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MicroRNA-325-3p protects the heart after myocardial infarction by inhibiting RIPK3 and programmed necrosis in mice.
BMC Molecular Biology ( IF 4.619 ) Pub Date : 2019-06-27 , DOI: 10.1186/s12867-019-0133-z
Dong-Ying Zhang 1 , Bing-Jian Wang 1 , Min Ma 2 , Kun Yu 1 , Qing Zhang 1 , Xi-Wen Zhang 1
Affiliation  

Receptor-interacting serine-threonine kinase 3 (RIPK3)-mediated necroptosis has been implicated in the progression of myocardial infarction (MI), but the underlying mechanisms, particularly whether microRNAs (miRNAs) are involved, remain largely unknown. A microarray analysis was used to screen for miR-325-3p expression in myocardial tissues from MI mice, and the expression was confirmed with qRT-PCR. The levels of myocardial enzymes were measured using commercial kits, and an echocardiography system was utilized for the detection of cardiac function parameters. The pathological features and infarction sizes of cardiac tissues were examined using H&E, TCC and Masson’s trichrome staining, and the amount of cell apoptosis was determined using an in situ TUNEL assay. Cardiomyocytes were isolated and then subjected to hypoxia induction in vitro. The expression of the RIPK1, RIPK3 and phosphorylated MLKL (p-MLKL) proteins was measured using a Western blot. The mouse cardiomyocyte cell viability was analyzed by an MTT assay. The mRNA target of miR-325-3p was predicted using TargetScan v7.2 and then validated using a dual-luciferase reporter assay. The overexpression of miR-325-3p evidently decreased the expression levels of lactate dehydrogenase (LDH), phosphocreatine kinase (CK), superoxide dismutase (SOD) and malondialdehyde (MDA), inhibited left ventricular end-diastolic diameter (LVEDD) and left ventricular end-systolic diameter (LVESD), and promoted left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVES). In addition, miR-325-3p overexpression attenuated the degree of injury to the cardiac tissue, decreased the infarct sizes and downregulated the expression of the necrosis-related proteins RIPK1, RIPK3 and p-MLKL. The RIPK1/RIPK3/p-MLKL axis-induced necroptosis that occurred during MI was mediated by a miRNA module, miR-325-3p, which can effectively ameliorate the symptoms of MI by suppressing the expression of RIPK3.

中文翻译:

MicroRNA-325-3p通过抑制小鼠的RIPK3和程序性坏死来保护心肌梗塞后的心脏。

受体相互作用的丝氨酸-苏氨酸激酶3(RIPK3)介导的坏死病与心肌梗死(MI)的进展有关,但其潜在机制,特别是是否涉及microRNA(miRNA),仍然是未知的。使用微阵列分析筛选来自MI小鼠的心肌组织中miR-325-3p的表达,并通过qRT-PCR确认表达。使用商业试剂盒测量心肌酶的水平,并使用超声心动图系统检测心脏功能参数。使用H&E,TCC和Masson三色染色检查心脏组织的病理特征和梗塞大小,并使用原位TUNEL测定法测定细胞凋亡的数量。分离心肌细胞,然后在体外进行缺氧诱导。使用蛋白质印迹法测量RIPK1,RIPK3和磷酸化的MLKL(p-MLKL)蛋白的表达。通过MTT测定法分析小鼠心肌细胞的生存力。使用TargetScan v7.2预测miR-325-3p的mRNA靶标,然后使用双荧光素酶报告基因检测法对其进行验证。miR-325-3p的过表达明显降低了乳酸脱氢酶(LDH),磷酸肌酸激酶(CK),超氧化物歧化酶(SOD)和丙二醛(MDA)的表达水平,抑制了左心室舒张末期直径(LVEDD)和左心室收缩末期直径(LVESD),并促进左心室射血分数(LVEF)和左心室分数缩短(LVES)。此外,miR-325-3p的过度表达可减轻对心脏组织的损伤程度,降低了梗塞面积并下调了坏死相关蛋白RIPK1,RIPK3和p-MLKL的表达。miRNA发生的miR-325-3p介导了MIK期间发生的RIPK1 / RIPK3 / p-MLKL轴诱导的坏死病,它可以通过抑制RIPK3的表达有效缓解MI的症状。
更新日期:2019-06-27
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