下调长链非编码 RNA LINC00472 通过调节 miR-373-3p/TRIM8 轴缓解脓毒症诱导的急性肝损伤。
Down-regulation of long noncoding RNA LINC00472 alleviates sepsis-induced acute hepatic injury by regulating miR-373-3p/TRIM8 axis.
机构信息
Department of Pediatrics, Jinan Maternity and Child Care Hospital, Jinan 250001, China.
Department of Pediatrics, Jinan Central Hospital, Jinan 250021, China.
出版信息
Exp Mol Pathol. 2020 Dec;117:104562. doi: 10.1016/j.yexmp.2020.104562. Epub 2020 Oct 28.
BACKGROUND
The long noncoding RNAs (lncRNAs) have been confirmed to be involved in sepsis-induced organ injury. Here, we first investigated the functional role and the underlying mechanism of lncRNA LINC00472 in sepsis-induced acute hepatic injury (AHI).
METHODS
Human liver THLE-3 cells were treated with lipopolysaccharide (LPS) to mimic sepsis-induced AHI in vitro; intraperitoneal injection of LPS in rats were used as an in vivo model of AHI induced by sepsis. The expressions of LINC00472, miR-373-3p, and TRIM8 mRNA were detected by qRT-PCR. The effects of LINC00472 and miR-373-3p on the viability of THLE-3 cells were assessed by CCK-8 assay. Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays were performed to determine the binding relationship between LINC00472 and miR-373-3p as well as between miR-373-3p and TRIM8. The expressions of apoptosis-related proteins and TRIM8 were detected by Western blot; the levels of ALT, AST, TNF-α, IL-6, and IL-10 in the serum of rats were measured using ELSA assay.
RESULTS
LINC00472 and TRIM8 were significantly upregulated in liver tissues and THLE-3 cells in sepsis-induced AHI models, while miR-373-3p was downregulated. Silencing of LINC00472 promoted cell viability and suppressed cell apoptosis in LPS-treated THLE-3 cells, whereas upregulation of LINC00472 had the opposite effect. Moreover, LINC00472 served as a sponge for miR-373-3p and negatively regulated its expression. miR-373-3p mimics could promote THLE-3 cell viability and suppress cell apoptosis. Additionally, TRIM8 was a direct target of miR-373-3p, which was downregulated in LINC00472-silenced cells and upregulated by the miR-373-3p inhibitor. Further, the co-transfection of miR-373-3p inhibitor reversed the effects of LINC00472 knockdown on cell viability and apoptosis. Downregulation of LINC00472 in rats restored the levels of ALT, AST, IL-6, IL-10, and TNF-α.
CONCLUSION
Downregulation of LINC00472 ameliorates sepsis-induced AHI by regulating the miR-373-3p/TRIM8 axis.
背景
长链非编码 RNA（lncRNA）已被证实参与了脓毒症引起的器官损伤。在这里，我们首次研究了 lncRNA LINC00472 在脓毒症诱导的急性肝损伤（AHI）中的功能作用和潜在机制。
方法
用脂多糖（LPS）处理人肝 THLE-3 细胞，在体外模拟脓毒症诱导的 AHI；用腹腔注射 LPS 的大鼠作为脓毒症诱导的 AHI 的体内模型。通过 qRT-PCR 检测 LINC00472、miR-373-3p 和 TRIM8 mRNA 的表达。通过 CCK-8 测定评估 LINC00472 和 miR-373-3p 对 THLE-3 细胞活力的影响。通过双荧光素酶报告和 RNA 免疫沉淀（RIP）测定确定 LINC00472 与 miR-373-3p 以及 miR-373-3p 与 TRIM8 之间的结合关系。通过 Western blot 检测凋亡相关蛋白和 TRIM8 的表达；通过 ELISA 测定大鼠血清中 ALT、AST、TNF-α、IL-6 和 IL-10 的水平。
结果
在脓毒症诱导的 AHI 模型中，LINC00472 和 TRIM8 在肝组织和 THLE-3 细胞中均显著上调，而 miR-373-3p 下调。沉默 LINC00472 可促进 LPS 处理的 THLE-3 细胞的活力并抑制细胞凋亡，而上调 LINC00472 则有相反的效果。此外，LINC00472 可作为 miR-373-3p 的海绵，并负调控其表达。miR-373-3p 模拟物可促进 THLE-3 细胞活力并抑制细胞凋亡。此外，TRIM8 是 miR-373-3p 的直接靶标，在 LINC00472 沉默的细胞中下调，并被 miR-373-3p 抑制剂上调。进一步，miR-373-3p 抑制剂的共转染逆转了 LINC00472 敲低对细胞活力和凋亡的影响。在大鼠中下调 LINC00472 可恢复 ALT、AST、IL-6、IL-10 和 TNF-α 的水平。
结论
下调 LINC00472 通过调节 miR-373-3p/TRIM8 轴改善脓毒症引起的 AHI。
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