LncRNA NEAT1 promotes inflammatory response in sepsis-induced liver injury via the Let-7a/TLR4 axis
LncRNA NEAT1 promotes inflammatory response in sepsis-induced liver injury via the Let-7a/TLR4 axis
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DOI：
10.1016/j.intimp.2019.105731
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发表时间：
2019-10-01
影响因子：
5.6
通讯作者：
Niu, Fang
中科院分区：
文献类型：
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作者：
Zhang, Cui-Cui;Niu, Fang
背景：脓毒症是一种全身性炎症反应，严重时可导致器官功能障碍和/或循环障碍。炎症反应失调在脓毒症引起的肝损伤中起着关键作用。多种microRNA和lncRNA已被证明参与炎症反应。方法：采用人肝组织和健康肝组织进行体内水平检测。采用Raw 264.7细胞和Kupffer细胞进行体外建模。分别采用q-PCR和酶联免疫吸附试验（ELISA）检测TNF-α、IL-6和IL-1 β的相对mRNA表达和蛋白水平。通过双荧光素酶报告基因分析检测lncRNA NEAT 1/Let-7a和Let-7a/TLR 4的结合。RNA免疫沉淀法（RIP）检测lncRNA NEAT 1与Let-7a的靶向性关系。结果：脓毒症肝损伤患者肝组织中存在lncRNA NEAT 1的过度表达，同时伴有Let-7a抑制和TLR 4激活。类似地，LPS刺激上调lncRNA NEAT 1表达，并且lncRNA NEAT 1抑制降低了体外炎性细胞因子的水平。Let-7a抑制剂处理以及TLR 4过表达挽救了lncRNA NEAT 1敲低细胞中炎性细胞因子的表达。结论：lncRNA NEAT 1与Let-7a相互作用，靶向TLR 4，参与LPS诱导的炎症反应。我们的检测方法可以为脓毒症诱导的肝损伤提供潜在的治疗靶点。
Background: Sepsis is a systemic inflammatory response that can lead to organ dysfunction and/or circulatory disorders in severe cases. The dysregulated inflammatory response plays a pivotal role in sepsis-induced liver injury. A variety of microRNAs and lncRNAs have been shown to be involved in the inflammatory response. However, their role in regulating sepsis-induced liver injury remains to be revealed.Methods: Human hepatic tissue and healthy tissue were used for in vivo level detection. And Raw264.7 cells and Kupffer cells were used for in vitro modelling. The relative mRNA expression and the protein levels of TNF-alpha, IL-6 and IL-1 beta were detected by q-PCR or by enzyme-linked immunosorbent assay (ELISA), respectively. The binding of lncRNA NEAT1/Let-7a and Let-7a/TLR4 was detected by dual-luciferase reporter assay. RNA Immunoprecipitation (RIP) was used to detect the targeting relationship between lncRNA NEAT1 and Let-7a. Western blotting (WB) was used to detect TLR4 expression in different cell models.Results: The overexpression of lncRNA NEAT1 accompanied by Let-7a inhibition and TLR4 activation was found in sepsis-induced liver injury patients. Similarly, LPS stimulation upregulated lncRNA NEAT1 expression, and lncRNA NEAT1 inhibition decreased the levels of inflammatory cytokines in vitro. Let-7a inhibitor treatment as well as TLR4 overexpression rescued the expression of inflammatory cytokines in lncRNA NEAT1-knockdown cells. Moreover, Let-7a interacted with both lncRNA NEAT1 and TLR4.Conclusion: We demonstrate that lncRNA NEAT1 interacts with Let-7a, targeting TLR4 to contribute to the LPS-induced inflammatory response. Our assay can provide a potential therapeutic target for sepsis-induced liver injury.