天津医药

天津医药 ›› 2019, Vol. 47 ›› Issue (8): 810-814.doi: 10.11958/20181913

• 实验研究 • 上一篇    下一篇

microRNA-125b在脓毒症急性肺损伤中的表达及其与炎症因子水平相关性

吴松林1，田小利2，徐陶1，雷贤英1，伍长学1，甘辞海1   

  1. 1四川泸州，西南医科大学附属医院重症医学科（邮编646000），2感染科
  • 收稿日期:2018-11-30 修回日期:2019-06-20 出版日期:2019-08-15 发布日期:2019-08-16
  • 通讯作者: 吴松林 E-mail:wslin919@163.com
  • 作者简介:吴松林（1984），男，硕士研究生，住院医师，主要从事脓毒血症、急性肺损伤方面研究
  • 基金资助:
    西南医科大学附属医院科研课题（microRNA-125b在脓毒症急性肺损伤中的表达及其免疫炎症调控机制研究）

Expression of microRNA-125b in acute lung injury induced by sepsis and its correlation with inflammatory factors

WU Song-lin1, TIAN Xiao-li2, XU Tao1, LEI Xian-ying1, WU Chang-xue1, GAN Ci-hai1   

  1. 1 Department of Critical Care Medicine, 2 Department of Infections, the Affiliated Hospital of Southwest Medical University, Luzhou 646000, China
  • Received:2018-11-30 Revised:2019-06-20 Published:2019-08-15 Online:2019-08-16
  • Contact: WU SONGLIN E-mail:wslin919@163.com
  • Supported by:
    Research on the research of microRNA-125b in acute lung injury of sepsis and its regulation mechanism of immune inflammation in the affiliated hospital of Southwest Medical University

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摘要： 摘要：目的探讨microRNA-125b（miR-125b）在脓毒症急性肺损伤中的表达及其与炎症因子的相关性。方法 腹腔注射脂多糖（LPS）诱导急性肺损伤大鼠模型，对照组注射等量的生理盐水（n=20）；各模型组（n=20）大鼠注射LPS 4、8、12及24 h后取其肺组织，HE染色观察病理学变化；PCR检测各组各时点肺组织中miR-125b、肿瘤坏死因子-α（TNF-α）与白细胞介素-6（IL-6）变化。Pearson相关性分析实验组肺组织中miR-125b的表达与TNF-α、IL-6水平的相关性。NR8383细胞培养后分组，对照组不进行LPS刺激，实验组加入LPS刺激4 h、8 h、12 h及24 h时收集细胞，检测各组miR-125b、TNF-α与IL-6的含量变化。另NR8383细胞设3组，阴性对照组只转染空表达质粒，miR-125b mimic组转染miR-125b mimic表达质粒，miR-125b inhibitor组转染miR-125b inhibitor质粒。PCR测各组细胞中miR-125b 表达情况，Western blot测Notch1的表达情况。结果LPS各处理组大鼠模型肺组织严重出血、水肿及大量炎症细胞浸润。与对照组比较，在注射LPS 4、8、12以24 h后，实验组大鼠肺组织中miR-125b显著降低；而肺组织中TNF-α与IL-6水平则升高，其中在LPS注射4 h后，TNF-α与IL-6的表达水平达到峰值（P＜0.05）。实验组肺组织中miR-125b的表达与TNF-α及IL-6呈负相关（r 分别为-0.599、-0.580；P＜0.05）。在LPS诱导的第4、8、12、24 h，实验组NR8383细胞系miR-125b表达水平均低于对照组，而TNF-α、IL-6的表达水平均高于对照组（P＜0.05）。与阴性对照组比较，miR-125b mimic组miR-125b 相对表达量升高、Notch1蛋白相对表达量则降低（P＜0.05），miR-125binhibitor组miR-125b 相对表达量降低、Notch1蛋白相对表达量升高（P＜0.05）；与miR-125b mimic组比较，miR-125binhibitor组miR-125b 相对表达量降低、Notch1蛋白相对表达量升高（P＜0.05）。结论在脓毒症急性肺损伤中，microRNA-125b可能通过调控Notch1蛋白的表达，影响炎症因子TNF-α与IL-6的表达，参与其免疫炎症调控过程。

关键词: 脓毒症, 急性肺损伤, 肿瘤坏死因子α, 白细胞介素6, miR-125b

Abstract: Objective To investigate the expression of microRNA125b in acute lung injury induced by sepsis, and its correlation with inflammatory factors. Methods The model rats of acute lung injury were induced by intraperitonealinjection of lipopolysaccharide (LPS) for 4, 8, 12 and 24 h (n=20). The control group was injected with the same amount of normal saline (n=20). The pathological changes of lung tissue were observed by HE staining. The changes of miR-125b,tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected by PCR in each group. Pearson correlation analysis was used to show the expression of miR-125b in lung tissue and its correlation with the levels of TNF-α and IL-6. NR8383 cells were cultured and subdivided into subgroups. The blank control group was not stimulated by LPS. The cells were collected after stimulation with LPS for 4 h, 8 h, 12 h and 24 h, and the levels of miR-125b, TNF-α and IL-6 were detected.NR8383 cells were set up into three groups, the negative control group was only transfected with the empty expression plasmid, and the miR-125b mimic group was transfected with the miR-125b mimic expression plasmid. The expression of miR-125b was detected by PCR in each group, and the expression of Notch1 was detected by Western blot assay. Results In the LPS treatment group, the lung tissue was severely hemorrhage, edema and a large amount of inflammatory cell after 24 hours of injection of LPS for 4, 8, and 12 hours in the experimental group, while the levels of TNF-α and IL-6 in thelung tissue were increased. The expression levels of TNF-α and IL-6 reached peak 4 h after LPS injection (P＜0.05).The expression of miR-125b in the lung tissue was negatively correlated with TNF-α and IL-6 in the experimental group (r=-0.599, -0.580, P＜0.05). The expression levels of miR-125b were lower in experiment group than those in the control group after LPS induction for 4 h, 8 h, 12 h and 24 h in the NR8383 cell line, while the expression levels of TNF-α and IL-6 were higher than those in the control group (P＜0.05). Compared with the negative control group, the relative expression level of miR-125b and the relative expression of Notch1 protein in miR-125b mimic group were decreased (P＜0.05), the relative expression of miR-125b in miR-125b inhibitor group was decreased, and the relative expression of Notch1 protein was decreased (P＜0.05). The amount of miR-125b decreased and the relative expression of Notch1 protein increased in miR-125b inhibitor group compared with those of miR-125b mimic group (P＜0.05). Conclusion In the acute lung injury induced by sepsis, microRNA-125b may regulate the expression of Notch1 protein, affect the expression of inflammatory factors TNF-α and IL-6, and participate in the process of immune inflammation regulation.

Key words: sepsis, acute lung injury, tumor necrosis factor-alpha, interleukin-6, microRNA-125b