Preliminary in vitro analysis of mechanism of cardiac microvascular endothelial barrier function.
To preliminarily clarify the mechanism of cardiac microvascular endothelial barrier function leading to heart failure, primary HMVEC-D cells were selected and cultured for amplification. The cells were infected with adenovirus vector containing the ADP-ribosylation factor 6 (Arf6) Q67L gene. Full-length and functional fragments of myeloid differentiation primary response 88 and ARF nucleotide-binding site opener genes were established and transfected into HEK293T cells. GTP-Arf6 pull-down experiment, fluorescent quantitative real-time PCR, immuno-coprecipitation, and transendothelial electrical resistance analysis were conducted. Interleukin-1β (IL-1β) induced increase in vascular permeability, whereas inhibitor SC514 blocked IL-1β-induced transfer of nuclear factor-κB into the nucleus, from the cytoplasm. Increase in amount of activated Arf6 promoted reduction in transendothelial electrical resistance. In addition, SecinH3 significantly inhibited increase in vascular permeability, and the progression of heart failure was significantly relieved. Cardiac microvascular endothelial barrier function can lead to heart failure. However, IL-1β induced increase in vascular permeability, which nullified the function of cardiac microvascular endothelial barrier. These findings are closely related to the activation of the Arf6-VE-cadherin signaling pathway.
Intensive Care Unit, Jining No. 1 People's Hospital, Jining, Shandong Province, China.
W
W
Dong
Department of Endocrinology, Jining No. 1 People's Hospital, Jining, Shandong Province, China.
G N
GN
Zhang
Department of Nursing, Jining No. 1 People's Hospital, Jining, Shandong Province, China gaoshuhongdn@163.com.
S H
SH
Gao
D000089982:ADP-Ribosylation Factor 6; D020727:ADP-Ribosylation Factors / Q000378:metabolism; D002199:Capillary Permeability / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D057809:HEK293 Cells; D006333:Heart Failure / Q000378:metabolism / Q000503:physiopathology; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology; D055806:Microvessels / Q000166:cytology*; D009206:Myocardium / Q000166:cytology*
D000089982:ADP-Ribosylation Factor 6; D053583:Interleukin-1beta; D020727:ADP-Ribosylation Factors; C000715348:ARF6 protein, human