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50ng/ml佛波酯(PMA)诱导成熟的U937细胞随机分为对照组和LPS(10μg/ml)刺激组,LPS刺激时间为1、3、6h,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^(-1)β)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^(-1)βmRNA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-αmRNA表达量(r=0.835,P<0.01)、IL^(-1)βmRNA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。
Objective To study the expression of inflammatory factors reregulated by microRNA-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（10μg/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-αand IL-1βin 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-αmRNA（r = 0.835,P 〈0.01）and IL-1βmRNA（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.
作者
唐佳雯
张颖
纪健
娄晓丽
朱涛
TANG Jiawen;ZHANG Ying;JI Jian;LOU Xiaoli;ZHU Tao(Nanjing Medical University,Nanjing 210029,China)
出处
《临床麻醉学杂志》
CAS
CSCD
北大核心
2018年第8期800-804,共5页
Journal of Clinical Anesthesiology
基金
上海市科委医学引导项目基金(134119b2700)
