MiR-200b-3p通过抑制HIF-1α表达减轻海水吸入诱导的肺水肿作用及机制  

miR-200b-3p suppressed the edema in seawater aspiration-induced ALI via inhibition the expression of HIF-1α

作　　者：张敏龙 杨翠平 王博[1] 崔云杰 金发光 Zhang Minlong;Yang Cuiping;Wang Bo;Cui Yunjie;Jin Faguang(Department of Respiratory and Critical Care Medicine,the 8th Medical Center,PLA General Hospital,Beijing 100091,China;Department of Respiratory,Tangdu Hospital,Air Force Military Medical University,Xi′an 710038,China;Department of Respiratory,Linxi Hospital,Chifeng 025250,China)

机构地区：[1]解放军总医院第八医学中心呼吸与危重症医学部,北京100091 [2]空军军医大学唐都医院呼吸科,西安710038 [3]内蒙古赤峰市林西县医院呼吸科,赤峰025250

出　　处：《中华肺部疾病杂志（电子版）》2024年第5期696-700,共5页Chinese Journal of Lung Diseases(Electronic Edition)

基　　金：解放军总医院第八医学中心国科金配套基金(QN202211004)。

摘　　要：目的分析miR-200b-3p在海水吸入性急性肺损伤(acute lung injury,ALI)中的作用及机制。方法选取32只健康SD大鼠,分为空白对照组、海水刺激组、miR-200b-3p antagomir预处理组和antag NC预处理组,每组8只,构建海水吸入性肺损伤大鼠及大鼠肺微血管内皮细胞(rat pulmonary microvascular endothelial cells,RPMVECs)模型。观察肺组织湿干比(W/D)、伊文思蓝法检测肺微血管通透性,Western blot检测低氧诱导因子-1(hypoxia inducible factor-1,HIF-1α)及血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达变化,采用双荧光素酶报告基因实验验证HIF-1α和miR-200b-3p的作用关系。结果海水干预4 h后,大鼠肺组织湿干比(W/D)及通透性明显增加,在肺组织及细胞模型中miR-200b-3p表达明显增高,HIF-1α表达出现升高,VEGF表达同时也出现上升。而抑制miR-200b-3p的表达后,肺水肿得到明显的减轻,肺组织及细胞模型中HIF-1α表达出现下降,VEGF表达也出现下降。双荧光素酶报告基因验证miR-200b-3p直接靶向作用于HIF-1α。与空白对照组对比,海水刺激组大鼠肺组织中HIF-1α及VEGF表达增加,RPMVECs细胞中HIF-1α及VEGF表达增加。结论增加表达的miR-200b-3p在海水吸入性肺损伤肺组织水肿的发展中起到关键作用,这种作用是通过调节HIF-1α/VEGF通路形成的。Objective To analyze the role and mechanism of miR-200b-3p in seawater inhalation acute lung injury(ALI).Methods Thirty-two healthy SD rats were selected and divided into blank control group,seawater stimulation group,miR-200b-3p antagomir pretreatment group and antag NC pretreatment group,with 8 rats in each group.The rat pulmonary microvascular endothelial cells(RPMVECs)model of seawater inhalation lung injury and rat pulmonary microvascular endothelial cells were constructed.Wet-dry ratio(W/D)of lung tissue was observed,and microvascular permeability of lung was detected by Evans blue method.Western blot analysis of the expression of-hypoxia inducible factor-1(HIF-1α)and vascular endothelial growth factor(VEGF)were detected.Dual luciferase reporter gene assay was used to verify the relationship between HIF-1αand miR-200b-3p.Results After seawater treatment for 4 h,the wet-dry ratio(W/D)and permeability of lung tissue were significantly increased,and the expression of miR-200b-3p,HIF-1αand VEGF were significantly increased in lung tissue and cell models.However,inhibition of the expression of miR-200b-3p significantly alleviated pulmonary edema,the expression of HIF-1αand VEGF were decreased in lung tissues and cell models.Dual luciferase reporter gene verified that miR-200b-3p directly targeted HIF-1α.Compared with blank control group,the expressions of HIF-1αand VEGF in lung tissue and HIF-1αand VEGF in RPMVECs cells were increased in seawater stimulated group.Conclusion Increased expression of miR-200b-3p plays a key role in the development of pulmonary edema induced by seawater aspiration injury,which is mediated by the HIF-1α/VEGF pathway.

关 键 词：急性肺损伤 海水 低氧诱导因子-1/血管内皮生长因子通路 miR-200b-3p 

分 类 号：R563[医药卫生—呼吸系统]