MicroRNA-9 Inhibits NLRP3 Inflammasome Activation in Human Atherosclerosis Inflammation Cell Models through the JAK1/STAT Signaling Pathway.
OBJECTIVE
MicroRNA-9 (miR-9) is involved in inflammatory reaction in atherosclerosis; however, its function and regulatory mechanisms remain unclear. We aimed to uncover the exact roles of miR-9 and downstream signaling pathways using in vitro human atherosclerosis models.


METHODS
We used oxidized low-density lipoprotein (oxLDL)-stimulated human THP-1 derived macrophages, oxLDL-stimulated human primary peripheral blood monocytes and lipopolysaccharides (LPS) or Alum-stimulated human THP-1 derived macrophages as in vitro atherosclerosis inflammation models. Transient transfection of over-expression vectors, small interference RNAs (siRNAs) or antisense oligonucleotides was used to regulate intracellular protein or miR-9 levels. Cell responses and signal transduction were detected by multiple assays including Western blotting, enzyme-linked immunosorbent assay (ELISA) and luciferase reporter assay.


RESULTS
MiR-9 inhibited while anti-miR-9 antisense oligonucleotides induced interleukin-1 beta (IL-1β) and NLRP3 inflammasome activation in all in vitro models. Janus kinase 1 (JAK1) and matrix metalloproteinase 13 (MMP-13) were identified as the target genes of miR-9. In oxLDL-stimulated human THP-1 derived macrophages, knockdown of JAK1 by siRNA blocked the phosphorylation of signal transducer and activator of transcription 1 (STAT1) and mimicked the effects of miR-9. In the same model, JAK1 knockdown blocked the phosphorylation of NF-κB p65 in the nuclei and the phosphorylation of NF-κB IκBα in the cytoplasm.


CONCLUSIONS
Our study demonstrated that miR-9 could inhibit activation of the NLRP3 inflammasome and attenuate atherosclerosis-related inflammation, likely through the JAK1/STAT1 signaling pathway. Therefore, miR-9 may serve as a potential therapeutic target for atherosclerosis.
Department of Cardiology, No. 9 People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Yue
Y
Wang
Department of Cardiology, No. 9 People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Zhihua
Z
Han
Department of Cardiology, No. 9 People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Yuqi
Y
Fan
Department of Cardiology, No. 9 People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Junfeng
J
Zhang
Department of Cardiology, No. 9 People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Kan
K
Chen
Department of Cardiology, No. 9 People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Lin
L
Gao
Department of Cardiology, No. 9 People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Huasu
H
Zeng
Department of Cardiology, No. 9 People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.|Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, China.
Jiatian
J
Cao
Department of Cardiology, No. 9 People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Changqian
C
Wang
D050197:Atherosclerosis / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D006801:Humans; D058847:Inflammasomes / Q000235:genetics / Q000378:metabolism*; D007249:Inflammation / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D053613:Janus Kinase 1 / Q000235:genetics / Q000378:metabolism*; D008077:Lipoproteins, LDL / Q000633:toxicity; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics; D008954:Models, Biological; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein / Q000235:genetics / Q000378:metabolism*; D050794:STAT1 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction
D058847:Inflammasomes; D008077:Lipoproteins, LDL; C541874:MIRN92 microRNA, human; D035683:MicroRNAs; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C453881:NLRP3 protein, human; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; C099252:oxidized low density lipoprotein; C507904:JAK1 protein, human; D053613:Janus Kinase 1
Atherosclerosis; JAK1; NLRP3 inflammasome; STAT1; miR-9