Background/Aims: Ginsenoside Rb1 (GS-Rb1) is one of the most important active pharmacological extracts of the Traditional Chinese Medicine ginseng, with extensive evidence of its cardioprotective properties. Mir-208 has been shown to act as a biomarker of acute myocardial infarction in vivo studies including man. However the impact of miR-208 on the protective effect of GS-Rb1 in hypoxia/ischemia injured cardiomyocytes remains unclear. The current study aims to investigate the target gene of miR-208 and the impact on the protective effect of GS-Rb1 in hypoxia/ischemia (H/I) injuried cardiomyocytes. Materials and Methods: Primary cultures of neonatal rat cardiomyocytes (NRCMs) was subjected to the H/I conditions with or without GS-Rb1. Cell viability was calculated by MTT assay and confirmed by flow cytometry analysis. Mir-208 was then detected by qRT-PCR. Luciferase reporter assay was carried out to detect the target gene of Mir-208. Then the NRCMs were transfected with miR-208 mimics and inhibitors to evaluate the impact on cardioprotective properties of Rb1. Results: The miR-208 expression level was clearly upregulated in the H/I treated NRCMs accompanied by the percentage of the apoptotic cells which could be reversed by GS-Rb1 pretreatment. The nemo-like kinase (NLK) mRNA and protein expression levels were decreased in H/I group measured by RT-PCR and western blotting. Luciferase activity assay was then carried out to identify that NLK may be a direct target of mir-208. MTT assay showed that miR-208 inhibitor slightly decreased the protective effect of Rb1 on the H/I impaired NRCMs. However, results showed no statistical difference. Conclusions: These findings proved that NLK was a direct target of mir-208 and miR-208 act indirectly during Rb1 protecting H/I impaired NRCMs and further researches were needed to explore the relationship that microRNAs and other signal pathways in the protective effect of GS-Rb1 on the hypoxia/ischemia injuries in cardiomyocytes. (C) 2016 The Author(s) Published by S. Karger AG, Basel.
基金：
National Natural Science Foundation of ChinaNational Natural Science Foundation of China [81273890, 81173367]; National key basic research development program (973 Program)National Basic Research Program of China [2015CB554400]; Postdoctoral Science Foundation of Beijing [2013ZZ-57]
第一作者机构：[1]Peking Univ, Hosp 3, Haidian Sect, Beijing Haidian Hosp, 29 Zhongguancun Dajie, Beijing, Peoples R China;[2]Capital Med Univ, Beijing Tiantan Hosp, Beijing Neurosurg Inst, Dept Pathophysiol, Beijing, Peoples R China;
通讯作者：
通讯机构：[1]Peking Univ, Hosp 3, Haidian Sect, Beijing Haidian Hosp, 29 Zhongguancun Dajie, Beijing, Peoples R China;[5]Beijing Haidian Hosp, 29 Zhongguancun Dajie, Beijing 100080, Peoples R China
推荐引用方式(GB/T 7714)：
Yan Xu,Liu Jianxun,Wu Hongjin,et al.Impact of miR-208 and its Target Gene Nemo-Like Kinase on the Protective Effect of Ginsenoside Rb1 in Hypoxia/Ischemia Injuried Cardiomyocytes[J].CELLULAR PHYSIOLOGY AND BIOCHEMISTRY.2016,39(3):1187-1195.doi:10.1159/000447825.
APA：
Yan, Xu,Liu, Jianxun,Wu, Hongjin,Liu, Yuna,Zheng, Sidao...&Yang, Cui.(2016).Impact of miR-208 and its Target Gene Nemo-Like Kinase on the Protective Effect of Ginsenoside Rb1 in Hypoxia/Ischemia Injuried Cardiomyocytes.CELLULAR PHYSIOLOGY AND BIOCHEMISTRY,39,(3)
MLA：
Yan, Xu,et al."Impact of miR-208 and its Target Gene Nemo-Like Kinase on the Protective Effect of Ginsenoside Rb1 in Hypoxia/Ischemia Injuried Cardiomyocytes".CELLULAR PHYSIOLOGY AND BIOCHEMISTRY 39..3(2016):1187-1195
