AIM To investigate the effect of miR-223 on NLRP3, subsequently regulating the production of the NLRP3/CASP1 inflammasome pathway-mediated proinflammatory cytokines IL-1β and IL-18 in human dental pulp fibroblasts (HDPFs). METHODOLOGY Human dental pulp tissue (HDPT) and HDPFs were obtained from impacted third molars. The miR-223 mimics and inhibitor or NLRP3 plasmid were used to upregulate or downregulate miR-223 or NLRP3 in HDPFs, respectively. Computational prediction via TargetScan 5.1 and a luciferase reporter assay was conducted to confirm target association. The mRNA and protein expression of NLRP3, caspase-1, IL-1β and IL-18 was determined by qRT-PCR and Western blotting, respectively. The release of IL-1β and IL-18 was analysed by ELISA. The significance of the differences between the experimental and the control groups was determined using one-way analysis of variance; P < 0.05 indicated statistical significance. RESULTS A decrease in miR-223 and an increase in NLRP3 in HDPT occurred during the transformation of reversible pulpitis into irreversible pulpitis compared to that in healthy pulp tissue (P < 0.05). The computational prediction and luciferase reporter assay confirmed that NLRP3 was a direct target of miR-223 in HDPFs. The miR-223 inhibitor further promoted ATP plus LPS-induced NLRP3/CASP1 inflammasome pathway activation compared to the ATP plus LPS-induced group (P < 0.05). In contrast, the miR-223 mimic significantly inhibited the NLRP3/CASP1 inflammasome pathway activation induced by ATP plus LPS compared to the ATP plus LPS-induced group (P < 0.05). CONCLUSION MiR-223 served as a negative regulator involved in the control of the production and secretion of proinflammatory cytokines mediated by the NLRP3/CASP1 inflammasome pathway by targeting NLRP3. These data provide insight into the potential regulatory effects of miRNAs on the NLRP3 inflammasome, thus opening up novel potential therapeutic avenues for future endodontic treatment.

中文翻译：

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MicroRNA-223 通过靶向人牙髓成纤维细胞中的 NLRP3 负调节 LPS 诱导的炎症反应。

目的 研究 miR-223 对 NLRP3 的影响，进而调节人牙髓成纤维细胞 (HDPFs) 中 NLRP3/CASP1 炎症小体通路介导的促炎细胞因子 IL-1β 和 IL-18 的产生。方法 人类牙髓组织 (HDPT) 和 HDPFs 从阻生的第三磨牙获得。miR-223 模拟物和抑制剂或 NLRP3 质粒分别用于上调或下调 HDPFs 中的 miR-223 或 NLRP3。通过 TargetScan 5.1 和荧光素酶报告基因分析进行计算预测以确认目标关联。分别通过qRT-PCR和Western印迹测定NLRP3、caspase-1、IL-1β和IL-18的mRNA和蛋白表达。通过ELISA分析IL-1β和IL-18的释放。使用单因素方差分析确定实验组和对照组之间差异的显着性；P < 0.05 表示有统计学意义。结果 与健康牙髓组织相比，可逆性牙髓炎向不可逆性牙髓炎转化过程中，HDPT中miR-223减少，NLRP3增加（P < 0.05）。计算预测和荧光素酶报告基因测定证实 NLRP3 是 miR-223 在 HDPFs 中的直接靶标。与 ATP 加 LPS 诱导组相比，miR-223 抑制剂进一步促进 ATP 加 LPS 诱导的 NLRP3/CASP1 炎性体通路激活（P < 0.05）。相比之下，与 ATP 加 LPS 诱导组相比，miR-223 模拟物显着抑制了 ATP 加 LPS 诱导的 NLRP3/CASP1 炎性体通路激活（P < 0.05）。结论 MiR-223 作为一种负调节剂，通过靶向 NLRP3 参与控制由 NLRP3/CASP1 炎性体通路介导的促炎细胞因子的产生和分泌。这些数据提供了对 miRNA 对 NLRP3 炎性体的潜在调节作用的深入了解，从而为未来的牙髓治疗开辟了新的潜在治疗途径。