Emergency Department, the Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi Province, China.
Shock. 2023 Aug 1;60(2):206-213. doi: 10.1097/SHK.0000000000002169. Epub 2023 Aug 4.
Background: The dysregulation of circular RNAs (circRNAs) is involved in various human diseases, including sepsis-induced acute lung injury (ALI). We aimed to investigate the role of circTDRD9 in the development of sepsis-induced ALI. Methods: Cell models of sepsis-induced ALI were established by treating A549 cells with LPS. The expression of circTDRD9, miR-223-3p, and RAB10 mRNA was measured by quantitative real-time PCR (qPCR). The levels of inflammatory factors were measured by ELISA. Oxidative stress-related indicators were monitored by using commercial detection kits. The expression of fibrosis-related proteins was detected by Western blot assay. Cell proliferation was assessed by EdU assay. The predicted binding relationship between miR-223-3p and circTDRD9 or RAB10 was verified by dual-luciferase reporter assay, RIP assay or pull-down assay. Results: CircTDRD9 was highly expressed in LPS-treated A549 cells. CircTDRD9 downregulation prevented LPS-induced inflammation, oxidative stress, cell proliferation inhibition, and cell fibrosis in A549 cells, whereas these effects were reversed by the inhibition of miR-223-3p, a target of circTDRD9. In addition, RAB10 was verified as a target of miR-223-3p, and RAB10 overexpression recovered LPS-induced inflammation, oxidative stress, cell proliferation inhibition, and cell fibrosis in A549 cells that were ameliorated by miR-223-3p restoration. Importantly, circTDRD9 positively regulated RAB10 expression by binding to miR-223-3p. Conclusion: CircTDRD9 overexpression was closely associated with LPS-induced ALI. CircTDRD9 contributed to LPS-induced ALI partly by upregulating RAB10 via binding to miR-223-3p.
环状 RNA（circRNAs）的失调参与了多种人类疾病的发生，包括脓毒症诱导的急性肺损伤（ALI）。我们旨在研究 circTDRD9 在脓毒症诱导的 ALI 中的作用。
用 LPS 处理 A549 细胞建立脓毒症诱导的 ALI 细胞模型。通过定量实时 PCR（qPCR）测量 circTDRD9、miR-223-3p 和 RAB10 mRNA 的表达。通过 ELISA 测量炎症因子水平。通过使用商业检测试剂盒监测氧化应激相关指标。通过 Western blot 检测纤维化相关蛋白的表达。通过 EdU 检测评估细胞增殖。通过双荧光素酶报告基因实验、RIP 实验或下拉实验验证 miR-223-3p 与 circTDRD9 或 RAB10 的预测结合关系。
circTDRD9 在 LPS 处理的 A549 细胞中高表达。circTDRD9 下调可防止 LPS 诱导的 A549 细胞炎症、氧化应激、细胞增殖抑制和细胞纤维化，而 miR-223-3p 的抑制逆转了这些作用，miR-223-3p 是 circTDRD9 的靶标。此外，RAB10 被验证为 miR-223-3p 的靶标，RAB10 过表达恢复了 LPS 诱导的炎症、氧化应激、细胞增殖抑制和细胞纤维化，而 miR-223-3p 的恢复则减轻了这些作用。重要的是，circTDRD9 通过与 miR-223-3p 结合正向调节 RAB10 的表达。
circTDRD9 的过表达与 LPS 诱导的 ALI 密切相关。circTDRD9 通过与 miR-223-3p 结合上调 RAB10，从而导致 LPS 诱导的 ALI。
