Zhang Pingli, Huang Qikun, Liao Peiyuan, Sun Shijing, Yang Jing
Department of Pediatric, Qilu Hospital (Qingdao), Cheeloo College of Medicine, Shandong University Qingdao 266035, Shandong, China.
Am J Transl Res. 2023 Jun 15;15(6):3900-3911. eCollection 2023.
MicroRNA-23a-3p (miR-23a) is a novel gene regulator involved in inflammation. This study aimed to explore the molecular mechanism of miR-23a in sepsis-induced lung injury both and .
Lipopolysaccharide (LPS)- and ATP-stimulated human myeloid leukemia mononuclear cells (THP-1) and Human Bronchial Epithelial Cells (BEAS-2B) cell lines were used, while cecal ligation and puncture (CLP)-induced sepsis BABL/c mice were constructed. The mRNA expression levels of interleukin (IL)-18, IL-1β, and miR-23a were determined, and Western blotting was used to measure CXCR4/PTEN/PI3K/AKT signaling. The concentrations of cytokines and Nod-like receptor family pyrin domain-containing 3 (NLRP3) were determined using an enzyme-linked immunosorbent assay. Lung tissue of mice was subjected to hematoxylin-eosin staining for examining myocardial injury.
MiR-23a inhibited NLRP3 inflammasome activation in LPS- and ATP-stimulated THP-1 and BEAS-2B cells (<0.05). Overexpression of miR-23a decreased the lactate dehydrogenase release rate in the cells (<0.05). Meanwhile, miR-23a overexpression decreased the concentration and gene expression of IL-1β and IL-18 in CXCR4 positive cells (<0.05). Conversely, miR-23a knockdown increased the concentration and gene expression of IL-1β and IL-18 (<0.05). Additionally, PTEN and p53 proteins were up-regulated in miR-23a mimic group and down-regulated in miR-23a inhibitor group (<0.05). Furthermore, miR-23a expression was decreased in sepsis-induced lung injury mice (<0.05). MiR-23a overexpression reduced the sepsis-induced lung injury probably by inhibiting acetylcholinesterase activity and expression levels of IL-1β, IL-18, capase-1, and NLRP3 (<0.05).
miR-23a can significantly alleviate sepsis-induced lung injury in CLP-induced septic mice and LPS-stimulated cell lines by suppressing NLRP3 inflammasome activation and inflammatory response, while promoting the CXCR4/PTEN/PI3K/AKT pathway.
微小RNA-23a-3p（miR-23a）是一种参与炎症反应的新型基因调节因子。本研究旨在探究miR-23a在脓毒症诱导的肺损伤中的分子机制。
使用脂多糖（LPS）和ATP刺激的人髓系白血病单核细胞（THP-1）和人支气管上皮细胞（BEAS-2B）细胞系，同时构建盲肠结扎和穿刺（CLP）诱导的脓毒症BABL/c小鼠模型。测定白细胞介素（IL）-18、IL-1β和miR-23a的mRNA表达水平，并采用蛋白质免疫印迹法检测CXCR4/PTEN/PI3K/AKT信号通路。使用酶联免疫吸附测定法测定细胞因子和含NOD样受体家族吡咯结构域蛋白3（NLRP3）的浓度。对小鼠肺组织进行苏木精-伊红染色以检查心肌损伤。
miR-23a抑制LPS和ATP刺激的THP-1和BEAS-2B细胞中NLRP3炎性小体的激活（<0.05）。miR-23a过表达降低了细胞中乳酸脱氢酶的释放率（<0.05）。同时，miR-23a过表达降低了CXCR4阳性细胞中IL-1β和IL-18的浓度及基因表达（<0.05）。相反，miR-23a敲低增加了IL-1β和IL-18的浓度及基因表达（<0.05）。此外，PTEN和p53蛋白在miR-23a模拟物组中上调，在miR-23a抑制剂组中下调（<0.05）。此外，脓毒症诱导的肺损伤小鼠中miR-23a表达降低（<0.05）。miR-23a过表达可能通过抑制乙酰胆碱酯酶活性以及IL-1β、IL-18、半胱天冬酶-1和NLRP3的表达水平减轻脓毒症诱导的肺损伤（<0.05）。
miR-23a可通过抑制NLRP3炎性小体激活和炎症反应，同时促进CXCR4/PTEN/PI3K/AKT通路，显著减轻CLP诱导的脓毒症小鼠和LPS刺激的细胞系中脓毒症诱导的肺损伤。
