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動(dòng)脈粥樣硬化患者血清miR-520b表達(dá)水平及相關(guān)機(jī)制

發(fā)布時(shí)間:2019-07-10 15:52
【摘要】:目的探討動(dòng)脈粥樣硬化患者血清miR-520b表達(dá)水平及相關(guān)作用機(jī)制。方法選取該院收治的動(dòng)脈粥樣硬化患者42例,收集同期42名健康老年人為對(duì)照。qRT-PCR檢測(cè)外周血清miR-520b表達(dá)水平。經(jīng)targetscan和miRanda數(shù)據(jù)庫預(yù)測(cè)importin 8(IPO8)為miR-520b的可能靶基因,通過雙熒光素酶報(bào)告基因檢測(cè)證實(shí);體外細(xì)胞實(shí)驗(yàn)采用Western印跡檢測(cè)miR-520b對(duì)人血管內(nèi)皮細(xì)胞IPO8表達(dá)的影響;CCK-8法和Tranwells實(shí)驗(yàn)檢測(cè)miR-520b對(duì)人血管平滑肌細(xì)胞增殖、遷移能力的影響。結(jié)果動(dòng)脈粥樣硬化患者血清miR-520b相對(duì)表達(dá)量低于健康老年人(P0.05)。雙熒光素酶報(bào)告基因檢測(cè)證實(shí)IPO8是miR-520b的直接作用靶基因;轉(zhuǎn)染miR-520b類似物人血管內(nèi)皮細(xì)胞IPO8蛋白相對(duì)表達(dá)量低于類似物陰性對(duì)照組(P0.05);轉(zhuǎn)染特異性miR-520b抑制物后,血管內(nèi)皮細(xì)胞中IPO8蛋白相對(duì)表達(dá)量高于抑制物對(duì)照組(P0.05)。提升miR-520b表達(dá)后人血管平滑肌細(xì)胞增殖和遷徙能力均低于相應(yīng)陰性對(duì)照(P0.05);抑制miR-520b表達(dá)后人血管平滑肌細(xì)胞增殖和遷徙能力均高于相應(yīng)陰性對(duì)照,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論動(dòng)脈粥樣硬化患者血清miR-520b表達(dá)水平明顯降低;miR-520b可通過阻斷NF-κB信號(hào)通路及抑制血管平滑肌細(xì)胞增殖、遷徙來發(fā)揮抗動(dòng)脈粥樣硬化的作用。
文內(nèi)圖片:miR-520b對(duì)人血管內(nèi)皮細(xì)胞中IPO8蛋白表達(dá)的影響
圖片說明:miR-520b對(duì)人血管內(nèi)皮細(xì)胞中IPO8蛋白表達(dá)的影響
[Abstract]:Objective to investigate the expression of serum miR-520b in patients with atherosclerosis and its related mechanism. Methods 42 patients with atherosclerosis were selected and 42 healthy elderly were collected as control. QRT-PCR was used to detect the expression of miR-520b in peripheral serum. Importin 8 (IPO8) was predicted by targetscan and miRanda database as the possible target gene of miR-520b, confirmed by double luciferase reporter gene detection, the effect of miR-520b on the expression of IPO8 in human vascular endothelial cells was detected by Western imprinting in vitro, and the effect of miR-520b on the proliferation and migration of human vascular smooth muscle cells was detected by CCK- 8 assay and Tranwells assay. Results the relative expression of serum miR-520b in patients with atherosclerosis was lower than that in healthy elderly (P 0.05). The detection of double luciferase reporter gene confirmed that IPO8 was the direct target gene of miR-520b; the relative expression of IPO8 protein in human vascular endothelial cells was lower than that in the control group (P 0.05), and the relative expression of IPO8 protein in vascular endothelial cells was higher than that in the inhibitor control group (P 0.05). The proliferation and migration ability of human vascular smooth muscle cells after increasing the expression of miR-520b was lower than that of the corresponding negative control (P 0.05), and the proliferation and migration ability of human vascular smooth muscle cells after inhibiting the expression of miR-520b was higher than that of the corresponding negative control, the difference was statistically significant (P 0.05). Conclusion the expression of miR-520b in serum of patients with atherosclerosis is significantly decreased, and miR-520b can play an anti-atherogenic role by blocking NF- kappa B signal pathway and inhibiting the proliferation and migration of vascular smooth muscle cells.
【作者單位】: 承德市中心醫(yī)院心血管內(nèi)科;
【基金】:河北省科學(xué)技術(shù)廳項(xiàng)目(20143217)
【分類號(hào)】:R543.5

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