Mo Song, Yi Qushen, Bei Xuezhu, Huang Yuan, Lai Junhua
Department of Intensive Care Unit, Liuzhou Worker's Hospital, The Fourth Affiliated Hospital of Guangxi Medical University, Liuzhou, China.
Shock. 2023 Oct 1;60(4):565-572. doi: 10.1097/SHK.0000000000002196.
Background: Sepsis-associated acute lung injury (SA-ALI) is a serious threat to human health. A growing body of evidence suggested that circular RNAs may be involved in ALI progression. The aim of this study was to investigate the effect and mechanism of circ_0001226 on lipopolysaccharide (LPS)-induced BEAS-2B cells. Methods: BEAS-2B cells were stimulated with LPS to establish a SA-ALI cell model. The expression of circ_0001226, miR-940, and transforming growth factor beta receptor II (TGFBR2) were monitored by quantitative real-time polymerase chain reaction. Cell proliferation and apoptosis were evaluated by the Cell Counting Kit-8, 5-ethynyl-2'-deoxyuridine assay, and flow cytometry. The levels of interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-α were calculated by enzyme-linked immunosorbent assay. Western blot was implemented to test the protein levels of PCNA, Bax, and TGFBR2. Dual-luciferase reporter assay and RNA pull-down assay were adopted to investigate the interaction between circ_0001226 and miR-940, as well as TGFBR2 and miR-940. Results: The levels of circ_0001226 and TGFBR2 were elevated, and miR-940 was decreased in SA-ALI serum specimens and LPS-evoked BEAS-2B cells. Besides that, circ_0001226 interference contributed to cell proliferation and restrained apoptosis and inflammation in LPS-induced BEAS-2B cells. Mechanically, circ_0001226 worked as a molecular sponge of miR-940 to regulate TGFBR2 expression. Conclusion: Circ_0001226 deficiency weakened LPS-mediated proliferation inhibition and inflammatory processes in BEAS-2B cells by binding miR-940 and regulating TGFBR2.
脓毒症相关急性肺损伤（SA - ALI）对人类健康构成严重威胁。越来越多的证据表明，环状RNA可能参与ALI的进展。本研究旨在探讨circ_0001226对脂多糖（LPS）诱导的BEAS - 2B细胞的影响及其机制。方法：用LPS刺激BEAS - 2B细胞以建立SA - ALI细胞模型。通过定量实时聚合酶链反应监测circ_0001226、miR - 940和转化生长因子β受体II（TGFBR2）的表达。采用细胞计数试剂盒 - 8、5 - 乙炔基 - 2'-脱氧尿苷测定法和流式细胞术评估细胞增殖和凋亡。通过酶联免疫吸附测定法计算白细胞介素 - 6（IL - 6）、IL - 1β和肿瘤坏死因子 - α的水平。采用蛋白质免疫印迹法检测增殖细胞核抗原（PCNA）、Bax和TGFBR2的蛋白水平。采用双荧光素酶报告基因测定法和RNA下拉测定法研究circ_0001226与miR - 940以及TGFBR2与miR - 940之间的相互作用。结果：在SA - ALI血清标本和LPS诱导的BEAS - 2B细胞中，circ_0001226和TGFBR2水平升高，而miR - 940水平降低。此外，circ_0001226干扰促进了LPS诱导的BEAS - 2B细胞的增殖，并抑制了其凋亡和炎症。机制上，circ_0001226作为miR - 940的分子海绵来调节TGFBR2的表达。结论：Circ_0001226缺陷通过结合miR - 940并调节TGFBR2减弱了LPS介导的BEAS - 2B细胞增殖抑制和炎症过程。
