title: Hypoxia/ischemia promotes CXCL10 expression in cardiac microvascular endothelial cells by NFkB activation.
abstract: CXCL10, the chemokine with potent chemotactic activity on immune cells and other non-immune cells expressing its receptor CXCR3, has been demonstrated to involve in myocardial infarction, which was resulted from hypoxia/ischemia. The cardiac microvascular endothelial cells (CMECs) are the first cell type which is implicated by hypoxia/ischemia. However, the potential molecular mechanism by which hypoxia/ischemia regulates the expression of CXCL10 in CMECs remains unclear. In the present study, the expression of CXCL10 was firstly examined by real-time PCR and ELISA analysis. Several potential binding sites (BS) for transcription factors including NF-kappaB (NFkB), HIF1 alpha (HIF1α) and FoxO3a were identified in the promoter region of CXCL10 gene from -2000 bp to -1 bp using bioinformatics software. Luciferase reporter gene vectors for CXCL10 promoter and for activation of above transcription factors were constructed. The activation of NFkB, hypoxia-inducible transcription factor-1 alpha (HIF-1α) and FoxO3a was also analyzed by Western blotting. It was shown that the production of CXCL10 in CMECs was significantly increased by hypoxia/ischemia treatment, in parallel with the activation of CXCL10 promoter examined by reporter gene vector system. Furthermore, transcription factors including NFkB, HIF1α and FoxO3a were activated by hypoxia/ischemia in CMECs. However, over-expression of NFkB, but not that of HIF1α or FoxO3a, significantly promoted the activation of CXCL10 promoter reporter gene. These findings indicated that CXCL10 production in CMECs was significantly increased by hypoxia/ischemia, at least in part, through activation of NFkB pathway and subsequently binding to CXCL10 promoter, finally promoted the transcription of CXCL10 gene.
authors.affiliation: Key Laboratory of Regenerative Medicine (JNU-CUHK), Ministry of Education and Department of Developmental & Regenerative Biology, Ji Nan University, Guangzhou 510632, China.
authors.forename: Jing-Bo
authors.initials: JB
authors.lastname: Xia
authors.affiliation: Key Laboratory of Regenerative Medicine (JNU-CUHK), Ministry of Education and Department of Developmental & Regenerative Biology, Ji Nan University, Guangzhou 510632, China.
authors.forename: Guang-Hui
authors.initials: GH
authors.lastname: Liu
authors.affiliation: Key Laboratory of Regenerative Medicine (JNU-CUHK), Ministry of Education and Department of Developmental & Regenerative Biology, Ji Nan University, Guangzhou 510632, China.
authors.forename: Zhuo-Ying
authors.initials: ZY
authors.lastname: Chen
authors.affiliation: Key Laboratory of Regenerative Medicine (JNU-CUHK), Ministry of Education and Department of Developmental & Regenerative Biology, Ji Nan University, Guangzhou 510632, China.
authors.forename: Cheng-Zhou
authors.initials: CZ
authors.lastname: Mao
authors.affiliation: Key Laboratory of Regenerative Medicine (JNU-CUHK), Ministry of Education and Department of Developmental & Regenerative Biology, Ji Nan University, Guangzhou 510632, China.
authors.forename: Deng-Cheng
authors.initials: DC
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Regenerative Medicine (JNU-CUHK), Ministry of Education and Department of Developmental & Regenerative Biology, Ji Nan University, Guangzhou 510632, China.
authors.forename: Hai-Yan
authors.initials: HY
authors.lastname: Wu
authors.affiliation: Department of Physiology, Yonsei University Wonju College of Medicine, Wonju, Gangwon 220-701, Republic of Korea.
authors.forename: Kyu-Sang
authors.initials: KS
authors.lastname: Park
authors.affiliation: School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong Special Administrative Region.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Department of Microbiology, Yonsei University Wonju College of Medicine, Wonju, Gangwon 220-701, Republic of Korea.
authors.forename: Soo-Ki
authors.initials: SK
authors.lastname: Kim
authors.affiliation: Key Laboratory of Regenerative Medicine (JNU-CUHK), Ministry of Education and Department of Developmental & Regenerative Biology, Ji Nan University, Guangzhou 510632, China. Electronic address: tdongbme@jnu.edu.cn.
authors.forename: Dong-Qing
authors.initials: DQ
authors.lastname: Cai
authors.affiliation: Key Laboratory of Regenerative Medicine (JNU-CUHK), Ministry of Education and Department of Developmental & Regenerative Biology, Ji Nan University, Guangzhou 510632, China. Electronic address: qixufeng@jnu.edu.cn.
authors.forename: Xu-Feng
authors.initials: XF
authors.lastname: Qi
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D015153:Blotting, Western; D015687:Cell Hypoxia; D002478:Cells, Cultured; D054357:Chemokine CXCL10 / Q000235:genetics / Q000378:metabolism*; D003331:Coronary Vessels / Q000166:cytology*; D042783:Endothelial Cells / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D000071316:Forkhead Box Protein O3 / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism; D007511:Ischemia; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D054357:Chemokine CXCL10; C499515:FOXO3 protein, rat; D000071316:Forkhead Box Protein O3; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D016328:NF-kappa B
keywords: CMECs; CXCL10; Expression; Hypoxia/ischemia; NFkB