ABSTRACT
BACKGROUND: MiR-27 has been found to present an overt myocardial expression during cardiogenesis. However, whether miR-27 involves in myocarditis development and the possible molecular mechanism remain unknown. The purpose of this study was to investigate the biological characteristic of miR-27 in LPS-damaged H9c2 cells. METHODS: H9c2 cells were treated with lipopolysaccharide (LPS, 10 µg/ml) for 12 h to form cell injury. MiR-27 mimic and inhibitor were used to up-regulate or down-regulate miR-27 expression. MTT assay and flow cytometry analysis were conducted to test cell viability and apoptosis. The relative RNA expression level of miR-27 and intercellular adhesion molecule 1 (ICAM1) was determined by qRT-PCR. Luciferase reporter gene assay was utilized to confirm the interaction between miR-27 and ICAM1. Western blot was used to determine the protein expression levels. RESULTS: We observed that LPS treatment significantly decreased the level of miR-27 in H9c2 cells. Moreover, LPS exposure suppressed cell viability, promoted cell apoptosis and increased the relative expression of p-NF-κB p65/NF-κB p65 and p-IκBα/IκBα. Up-regulation of miR-27 increased cell proliferation and reduced cell apoptosis, while down-regulation of miR-27 suppressed cell growth and promoted cell apoptosis. ICAM1 was predicted and verified as a target of miR-27, and the expression of ICAM1 is negatively regulated by miR-27. The relative expression of p-NF-κB p65/NF-κB p65 and p-IκBα/IκBα was dramatically decreased by miR-27 mimic and increased by miR-27 inhibitor. CONCLUSION: Our study illustrated that up-regulation of miR-27 exhibits a protective effect on LPS-damaged H9c2 cells, which may be achieved by regulating ICAM1 and NF-κB signaling.
Subject(s)
Intercellular Adhesion Molecule-1/genetics , Lipopolysaccharides/toxicity , MicroRNAs/metabolism , Myocytes, Cardiac/metabolism , Animals , Cell Line , Gene Expression Regulation , Intercellular Adhesion Molecule-1/metabolism , Myocytes, Cardiac/drug effects , NF-kappa B/antagonists & inhibitors , Rats , Up-RegulationABSTRACT
The aim of the study is to explore the effects and mechanism of the action of ligustrazine on isoprenaline-induced cardiomyocyte hypertrophy. Primary culture of neonatal rat cardiomyocytes was used as the model, and isoprenaline was used to induce cardiomyocyte hypertrophy. Effects of different dosages of ligustrazine polysaccharide on the cardiomyocyte were observed. RT-PCR was used to detect the expression of atrial natriuretic factor (ANP) mRNA, and Western blot analysis was used to detect the CaN protein level in cardiomyocytes. After treating with ligustrazine, the significant increase of MDA content and decrease of SOD activity were inhibited in supernatant. Compared to the control group, ANP mRNA in isoprenaline-treated cardiomyocytes was significantly increased (P < 0.05); compared to the isoprenaline group, ANP mRNA was significantly decreased in all ligustrazine groups (P < 0.01). In all ligustrazine groups, the CaN expression was inhibited in isoprenaline-treated cardiomyocytes in a dose-dependent manner. In conclusion, ligustrazine has protective effects on isoprenaline-induced neonatal rat cardiomyocyte, which may be related to the decrease of CaN expression.
