MicroRNA-223 negatively regulates LPS-induced inflammatory responses by targeting NLRP3 in human dental pulp fibroblasts
MicroRNA-223 negatively regulates LPS-induced inflammatory responses by targeting NLRP3 in human dental pulp fibroblasts
复制标题
MicroRNA-223 通过靶向人牙髓成纤维细胞中的 NLRP3 负向调节 LPS 诱导的炎症反应
DOI：
10.1111/iej.13413
复制
发表时间：
2020-10-17
影响因子：
5
通讯作者：
Jiang, W.
中科院分区：
文献类型：
--
作者：
Wang, D.;Sun, S.;Jiang, W.
目的探讨miR-223对NLRP3的影响，进而调节NLRP3/CASP1炎症小体途径介导的促炎细胞因子IL-1 β和IL-18在人牙髓成纤维细胞（HDPFs）中的产生。方法从阻生第三磨牙上获得人牙髓组织（HDPT）和HDPFs。使用miR-223模拟物和抑制剂或NLRP3质粒分别上调或下调hdpf中的miR-223或NLRP3。通过TargetScan 5.1和荧光素酶报告试验进行计算预测以确认目标关联。采用qRT-PCR和Western blotting分别检测NLRP3、caspase-1、IL-1 β和IL-18 mRNA和蛋白的表达。ELISA法检测IL-1 β和IL-18的释放情况。采用单因素方差分析确定实验组与对照组之间差异的显著性；P < 0.05为有统计学意义。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表达降低，NLRP3表达升高（P < 0.05）。计算预测和荧光素酶报告试验证实NLRP3是hdpf中miR-223的直接靶标。与ATP + lps诱导组相比，miR-223抑制剂进一步促进ATP + lps诱导的NLRP3/CASP1炎症小体通路的激活（P < 0.05）。相比之下，与ATP + LPS诱导组相比，miR-223模拟物显著抑制ATP + LPS诱导的NLRP3/CASP1炎性体通路激活（P < 0.05）。结论MiR-223通过靶向NLRP3参与NLRP3/CASP1炎性小体通路介导的促炎细胞因子的产生和分泌，发挥负调控作用。这些数据提供了mirna对NLRP3炎症小体的潜在调节作用的见解，从而为未来的牙髓治疗开辟了新的潜在治疗途径。
Aim To investigate the effect of miR-223 on NLRP3, subsequently regulating the production of the NLRP3/CASP1 inflammasome pathway-mediated proinflammatory cytokines IL-1 beta 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-1 beta and IL-18 was determined by qRT-PCR and Western blotting, respectively. The release of IL-1 beta 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.