miR-199a-5p抑制低氧条件下大鼠气道平滑肌增殖和低氧诱导因子1α表达

发布时间：2018-04-06 07:31

本文选题：miR-a-p　切入点：低氧诱导因子α　出处：《细胞与分子免疫学杂志》2015年09期

【摘要】：目的探讨miR-199a-5p在常氧及低氧环境下的大鼠气道平滑肌细胞(ASMC)中的表达以及miR-199a-5p对ASMC增殖的调控和低氧诱导因子1α(HIF-1α)表达的影响。方法用贴壁法体外培养大鼠ASMC,在常氧及低氧条件下培养24 h,实时荧光定量PCR(qRT-PCR)检测ASMC中miR-199a-5p含量;人工合成大鼠miR-199a-5p的拟似物和抑制物,用脂质体转入低氧培养的ASMC,qRT-PCR检测miR-199a-5p及HIF-1αmRNA的表达,Western blot法检测HIF-1α蛋白的表达,CCK-8法检测ASMC的增殖情况。结果与常氧组相比,低氧处理促进细胞增殖,低氧组HIF-1αmRNA及HIF-1α蛋白的相对水平显著升高;低氧组miR-199a-5p的量较常氧组减少;转染miR-199a-5p拟似物可以显著减缓低氧条件下ASMC的增殖,而其抑制物可促进低氧条件下的细胞增殖。拟似物组HIF-1α蛋白较对照组降低,抑制物组则升高,而低氧条件下各组HIF-1αmRNA水平无明显差异。结论 miR-199a-5p能抑制低氧条件下ASMC的增殖和HIF-1α蛋白的表达。
[Abstract]:Objective to investigate the effect of miR-199a-5p in normoxia and hypoxia condition of rat airway smooth muscle cells (ASMC) in the expression and regulation of miR-199a-5p on the proliferation of ASMC and hypoxia inducible factor 1 alpha (HIF-1 alpha) was studied. Methods using adherent method in vitro cultured rat ASMC under normoxia and hypoxia cultured under 24 h, real time fluorescent quantitative PCR (qRT-PCR) to detect the content of miR-199a-5p in ASMC; analogue and synthetic inhibitor miR-199a-5p in rats, using liposome into hypoxia ASMC, miR-199a-5p and HIF-1 to detect the expression of qRT-PCR alpha mRNA, expression of Western blot was detected by HIF-1 protein and proliferation of ASMC CCK-8 detection method. Results compared with normoxia group, hypoxia can promote cell proliferation and relative levels of hypoxia group HIF-1 alpha mRNA and HIF-1 alpha protein was significantly increased; the amount of miR-199a-5p in hypoxia group compared with normoxia group decreased; transfection of miR-199a-5p analogues can significantly Slow under hypoxic conditions ASMC proliferation, and its inhibitor can promote cell proliferation under hypoxic conditions. Analogue of HIF-1 alpha protein group is lower than control group, inhibitor group was increased, while no significant difference under hypoxic conditions were HIF-1 alpha level of mRNA. Conclusion: miR-199a-5p can inhibit the expression of HIF-1 protein and proliferation of alpha ASMC hypoxia a.

【作者单位】：徐州医学院附属医院呼吸内科;
【分类号】：R562.25

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