Bai Xiao-Zhi, Zhang Ju-Lei, Liu Yang, Zhang Wei, Li Xiao-Qiang, Wang Ke-Jia, Cao Meng-Yuan, Zhang Jia-Ning, Han Fu, Shi Ji-Hong, Hu Da-Hai
Department of Burns and Cutaneous Surgery, Xijing Hospital, the Fourth Military Medical University, Xi'an, China.
Department of Emergency, Xijing Hospital, the Fourth Military Medical University Xi'an, Xi'an, China.
Cell Physiol Biochem. 2018;49(2):489-500. doi: 10.1159/000492988. Epub 2018 Aug 29.
BACKGROUND/AIMS: With increased understanding of sepsis, mortality is decreasing. However, there is still a lack of effective therapeutic strategy. The inflammatory response of macrophages is critical during sepsis.
Macrophages were stimulated with LPS. Western blotting and qRT-PCR were used to detect inflammatory responses. Then, the inhibitor of microRNA-138 was transfected and Western blotting, qRT-PCR, H&E staining and ELISA were used to verify the role of microRNA-138 in inflammation. Then target gene prediction databases were used to predict the potential target of microRNA-138. Both animal and cell models under LPS challenges were established to verify the regulation of SIRT1 and microRNA-138 during inflammation.
The present study showed that microRNA-138 was increased in macrophages stimulated with LPS. Additionally, the NF-κB and AKT pathways were both activated. The pre-treatment of microRNA-138 inhibitor decreased inflammatory factors, downregulated the NF-κB pathway, activated the AKT pathway and protected against organ damage in mice challenged with LPS. SIRT1 was demonstrated as a potential target of microRNA-138In macrophages stimulated with LPS, the inhibition effect of microRNA-138 inhibitor on inflammation was lost by SIRT1 siRNA pre-treatment. In the animal model, the protective effect of microRNA-138 antagomir disappeared in SIRT1 knockout mice.
We demonstrated that miR-138 participated in the inflammatory process by inhibiting SIRT1 and activating the NF-κB pathway.
背景/目的：随着对脓毒症认识的增加，死亡率正在下降。然而，仍然缺乏有效的治疗策略。巨噬细胞的炎症反应在脓毒症期间至关重要。
用脂多糖刺激巨噬细胞。采用蛋白质免疫印迹法和定量逆转录-聚合酶链反应检测炎症反应。然后，转染微小RNA-138抑制剂，并使用蛋白质免疫印迹法、定量逆转录-聚合酶链反应、苏木精-伊红染色和酶联免疫吸附测定法验证微小RNA-138在炎症中的作用。然后使用靶基因预测数据库预测微小RNA-138的潜在靶标。建立脂多糖刺激下的动物和细胞模型，以验证炎症期间沉默信息调节因子1（SIRT1）和微小RNA-138的调控作用。
本研究表明，在脂多糖刺激的巨噬细胞中微小RNA-138增加。此外，核因子κB（NF-κB）和蛋白激酶B（AKT）信号通路均被激活。微小RNA-138抑制剂预处理可降低炎症因子水平，下调NF-κB信号通路，激活AKT信号通路，并保护脂多糖攻击的小鼠免受器官损伤。SIRT1被证明是微小RNA-138的潜在靶标。在用脂多糖刺激的巨噬细胞中，SIRT1小干扰RNA预处理消除了微小RNA-138抑制剂对炎症的抑制作用。在动物模型中，微小RNA-138拮抗剂的保护作用在SIRT1基因敲除小鼠中消失。
我们证明了微小RNA-138通过抑制SIRT1和激活NF-κB信号通路参与炎症过程。
