miR-330-5p inhibits NLRP3 inflammasome-mediated myocardial ischaemia–reperfusion injury by targeting TIM3

• Published in: Cardiovascular drugs and therapy 2021-08, Vol.35 (4), p.691-705
• Main Authors: Zuo, Wei, Tian, Ran, Chen, Qian, Wang, Lun, Gu, Qing, Zhao, Hongmei, Huang, Chunmei, Liu, Yingxian, Li, Jingyi, Yang, Xinglin, Xu, Lihong, Zhang, Bo, Liu, Zhenyu
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis; Biomarkers; Biomarkers - metabolism; Cardiology; Cardiomyocytes; Caspase-1; Cerebral blood flow; Coronary artery; Deprivation; Disease Models, Animal; Echocardiography; Echocardiography - methods; Evaluation; Flow cytometry; Heart Failure - diagnosis; Heart Failure - etiology; Heart Failure - metabolism; Hepatitis A Virus Cellular Receptor 2 - metabolism; IL-1β; Immunoblotting; Immunofluorescence; Inflammasomes; Inflammation; Inflammation - metabolism; Injuries; Interleukin 18; Ischemia; Lipopolysaccharides; Lymphocytes; Lymphocytes T; Medicine; Medicine & Public Health; Mice; MicroRNAs - metabolism; Mucin; Myocardial Reperfusion Injury - immunology; Myocardial Reperfusion Injury - metabolism; Myocardial Reperfusion Injury - physiopathology; Occlusion; Original Article; Proteins; Receptors, Cell Surface - metabolism; Regulatory mechanisms (biology); Reperfusion; Signal Transduction; siRNA; Staining; Triphenyltetrazolium chloride; Tumor necrosis factor-α
• ISSN: 0920-3206; 1573-7241
• DOI: 10.1007/s10557-020-07104-8

Background/aims The Nod-like receptor protein-3 (NLRP3) inflammasome signalling pathway is involved in the inflammatory reaction of myocardial ischaemia–reperfusion (I/R) injury. Our previous study showed that miR-330-5p was differentially expressed in both cerebral and myocardial I/R injury, and thus might be a biomarker for I/R injury-related diseases. Another study also indicated that miR-330-5p could promote NLRP3 inflammasome activation in renal IRI. However, the role of miR-330-5p in myocardial I/R injury-induced inflammatory responses is unknown. This study aimed to investigate the role of miR-330-5p in NLRP3 inflammasome-mediated myocardial I/R injury. Methods Myocardial I/R injury was induced in mice by occlusion of the left anterior descending coronary artery for 45 min followed by reperfusion. For NLRP3 inflammasome stimulation in vitro, cardiomyocytes were treated with 2 h of oxygen and glucose deprivation (OGD) or LPS (100 ng/ml). Myocardial miR-330-5p expression was examined by PCR at different treatment times. A miR-330-5p antagomir and an agomir were used to regulate miR-330-5p expression. To evaluate the role of miR-330-5p in myocardial I/R injury, 2,3,5-triphenyltetrazolium chloride (TTC) staining, echocardiography, and immunoblotting were used to assess infarct volume, cardiac function, and NLRP3 inflammasome activation respectively. A luciferase binding assay was used to examine whether miR-330-5p could directly bind to the T cell immunoglobulin domain and mucin domain-containing molecule-3 (TIM3). Finally, the role of the miR-330-5p/TIM3 axis in regulating apoptosis and NLRP3 inflammasome formation was evaluated with flow cytometry assays and immunofluorescence staining. Results Compared to that in the model group, the inhibition of miR-330-5p significantly aggravated myocardial I/R injury, resulting in increased infarct volume and more severe cardiac dysfunction. Moreover, inhibition of miR-330-5p significantly increased the levels of NLRP3 inflammasome-related proteins, including caspase-1, IL-1β, IL-18 and TNF-α, in both in-vivo and in-vitro models. Furthermore, TIM3 was confirmed as a potential target of miR-330-5p. As predicted, suppression of TIM3 by siRNA ameliorated the anti-miR-330-5p-mediated activation of the NLRP3 inflammasome induced by OGD and LPS, thus decreasing cardiomyocyte apoptosis. Conclusions Our study indicated that the miR-330-5p/TIM3 axis was involved in the regulatory mechanism of NLRP3 inflammasome-mediated m