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环状 RNA 0085289 通过调控 let-7f-5p/SOCS6 通路缓解牙周炎的进展。

Circ_0085289 Alleviates the Progression of Periodontitis by Regulating let-7f-5p/SOCS6 Pathway.

机构信息

Department of Stomatology, The Ninth People's Hospital of Chongqing, Chongqing, 400700, China.

Department of Oral Implantology, Stomatological Hospital of Chongqing Medical University, Chongqing, 401147, China.

出版信息

Inflammation. 2021 Aug;44(4):1607-1619. doi: 10.1007/s10753-021-01445-8. Epub 2021 Mar 12.

Abstract

Periodontitis is a common chronic inflammation that often occurs in adults. Circular RNAs (circRNAs) play a vital role in inflammation-related diseases. However, the role and potential basis of hsa_circ_0085289 in periodontitis remain unknown. Periodontal ligament cells (PDLCs) were exposed to lipopolysaccharide (LPS) to mimic periodontitis. The levels of circ_0085289, let-7f-5p, and suppressor of cytokine signaling 6 (SOCS6) were determined using qRT-PCR and western blot. The release of inflammatory cytokines was measured via enzyme-linked immunosorbent assay (ELISA). Cell viability and apoptosis were determined using Cell Counting Kit-8, flow cytometry, Caspase-3 Assay Kit, and western blot assays. The association between let-7f-5p and circ_0085289/SOCS6 was validated via dual-luciferase reporter, RNA pull-down, and RIP assays. Circ_0085289 and SOCS6 levels were reduced, and let-7f-5p level was increased in periodontitis patients and LPS-treated PDLCs. LPS stimulation caused PDLC injury and circ_0085289 downregulation. Moreover, circ_0085289 upregulation or let-7f-5p downregulation diminished LPS-triggered PDLC injury. Besides, circ_0085289 promoted SOCS6 expression by absorbing let-7f-5p. Circ_0085289 alleviated LPS-stimulated PDLC injury via targeting let-7f-5p. Moreover, let-7f-5p targeted SOCS6 to affect LPS-resulted PDLC injury. Circ_0085289 alleviated PDLC injury induced by LPS stimulation via modulating let-7f-5p/SOCS6 axis, suggesting a promising biomarker for periodontitis treatment.

摘要

牙周炎是一种常见的慢性炎症,常发生于成年人。环状 RNA(circRNA)在炎症相关疾病中发挥重要作用。然而,hsa_circ_0085289 在牙周炎中的作用及其潜在基础尚不清楚。用脂多糖(LPS)处理牙周韧带细胞(PDLCs)以模拟牙周炎。采用 qRT-PCR 和 Western blot 检测 circ_0085289、let-7f-5p 和细胞因子信号转导抑制因子 6(SOCS6)的水平。通过酶联免疫吸附试验(ELISA)测量炎症细胞因子的释放。采用细胞计数试剂盒-8(CCK-8)、流式细胞术、Caspase-3 测定试剂盒和 Western blot 检测细胞活力和凋亡。通过双荧光素酶报告、RNA 下拉和 RIP 测定验证 let-7f-5p 与 circ_0085289/SOCS6 的关联。牙周炎患者和 LPS 处理的 PDLCs 中 circ_0085289 和 SOCS6 水平降低,let-7f-5p 水平升高。LPS 刺激导致 PDLC 损伤和 circ_0085289 下调。此外,circ_0085289 上调或 let-7f-5p 下调可减轻 LPS 触发的 PDLC 损伤。此外,circ_0085289 通过吸收 let-7f-5p 促进 SOCS6 表达。circ_0085289 通过靶向 let-7f-5p 缓解 LPS 刺激的 PDLC 损伤。此外,let-7f-5p 靶向 SOCS6 影响 LPS 引起的 PDLC 损伤。circ_0085289 通过调节 let-7f-5p/SOCS6 轴缓解 LPS 刺激的 PDLC 损伤,提示其可能成为牙周炎治疗的有前途的生物标志物。

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