Intensive Care Unit, Zhejiang Provincial People's Hospital, Hangzhou, Zhejiang Province, China.
Department of Infectious Diseases, Zhejiang Provincial People's Hospital, Hangzhou, Zhejiang Province, China.
Cell Biol Int. 2021 Aug;45(8):1666-1675. doi: 10.1002/cbin.11601. Epub 2021 Apr 9.
Sepsis-induced acute lung injury is associated with dysregulated inflammatory reactions. MiR-19b-3p level was reported to be downregulated in patients with sepsis. To evaluate the role of miR-19b-3p in sepsis, cecum ligation and puncture-induced mouse sepsis model and lpopolysaccharide (LPS)-treated pulmonary microvascular endothelial cells (PMVECs) were used. For in vivo study, lung tissue was harvested for hematoxylin and eosin (H&E) staining, tumor necrosis factor-α, interleukin-6 (IL-6), IL-1β, and p-p65, p-IκB measuring. Cell apoptosis was assessed by TUNEL assay. For in vitro study, cell proliferation and apoptosis were detected by CCK-8 and flow cytometry, respectively. Methylation of miR-19b-3p promoter was measured by methylation-specific PCR (MSP) assay. The target of miR-19b-3p was determined by dual-luciferase reporter gene assay. The level of miR-19b-3p was determined to be downregulated in vitro and in vivo. In addition, miR-19b-3p protected mice from inflammation injury through inhibiting NF-κB signaling pathway. Overexpression of miR-19b-3p increased cell viability, decreased apoptosis, and proinflammatory cytokines secretion in LPS-treated PMVECs. Besides these, Krüppel-like factor 7 (KLF7) was confirmed as the target of miR-19b-3p. And methylation of miR-19b-3p was the reason of decreased miR-19b-3p level. In conclusion, miR-19b-3p protected cells from sepsis-induced inflammation injury via inhibiting NF-κB signaling pathway, and KLF7 was a potential target.
脓毒症诱导的急性肺损伤与失调的炎症反应有关。有报道称，脓毒症患者的 miR-19b-3p 水平下调。为了评估 miR-19b-3p 在脓毒症中的作用，使用盲肠结扎和穿刺诱导的小鼠脓毒症模型和脂多糖（LPS）处理的肺微血管内皮细胞（PMVEC）进行研究。在体内研究中，采集肺组织进行苏木精和伊红（H&E）染色、肿瘤坏死因子-α、白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）和 p-p65、p-IκB 测量。通过 TUNEL 测定评估细胞凋亡。在体外研究中，通过 CCK-8 和流式细胞术分别检测细胞增殖和凋亡。通过甲基化特异性 PCR（MSP）测定测量 miR-19b-3p 启动子的甲基化。通过双荧光素酶报告基因测定确定 miR-19b-3p 的靶标。结果显示，miR-19b-3p 的水平在体外和体内均下调。此外，miR-19b-3p 通过抑制 NF-κB 信号通路保护小鼠免受炎症损伤。过表达 miR-19b-3p 增加了 LPS 处理的 PMVECs 中的细胞活力，减少了细胞凋亡和促炎细胞因子的分泌。除此之外，Krüppel 样因子 7（KLF7）被确认为 miR-19b-3p 的靶标。并且 miR-19b-3p 的甲基化是 miR-19b-3p 水平降低的原因。综上所述，miR-19b-3p 通过抑制 NF-κB 信号通路保护细胞免受脓毒症诱导的炎症损伤，而 KLF7 是一个潜在的靶标。
