Jing-Fang Xiang1, Jian-Chun Yu1, Jian-You Zhu2. 1. Department of Emergency, Linzi District People's Hospital, Binzhou Medical University, Zibo, 255400, Shandong, China. 2. Department of Pathology, Linzi District People's Hospital, Binzhou Medical University, Linzi District, Zibo, 255400, Shandong, China. faxiangyihao@163.com.
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.
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.
Authors: Ana Chinchilla; Estefania Lozano; Houria Daimi; Francisco J Esteban; Colin Crist; Amelia E Aranega; Diego Franco Journal: Cardiovasc Res Date: 2010-08-24 Impact factor: 10.787
Authors: Ane M Salvador; Tania Nevers; Francisco Velázquez; Mark Aronovitz; Bonnie Wang; Ana Abadía Molina; Iris Z Jaffe; Richard H Karas; Robert M Blanton; Pilar Alcaide Journal: J Am Heart Assoc Date: 2016-03-15 Impact factor: 5.501