT-PCR)驗(yàn)證,觀察相應(yīng)差異表達(dá)的miRNA與NT-proBNP的相關(guān)性。結(jié)果與對(duì)照組比較,實(shí)驗(yàn)組血脂、血壓、BMI差異無(wú)統(tǒng)計(jì)學(xué)意義(均P0.05)。實(shí)驗(yàn)組患者血漿miRNA表達(dá)譜中明顯差異表達(dá)的miRNA共有19個(gè),其中有12個(gè)miRNA上調(diào)(miR-142-3p、miR-196、miR-28、miR-208等),7個(gè)miRNA下調(diào)(miR-542、miR-199a、miR-21、miR-195等),與qRT-PCR結(jié)果基本相符。miR-142-3p、miR-196與NT-proBNP呈明顯正相關(guān)。與對(duì)照組比較,心功能Ⅱ級(jí)患者血漿miR-142-3p和miR-196表達(dá)水平分別升高(4.44±0.21)倍和(3.78±0.20)倍(均P0.05),在心功能Ⅲ級(jí)患者分別升高(10.09±0.19)倍和(9.88±0.25)倍(均P0.05),在心功能Ⅳ級(jí)患者分別升高(15.34±0.18)倍和(13.83±0.22)倍(均P0.05);miR-542、miR-199a與NT-proBNP呈明顯負(fù)相關(guān)。與對(duì)照組比較,心功能Ⅱ級(jí)患者血漿miR-542和miR-199a表達(dá)水平分別降低(91±11)%和(85±10)%(均P0.05),在心功能Ⅲ級(jí)患者分別降低(55±9)%和(51±15)%(均P0.05),在心功能Ⅳ級(jí)患者分別降低(24±8)%和(22±7)%(均P0.05)。結(jié)論 EH合并心力衰竭患者血漿中miRNA的表達(dá)有明顯改變,并隨著心力衰竭嚴(yán)重程度的增加而相應(yīng)改變,與NT-proBNP明顯相關(guān),提示部分血漿miRNA可作為預(yù)測(cè)EH合并心力衰竭患者疾病進(jìn)展的新型生物學(xué)標(biāo)志物。
[Abstract]:Objective to screen the differential expression of miRNA in plasma of patients with essential hypertension (EH) complicated with heart failure by microarray RNA (miRNA) gene expression microarray, in order to further explore the molecular mechanism of occurrence and development of EH with heart failure. Methods 50 patients with EH complicated with acute attack of heart failure were selected as experimental group. Fifty patients with EH without heart failure were randomly selected as control group. Serum lipids, blood pressure, body mass index (BMI),) levels of brain natriuretic peptide precursors (NT-proBNP) were measured. Clinical symptoms and echocardiographic cardiac function were evaluated. Total RNA, was extracted by routine extraction. CapitalBio miRNA chip was used to detect the miRNA expression profile, and real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to verify the differentially expressed miRNA. The correlation between the differentially expressed miRNA and NT-proBNP was observed. Results there was no significant difference in blood lipid, blood pressure and BMI between the experimental group and the control group (P 0.05). In the experimental group, 19 miRNA were significantly differentially expressed in the plasma miRNA expression profile, of which 12 miRNA were up-regulated (miR-142-3p,miR-196,miR-28,miR-208 et al) and 7 miRNA were down-regulated (miR-542,miR-199a,miR-21,). MiR-195 et al., which were consistent with the results of qRT-PCR. MiR-142-3p,miR-196 was positively correlated with NT-proBNP. Compared with the control group, the expression of miR-142-3p and miR-196 in patients with cardiac function grade 鈪，

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