Department of Critical Care Medicine, Yantaishan Hospital, Yantai 264001, China.
Department of Critical Care Medicine, Yantaishan Hospital, Yantai 264001, China.
Infect Genet Evol. 2021 Aug;92:104860. doi: 10.1016/j.meegid.2021.104860. Epub 2021 Apr 18.
To explore the role of lncRNA ZFAS1-mediated miR-193a-3p in the regulation of inflammatory responses in rats with sepsis-induced acute lung injury (ALI).
Sepsis-induced ALI models were constructed by LPS induction and then injected with ZFAS1 overexpression plasmid. Thereafter, lung injury score and the W/D weight ratio were calculated. Besides, bronchoalveolar lavage fluid (BALF) was isolated from rats to perform the cell count and protein quantification, while qRT-PCR and ELISA were performed to detect the inflammatory cytokines expressions. In vitro, NR8383 cells were transfected and then treated with LPS, followed by the measurement of inflammatory cytokines, cell viability and cell apoptosis.
In comparison with the Control group, rats in the LPS group presented sharp increases in the W/D weight ratio and injury score of lung, total protein concentration and the count of neutrophils and macrophages in BALF. Besides, rats in LPS group also resulted in a decrease in ZFAS1 expression and increase in miR-193a-3p expression in lung tissues, with the increased pro-inflammatory cytokines. Dual luciferase reporter gene assay confirmed a target relation between miR-193a-3p and ZFAS1. As compared to the Blank group, NR8383 cells in the LPS group had up-regulated pro-inflammatory cytokines with declined cell viability and elevated cell apoptosis; and meanwhile, ZFAS1 and Bcl-2 were decreased but miR-193a-3p and Bax were increased. Overexpression of ZFAS1 could significantly improve LPS-induced ALI in vivo and in vitro with reduced levels of pro-inflammatory cytokines.
Overexpression of ZFAS1, possibly via targeting the expression of miR-193a-3p, could inhibit the apoptosis and ameliorate the inflammatory responses of ALI in sepsis.
探讨长链非编码 RNA ZFAS1 介导的 miR-193a-3p 在脂多糖诱导的脓毒症大鼠急性肺损伤（ALI）中炎症反应调节中的作用。
通过 LPS 诱导构建脓毒症诱导的 ALI 模型，然后注射 ZFAS1 过表达质粒。此后，计算肺损伤评分和 W/D 重量比。此外，从大鼠中分离支气管肺泡灌洗液（BALF）进行细胞计数和蛋白定量，同时进行 qRT-PCR 和 ELISA 检测炎症细胞因子的表达。在体外，NR8383 细胞转染，然后用 LPS 处理，随后测量炎症细胞因子、细胞活力和细胞凋亡。
与对照组相比，LPS 组大鼠的 W/D 重量比和肺损伤评分、BALF 中的总蛋白浓度和中性粒细胞及巨噬细胞计数均明显升高。此外，LPS 组大鼠肺组织中 ZFAS1 表达降低，miR-193a-3p 表达增加，促炎细胞因子增加。双荧光素酶报告基因实验证实 miR-193a-3p 与 ZFAS1 之间存在靶关系。与空白组相比，LPS 组 NR8383 细胞中促炎细胞因子上调，细胞活力下降，细胞凋亡增加；同时，ZFAS1 和 Bcl-2 减少，而 miR-193a-3p 和 Bax 增加。过表达 ZFAS1 可显著改善 LPS 诱导的体内和体外 ALI，降低促炎细胞因子水平。
过表达 ZFAS1 可能通过靶向 miR-193a-3p 的表达抑制脓毒症中 ALI 的细胞凋亡并改善炎症反应。
