人肺動脈平滑肌細胞中微小RNA-210通過MKP-1負性調(diào)節(jié)低氧下細胞的增殖
本文關(guān)鍵詞:人肺動脈平滑肌細胞中微小RNA-210通過MKP-1負性調(diào)節(jié)低氧下細胞的增殖 出處:《中國病理生理雜志》2014年11期 論文類型:期刊論文
更多相關(guān)文章: 低氧性肺動脈高壓 人肺動脈平滑肌細胞 微小RNA- 絲裂原活化蛋白激酶磷酸酶-
【摘要】:目的:研究低氧條件下人肺動脈平滑肌細胞中微小RNA-210(miR-210)與絲裂原活化蛋白激酶磷酸酶1(MKP-1)的關(guān)系,是否通過MKP-1調(diào)節(jié)肺動脈平滑肌細胞增殖及其機制。方法:選用4~8代的人肺動脈平滑肌細胞,共分為12組,其中21%O2正常氧及1%O2低氧培養(yǎng)箱各6組,分別給予轉(zhuǎn)染miR-210抑制劑、增強劑、MKP-1 siRNA等處理,提取RNA和miRNA,并采用實時定量PCR法檢測各組平滑肌細胞中miR-210和MKP-1mRNA的表達,提取蛋白,并用Western blotting法比較各組MKP-1蛋白水平的表達,MTT法檢測肺動脈平滑肌細胞的增殖情況。結(jié)果:低氧下,人肺動脈平滑肌細胞中miR-210及MKP-1的表達均明顯增加,抑制miR-210表達使MKP-1的表達增加并可抑制低氧誘導的細胞增殖,miR-210的過表達可抑制低氧誘導的MKP-1表達上調(diào),但不影響細胞增殖,MKP-1基因沉默后,低氧下miR-210抑制劑對細胞增殖的抑制作用消失。結(jié)論:低氧下的人肺動脈平滑肌細胞中,MKP-1是miR-210的一個新的靶基因,MKP-1可以介導miR-210抑制劑對人肺動脈平滑肌細胞增殖的負性調(diào)節(jié)作用。
[Abstract]:Objective: To study the relationship between RNA-210 (miR-210) and mitogen activated protein kinase phosphatase 1 (MKP-1) in human pulmonary artery smooth muscle cells under hypoxia, and whether MKP-1 can regulate the proliferation of pulmonary artery smooth muscle cells and its mechanism. Methods: human pulmonary artery smooth muscle cells of 4~8 generation, were divided into 12 groups, including normal 21%O2 oxygen and 1%O2 hypoxia incubator for each of the 6 groups, were treated with miR-210 inhibitor, enhanced transfection agent, MKP-1 siRNA, RNA extraction and miRNA, and were detected by real-time quantitative PCR expression, miR-210 and MKP-1mRNA were smooth muscle cell protein extraction and Western blotting method to compare the expression levels of MKP-1 protein level, the detection of proliferation of pulmonary artery smooth muscle cells by MTT. Results: under hypoxia, the expression of miR-210 and MKP-1 in pulmonary artery smooth muscle cells were significantly increased, inhibiting the expression of miR-210 increased the expression of MKP-1 and inhibits hypoxia induced cell proliferation, overexpression of miR-210 can inhibit the expression of hypoxia induced MKP-1, but did not affect cell proliferation after MKP-1 gene silencing, inhibition effect of hypoxia on the proliferation of miR-210 inhibitors. Conclusion: MKP-1 is a new target gene of miR-210 in human pulmonary artery smooth muscle cells under hypoxia. MKP-1 can mediate the negative regulation effect of miR-210 inhibitors on the proliferation of human pulmonary artery smooth muscle cells.
【作者單位】: 山東大學附屬省立醫(yī)院兒科;
【基金】:山東省優(yōu)秀中青年科學家科研獎勵基金資助項目(No.BS 2011 SW 040)
【分類號】:R363
【正文快照】: 慢性低氧性肺動脈高壓是臨床許多心肺疾病發(fā)生發(fā)展過程中伴隨或最終的病理生理環(huán)節(jié),是一種嚴重的甚至危及生命的肺血管性疾病,臨床上表現(xiàn)為肺動脈壓力增高、肺血管阻力增加,右心室阻力負荷過重最終致右心衰竭,組織病理學改變?yōu)槔奂叭珜拥难苎?管壁的所有成分細胞均受累,最終
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