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脂多糖对U937细胞中microRNA-107与NF-κB信号通路的影响 |
Effect of lipopolysaccharide on the expression of microRNA-107 and NF-κB signaling in U937 cells |
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DOI:10.12089/jca.2018.08.017 |
中文关键词: microRNA-107 核因子-κB 脂多糖 U937细胞 |
英文关键词: MicroRNA-107 Nuclear factor-κB Lipopolysaccharide U937 macrophages |
基金项目:上海市科委医学引导项目基金(134119b2700) |
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中文摘要: |
目的 分析单核细胞在炎症状态下microRNA-107与NF-κB信号通路的相关性。
方法 经50 ng/ml佛波酯(PMA)诱导成熟的U937细胞随机分为对照组和LPS(10 μg/ml)刺激组,LPS刺激时间为1、3、6 h,采用qRT-PCR检测各组细胞中microRNA-107、肿瘤坏死因子α(TNF-α)、白细胞介素-1β(IL-1β)的mRNA表达量;Western blot法检测细胞NF-κB(p65)与磷酸化p65(p-p65)蛋白含量。
结果 与对照组比较,LPS组细胞microRNA-107、TNF-α、IL-1β mRNA表达量明显升高(P<0.05);p-p65/p65蛋白含量比值明显升高(P<0.01)。Spearman相关性分析显示,microRNA-107表达量与p-p65/p65蛋白含量比值呈中度正相关(r=0.69,P<0.01);与TNF-α mRNA表达量(r=0.835,P<0.01)、IL-1β mRNA表达量(r=0.839,P<0.01)呈高度正相关。
结论 脂多糖刺激PMA诱导的U937细胞后microRNA-107的表达升高可能与NF-κB信号通路有关。 |
英文摘要: |
Objective To study the expression of inflammatory factors reregulated by micro-RNA-107 and NF-κB signaling pathways.
Methods An in vitro monocyte inflammatory model in PMA-induced U937 cells was established by administration of lipopolysaccharide (LPS).The differentiation of U937 cells was induced by phorbol ester (50 ng/ml) for 1 hour, 3 hours and 6 hours. Cells were treated with either PBS or LPS (10 μg/ml) at each time point.The levels of microRNA-107, tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were detected by qRT-PCR.Western blot was carried out to investigate the expression levels of NF-κB (p65) and phosphorylated-p65 (p-p65) protein.
Results Compared with the control group, the mRNA levels of microRNA-107, TNF-α and IL-1β in LPS-stimulated cells were significantly increased (P < 0.05). Also the protein expression of p-p65/p65 was higher in the control group (P < 0.01). Spearman correlation analysis showed that a positive linear correlation between microRNA-107 and the expression of p-p65/p65 protein (r = 0.69, P < 0.01). MicroRNA-107 was positively correlated with TNF-α mRNA (r = 0.835, P < 0.01) and IL-1β mRNA (r = 0.839, P < 0.01).
Conclusion The increased expression of microRNA-107 in PMA-induced U937 cells may be related to the NF-κB signaling pathway after LPS stimulation. |
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