LncRNA-5657 silencing alleviates sepsis-induced lung injury by suppressing the expression of spinster homology protein 2
LncRNA-5657 silencing alleviates sepsis-induced lung injury by suppressing the expression of spinster homology protein 2
复制标题
LncRNA-5657 沉默通过抑制老处女同源蛋白 2 的表达来减轻脓毒症引起的肺损伤。
DOI：
10.1016/j.intimp.2020.106875
复制
发表时间：
2020-11-01
影响因子：
5.6
通讯作者：
Qian, Kejian
中科院分区：
文献类型：
--
作者：
Liu, Fen;Hu, Shilin;Qian, Kejian
背景：LncRNAs与人类许多重大疾病密切相关，但LncRNAs在脓毒症诱导的肺损伤中的作用尚不清楚。方法：采用高通量测序检测LPS刺激后肺泡巨噬细胞lncRNA表达谱的变化。收集脓毒症致肺损伤患者的BALF。用sh-lncRNA-5657静脉注射治疗clp诱导的脓毒性肺损伤大鼠。用表达lncRNA-5657、lncRNA-5657智能消音器或si-spns2的慢病毒载体转染NR8383细胞。检测患者、大鼠BALF及巨噬细胞中lncRNA-5657及促炎细胞因子的表达水平。观察各组组织病理学评分、肺干湿比及巨噬细胞中spns2表达的变化。lncRNA-5657与其潜在靶基因spns2之间的关系通过双荧光素酶报告基因实验得到验证。结果：lps刺激的巨噬细胞lncRNA表达谱显示，lncRNA-5657的折叠变化最大。脓毒症致ARDS患者的BALF细胞和clp致脓毒症大鼠肺组织中lncRNA-5657水平显著升高。LncRNA-5657沉默可减轻clp诱导的大鼠肺部炎症。在NR8383细胞中，lncRNA-5657过表达增强，而lncRNA-5657沉默则减弱促炎细胞因子和spns2的表达。双荧光素酶报告基因分析显示，lncRNA-5657与spns2基因的启动子相互作用。Spns2沉默可减轻lps诱导的炎症反应，阻断lncRNA-5657在肺泡巨噬细胞中的促炎功能。结论：LncRNA-5657与败血症所致肺损伤密切相关。体外和体内数据表明，LncRNA-5657沉默通过抑制spns2表达抑制肺部炎症反应，减轻败血症诱导的肺损伤。
Background: LncRNAs are closely associated with many human major diseases, however, the roles of lncRNAs in sepsis-induced lung injury remain unclear.Methods: High-throughput sequencing was used to detect the changes in lncRNA expression profile in alveolar macrophages after LPS stimulation. The BALF of patients with sepsis-induced lung injury was collected. Rats with CLP-induced septic lung injury were treated with sh-lncRNA-5657 via intravenous injection. NR8383 cells were transfected with lentiviral vectors expressing lncRNA-5657, lncRNA-5657 smart silencer, or si-spns2. The BALF cells expression levels of lncRNA-5657 and proinflammatory cytokines in BALF of patients and rats as well as in rat macrophages were measured. Changes in histopathologic score, lung wet/dry weight ratio, and spns2 expression in macrophages were examined. The relationship between lncRNA-5657 and its potential target gene spns2 was validated using a dual-luciferase reporter assay.Results: The lncRNA expression profile of LPS-stimulated macrophages demonstrated that lncRNA-5657 showed the greatest fold-change. The BALF cells of patients with sepsis-induced ARDS and the lung tissue of rats with CLP-induced sepsis had significantly increased lncRNA-5657 levels. LncRNA-5657 silencing alleviated CLP-induced lung inflammation in rats. In NR8383 cells, lncRNA-5657 overexpression enhanced, whereas lncRNA-5657 silencing attenuated the expression of proinflammatory cytokines and spns2. A dual-luciferase reporter assay showed that lncRNA-5657 interacted with the promoter of the spns2 gene. Spns2 silencing alleviated LPS-induced inflammatory response and blocked the proinflammatory function of lncRNA-5657 in alveolar macrophages.Conclusion: LncRNA-5657 is closely associated with sepsis-induce lung injury. In vitro and in vivo data demonstrated that LncRNA-5657 silencing alleviates sepsis-induced lung injury by inhibiting lung inflammatory response via suppressing spns2 expression.