Circ_0138960敲低通过靶向牙髓炎中miR-545-5p/MYD88轴减轻脂多糖诱导的人牙髓细胞炎症反应和损伤。
Circ_0138960 knockdown alleviates lipopolysaccharide-induced inflammatory response and injury in human dental pulp cells by targeting miR-545-5p/MYD88 axis in pulpitis.
作者信息
Liang Changfu, Wu Wenzhi, He Xiaoning, Xue Fengming, Feng Daxing
机构信息
Department of Stomatology, The Second Affiliated Hospital of Hainan Medical University, Haikou 570100, China.
出版信息
J Dent Sci. 2023 Jan;18(1):191-202. doi: 10.1016/j.jds.2022.06.012. Epub 2022 Jul 2.
BACKGROUND/PURPOSE: Circular RNAs (circRNAs) have been shown to play important regulatory roles in many human diseases, yet their functions in pulpitis remain to be clarified. This study was designed to investigate the function of circ_0138960 in pulpitis progression and its underlying mechanism.
MATERIAL AND METHODS
Cell viability and proliferation were analyzed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 5-Ethynyl-2'-deoxyuridine (EdU) assay. Flow cytometry and enzyme-linked immunosorbent assay (ELISA) were conducted to analyze cell apoptosis rate and the release of inflammatory cytokines. The activity of superoxide dismutase (SOD) was analyzed using a SOD assay kit. Dual-luciferase reporter and RNA-pull down assays were performed to verify the interaction between microRNA-545-5p (miR-545-5p) and circ_0138960 or myeloid differentiation primary response gene 88 (MYD88).
RESULTS
Lipopolysaccharide (LPS) treatment restrained the proliferation and promoted the apoptosis, inflammation, and oxidative stress of human dental pulp cells (hDPCs). LPS treatment dose-dependently up-regulated circ_0138960 expression in hDPCs. Circ_0138960 knockdown overturned LPS-induced inflammation and injury in hDPCs. Circ_0138960 could act as a molecular sponge for miR-545-5p, and circ_0138960 knockdown protected hDPCs from LPS-induced effects by up-regulating miR-545-5p. miR-545-5p directly interacted with the 3' untranslated region (3'UTR) of MYD88, and MYD88 overexpression reversed miR-545-5p-mediated effects in LPS-treated hDPCs. Circ_0138960 positively regulated MYD88 expression by sponging miR-545-5p in hDPCs. LPS could activate nuclear factor kappa-B (NF-κB) signaling by targeting circ_0138960/miR-545-5p/MYD88 axis in hDPCs.
CONCLUSION
Circ_0138960 knockdown attenuated LPS-induced inflammatory response and injury in hDPCs by targeting the miR-545-5p/MYD88/NF-κB axis.
背景/目的：环状RNA（circRNAs）已被证明在多种人类疾病中发挥重要调节作用，但其在牙髓炎中的功能仍有待阐明。本研究旨在探讨circ_0138960在牙髓炎进展中的作用及其潜在机制。
材料与方法
采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法和5-乙炔基-2'-脱氧尿苷（EdU）法分析细胞活力和增殖情况。通过流式细胞术和酶联免疫吸附测定（ELISA）分析细胞凋亡率和炎性细胞因子的释放。使用超氧化物歧化酶（SOD）检测试剂盒分析SOD活性。进行双荧光素酶报告基因和RNA下拉实验，以验证微小RNA-545-5p（miR-545-5p）与circ_0138960或髓样分化初级反应基因88（MYD88）之间的相互作用。
结果
脂多糖（LPS）处理抑制了人牙髓细胞（hDPCs）的增殖，促进了其凋亡、炎症和氧化应激。LPS处理剂量依赖性地上调了hDPCs中circ_0138960的表达。敲低circ_0138960可逆转LPS诱导的hDPCs炎症和损伤。Circ_0138960可作为miR-545-5p的分子海绵，敲低circ_0138960通过上调miR-545-5p保护hDPCs免受LPS诱导的影响。miR-545-5p直接与MYD88的3'非翻译区（3'UTR）相互作用，过表达MYD88可逆转miR-545-5p在LPS处理的hDPCs中的介导作用。在hDPCs中，circ_0138960通过海绵吸附miR-545-5p正向调节MYD88的表达。LPS可通过靶向hDPCs中的circ_0138960/miR-545-5p/MYD88轴激活核因子κB（NF-κB）信号通路。
结论
敲低circ_0138960通过靶向miR-545-5p/MYD88/NF-κB轴减轻LPS诱导的hDPCs炎症反应和损伤。
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