长链非编码 RNA 小核仁 RNA 宿主基因 1 通过调节 miR-181a-5p/XIAP 轴缓解脓毒症相关心肌损伤。
Long Non-Coding RNA Small Nucleolar RNA Host Gene 1 Alleviates Sepsis-Associated Myocardial Injury by Modulating the miR-181a-5p/XIAP Axis .
机构信息
Department of Emergency, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong Province, China
出版信息
Ann Clin Lab Sci. 2021 Mar;51(2):231-240.
OBJECTIVE
Sepsis is a systemic inflammatory response syndrome that results in severe myocardial injury. This study aimed to explore the role and mechanism of long non-coding RNA (lncRNA) small nucleolar RNA host gene 1 (SNHG1) in sepsis-induced myocardial injury .
METHODS
Embryonic rat ventricular myocardial cell line (H9c2) was treated with lipopolysaccharide (LPS) to simulate sepsis-induced myocardial injury . A quantitative real-time polymerase chain reaction was executed to determine the expression of SNHG1 and microRNA (miR)-181a-5p. 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-h-tetrazolium bromide assay was employed to measure cell viability. The levels of inflammatory factors (tumor necrosis factor alpha [TNF-α], interleukin 6 [IL-6], and IL-1β) were measured by enzyme-linked immunosorbent assay. Oxidative stress was assessed by measuring malondialdehyde, superoxide dismutase, and lactate dehydrogenase. The targeted interrelations among SNHG1, miR-181a-5p, and X-linked inhibitor of apoptosis protein (XIAP) were verified by dual-luciferase reporter assay. Relative protein expression of XIAP was detected by western blot.
RESULTS
SNHG1 and XIAP were down-regulated, and miR-181a-5p was up-regulated in LPS-induced H9c2 cells. Overexpression of SNHG1 or inhibition of miR-181a-5p facilitated cell viability and repressed inflammation and oxidative stress in LPS-treated H9c2 cells. MiR-181a-5p was a target of and negatively regulated by SNHG1. At the same time, XIAP was a target gene of and inversely modulated by miR-181a-5p. In addition, XIAP was positively regulated by SNHG1. Up-regulation of miR-181a-5p or silencing of XIAP reversed the inhibitory effects of SNHG1 on inflammation and oxidative stress, as well as the promoting effects on cell viability in LPS-induced H9c2 cells.
CONCLUSION
SNHG1 protected H9c2 cells against LPS-induced injury through modulating the miR-181a-5p/XIAP axis.
目的
脓毒症是一种全身性炎症反应综合征，可导致严重的心肌损伤。本研究旨在探讨长链非编码 RNA（lncRNA）小核仁 RNA 宿主基因 1（SNHG1）在脓毒症诱导的心肌损伤中的作用和机制。
方法
用脂多糖（LPS）处理胚胎大鼠心室心肌细胞系（H9c2）模拟脓毒症诱导的心肌损伤。采用实时定量聚合酶链反应测定 SNHG1 和 microRNA（miR）-181a-5p 的表达。3-（4,5-二甲基-2-噻唑基）-2,5-二苯基-2-H-四唑溴盐法测定细胞活力。酶联免疫吸附试验测定炎症因子（肿瘤坏死因子-α[TNF-α]、白细胞介素 6[IL-6]和白细胞介素 1β）水平。通过测定丙二醛、超氧化物歧化酶和乳酸脱氢酶评估氧化应激。通过双荧光素酶报告基因检测验证 SNHG1、miR-181a-5p 和 X 连锁凋亡抑制蛋白（XIAP）之间的靶向相互关系。通过 Western blot 检测 XIAP 的相对蛋白表达。
结果
LPS 诱导的 H9c2 细胞中 SNHG1 和 XIAP 下调，miR-181a-5p 上调。SNHG1 的过表达或 miR-181a-5p 的抑制促进 LPS 处理的 H9c2 细胞的细胞活力，并抑制炎症和氧化应激。miR-181a-5p 是 SNHG1 的靶基因，并对其负调控。同时，XIAP 是 miR-181a-5p 的靶基因，并对其反向调节。此外，SNHG1 正向调节 XIAP。上调 miR-181a-5p 或沉默 XIAP 逆转了 SNHG1 对 LPS 诱导的 H9c2 细胞炎症和氧化应激的抑制作用，以及对细胞活力的促进作用。
结论
SNHG1 通过调节 miR-181a-5p/XIAP 轴来保护 H9c2 细胞免受 LPS 诱导的损伤。
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