MiR-146a-5p Mimic Inhibits NLRP3 Inflammasome Downstream Inflammatory Factors and CLIC4 in Neonatal Necrotizing Enterocolitis.
MiR-146a-5p Mimic Inhibits NLRP3 Inflammasome Downstream Inflammatory Factors and CLIC4 in Neonatal Necrotizing Enterocolitis.
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MiR-146a-5p 模拟物抑制新生儿坏死性小肠结肠炎中的 NLRP3 炎性体下游炎症因子和 CLIC4
DOI：
10.3389/fcell.2020.594143
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发表时间：
2020
影响因子：
5.5
通讯作者：
Lv Z
中科院分区：
文献类型：
--
作者：
Chen J;Chen T;Zhou J;Zhao X;Sheng Q;Lv Z
目的：坏死性小肠结肠炎(NEC)是一种以严重的炎症风暴、肠坏死和穿孔为特征的胃肠道急症。MicroRNA-146a-5p(miR-146a-5p)在多种肠炎性疾病中具有重要的抗炎作用。然而，miR-146a-5p在NEC中的作用及其对核苷酸结合区和富含亮氨酸重复序列的蛋白3(NLRP3)炎症体的影响及其下游炎症因子尚不清楚。本研究旨在探讨miR-146a-5p和NLRP3炎症体及其下游炎症因子在NEC发生发展中的作用。方法：检测肠组织中miR-146a和NLRP3炎症体的表达水平。其次，探讨了miR-146a-5p在体外调节THP-1细胞NLRP3炎性小体激活的机制。最后，为了确定miR-146a-5p对NEC的体内作用，在NEC发生之前，将miR-146a-5p过表达的腺病毒感染NEC小鼠。结果：NLRP3炎性小体酶蛋白caspase-1及其下游炎症因子在人和小鼠NEC肠道标本中均有表达，miR-146a-5p表达水平升高，且主要表达于患肠的巨噬细胞。在体外，只有miR-146a-5p模拟物能抑制THP-1细胞膜上NLRP3炎症性下游炎症因子及其上游蛋白氯细胞内通道蛋白4(CLIC4)的表达，且这种抑制作用仅在轻/中浓度下发生。在体内，MIR-146a-5p过表达的腺病毒可降低CLIC4细胞膜的表达，并抑制NLRP3下游因子的增加。将miR-146a-5p重组腺病毒导入NEC小鼠体内，可提高小鼠的存活率，改善肠道损伤。结论：miR-146a-5p抑制NLRP3炎症体下游炎症因子和CLIC4在NEC膜上的表达。此外，miR-146a-5p可减轻NEC感染肠段的炎症反应和肠道损伤。
Objective: Necrotizing enterocolitis (NEC) is a gastrointestinal emergency with a severe inflammation storm, intestinal necrosis, and perforation. MicroRNA-146a-5p (miR-146a-5p) has been reported to be a valuable anti-inflammatory factor in various intestinal inflammatory disorders. However, the role of miR-146a-5p in NEC, its effects on nucleotide-binding domain and leucine-rich repeat-containing protein 3 (NLRP3) inflammasome, and its downstream inflammatory factors remain unknown. This study aimed to investigate the role of miR-146a-5p and NLRP3 inflammasome and its downstream inflammatory factors in NEC development. Methods: The expression levels of miR-146a and NLRP3 inflammasome were investigated in intestinal tissues. Next, the mechanism by which miR-146a-5p regulates NLRP3 inflammasome activation was explored in vitro in THP-1 cells. Finally, to identify the effects of miR-146a-5p on NEC in vivo, NEC mice were transinfected with miR-146a-5p overexpression adenovirus before the occurrence of NEC. Results: NLRP3 inflammasome enzymatic protein caspase-1 and its downstream inflammatory factors increased in NEC intestinal samples in both humans and mice, and miR-146a-5p expression level was increased and mainly expressed in the macrophages of the affected intestine. In vitro, only miR-146a-5p mimic inhibited NLRP3 inflammasome downstream inflammatory factors and its upstream protein chloride intracellular channel protein 4 (CLIC4) expression in cellular membrane in the THP-1 cell line, and this only occurred under mild/moderate LPS concentration. MiR-146a-5p overexpression adenovirus transfection reduced CLIC4 cellular membrane expression and inhibited NLRP3 downstream factors increasing in vivo. After the transfection of miR-146a-5p adenovirus, the survival rate of NEC mice was increased, and intestinal injury was ameliorated. Conclusion: MiR-146a-5p inhibited NLRP3 inflammasome downstream inflammatory factors and CLIC4 membrane expression in NEC. Additionally, miR-146a-5p could attenuate inflammation and intestinal injury in the NEC-affected intestine.