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Abstracts 1
MICROCIRCULATORY BLOOD FLOW IN HINDLIMBS DURING REMOTE ISCHAEMIC PRECONDITIONING
S Abdul-Ghani, G D Angelini, M Saadeh Suleiman; The Bristol Heart Institute, Faculty of Medicine and Dentistry, University of Bristol, UK 10.1136/heartjnl-2013-305297.1
Background Brief ischaemia of one organ protects the heart against a sustained ischaemia-reperfusion insult (Remote ischaemic preconditioning, RIPC). The nature of the signal released from the remote organ and the timing of the triggered changes in the target organs remain controversial. It is not known whether RIPC also alters blood flow in other organs. In this study we investigated the changes in microcirculatory blood flow (MBF) in both hind limbs during RIPC and measured myocardial metabolites (e.g. ATP, AMP and adenosine) after induction of RIPC. Methods RIPC was induced in C57/Bl6 mice using four cycles of 5 min ischaemia and 5 min reperfusion of the hind limb (experimental limb) using blood pressure cuff. MBF in both RIPC mouse and in control mouse was measured using Laser Doppler Flowmetry throughout the procedure of 50 min. In separate experiment, hearts were excised from RIPC and control mice and extracted for measuring metabolites using HPLC. Results In control mice (no RIPC application) there was no significant change in MBF in both hind limbs. Inflating blood pressure cuff for 5 min resulted in complete occlusion of MBF in the experimental hind limb. Reperfusion in each cycle of RIPC was associated with a hyperaemic response. In contrast, MBF in the un-cuffed hind limb was reduced during the RIPC cycles. Interestingly, HPLC analysis showed that RIPC causes ischaemic stress (increase AMP/ATP). Conclusion Our mouse model of RIPC is associated with reduced MBF in the non-conditioned hind limb and metabolic stress in the myocardium. Supported by NIHR BRISTOL BIOMEDICAL RESEARCH UNIT IN CARDIOVASCULAR MEDICINE.
2
IL-1B SECRETION IN CORONARY VASCULAR ENDOTHELIUM IS MEDIATED BY NEUTROPHIL SERINE PROTEASE AND IS INDEPENDENT OF CASPASE-1
M Alfaidi, H L Wilson, A Burnett, B H Abdul-Aema, V Ridger, J Chamberlain, S E Francis; Department of Cardiovascular Science, University of Sheffield, UK 10.1136/heartjnl-2013-305297.2
Endothelial cells (ECs) are critically involved in the pathogenesis of atherosclerosis by producing inflammatory mediators, including interleukin-1 beta (IL-1b). However, its mechanism of externalisation is yet to be elucidated. This study investigates the effect of neutrophil elastase (NE) on ECs in terms of IL-1b release and explores the underlying mechanism. Human coronary artery endothelial cells (HCAEC) were treated with a combination of tumour necrosis factor-alpha and interleukin-1 alpha then incubated with NE for 2 or 6 h. ELISA, western blotting, flow cytometry and live cell imaging were used. Paraffinembedded sections of aortic sinus lesions of apoe-/- mice were immunostained for NE and VWF (Von Willebrand factor). NE is predominantly expressed in ECs in experimental atherosclerosis. In vitro, NE caused significant IL-1b release into supernatants after 6h (579 ± 90 vs. 80 ± 19 pg/ml in control; n = 4, p < 0.0001). The release was via microparticle shedding which is significantly attenuated by neutrophil elastase inhibitor III (62.64 ± 10 vs. 579.3 ± 90 pg/ml in NE-primed cells; n = 3, p
Abstracts 1
MICROCIRCULATORY BLOOD FLOW IN HINDLIMBS DURING REMOTE ISCHAEMIC PRECONDITIONING
S Abdul-Ghani, G D Angelini, M Saadeh Suleiman; The Bristol Heart Institute, Faculty of Medicine and Dentistry, University of Bristol, UK 10.1136/heartjnl-2013-305297.1
Background Brief ischaemia of one organ protects the heart against a sustained ischaemia-reperfusion insult (Remote ischaemic preconditioning, RIPC). The nature of the signal released from the remote organ and the timing of the triggered changes in the target organs remain controversial. It is not known whether RIPC also alters blood flow in other organs. In this study we investigated the changes in microcirculatory blood flow (MBF) in both hind limbs during RIPC and measured myocardial metabolites (e.g. ATP, AMP and adenosine) after induction of RIPC. Methods RIPC was induced in C57/Bl6 mice using four cycles of 5 min ischaemia and 5 min reperfusion of the hind limb (experimental limb) using blood pressure cuff. MBF in both RIPC mouse and in control mouse was measured using Laser Doppler Flowmetry throughout the procedure of 50 min. In separate experiment, hearts were excised from RIPC and control mice and extracted for measuring metabolites using HPLC. Results In control mice (no RIPC application) there was no significant change in MBF in both hind limbs. Inflating blood pressure cuff for 5 min resulted in complete occlusion of MBF in the experimental hind limb. Reperfusion in each cycle of RIPC was associated with a hyperaemic response. In contrast, MBF in the un-cuffed hind limb was reduced during the RIPC cycles. Interestingly, HPLC analysis showed that RIPC causes ischaemic stress (increase AMP/ATP). Conclusion Our mouse model of RIPC is associated with reduced MBF in the non-conditioned hind limb and metabolic stress in the myocardium. Supported by NIHR BRISTOL BIOMEDICAL RESEARCH UNIT IN CARDIOVASCULAR MEDICINE.
2
IL-1B SECRETION IN CORONARY VASCULAR ENDOTHELIUM IS MEDIATED BY NEUTROPHIL SERINE PROTEASE AND IS INDEPENDENT OF CASPASE-1
M Alfaidi, H L Wilson, A Burnett, B H Abdul-Aema, V Ridger, J Chamberlain, S E Francis; Department of Cardiovascular Science, University of Sheffield, UK 10.1136/heartjnl-2013-305297.2
Endothelial cells (ECs) are critically involved in the pathogenesis of atherosclerosis by producing inflammatory mediators, including interleukin-1 beta (IL-1b). However, its mechanism of externalisation is yet to be elucidated. This study investigates the effect of neutrophil elastase (NE) on ECs in terms of IL-1b release and explores the underlying mechanism. Human coronary artery endothelial cells (HCAEC) were treated with a combination of tumour necrosis factor-alpha and interleukin-1 alpha then incubated with NE for 2 or 6 h. ELISA, western blotting, flow cytometry and live cell imaging were used. Paraffinembedded sections of aortic sinus lesions of apoe-/- mice were immunostained for NE and VWF (Von Willebrand factor). NE is predominantly expressed in ECs in experimental atherosclerosis. In vitro, NE caused significant IL-1b release into supernatants after 6h (579 ± 90 vs. 80 ± 19 pg/ml in control; n = 4, p < 0.0001). The release was via microparticle shedding which is significantly attenuated by neutrophil elastase inhibitor III (62.64 ± 10 vs. 579.3 ± 90 pg/ml in NE-primed cells; n = 3, p
