Nobels days 2013
1
Contact Name : George Gkoumas Email : [email protected] Gender difference in inflammatory disease such as CVD has been reported both in human and experimental animals. The gender specific role of estrogen has been reflected in vascular injury response. Since observational studies have shown substantial benefits of estrogen in reducing the relative risk of Coronary heart disease (CHD) by 50%, we examined the effects of estradiol on NLRP3 inflammasome components and inflammatory cytokines in vascular cells. Introduction The present study indicates that 17β-estradiol may play an important role in the regulation of vascular inflammation both by modulating the NLRP3 inflammasome components and proinflammatory cytokines such as IL-6 and IL-8. Down regulation of NLRP3 inflammasome and proinflammatory cytokines might be one of the mechanisms of the protective effect of estrogen on cardiovascular system. Results References 17β-estradiol reduces vascular inflammation through the regulation of NLRP3 inflammasome in HUVEC and AOSMC George Gkoumas 1, 2 , Geena Paramel Varghese 2 , Karin fransén 2 , Allan sirsjö 2 1. Varghese G, Fransén K, Hurtig‑Wennlöf A, Bengtsson T, Jansson J, Sirsjö A. (2013) .”Q705K variant in NLRP3 gene confers protection against myocardial infarction in female individuals." Biomedical Reports 1(6): 879-882. 2. Xing D, Susan N, Yiu-Fai C, Fadi H. (2009) . "Estrogen and mechanisms of vascular protection." Arterioscler Thromb Vasc Biol 29(3): 289-295. 1 Department of Medical Laboratories, Biomedical Laboratory Technologist – Technological Educational Institute of Athens, Greece. 2 School of Health and Medical Sciences, Örebro University, Sweden 10/12/2013. AOSMC and HUVEC were grown in estrogen free medium before the assay. Due to the low basal expression of NLRP3 inflammasome components in HUVEC, the effect of estrogen was examined by stimulating the cells with TNFα. Real time PCR and ELISA was performed to analyze the mRNA expression and IL-1β, IL-6 and IL-8 protein. Methods Exogenous 17β -estradiol significantly down regulated the expression of NLRP3, caspase-1 and IL-1β in HUVEC and AOSMC. In AOSMC, the treatment of estrogen resulted in a moderate reduction of IL-1β release and significant reduction of IL-6 and IL-8 levels. cntrl(OH) tnf(OH) tnf+estrogen estrogen 0 10 20 30 40 50 60 70 80 Relative expression of NLRP3 in HUVEC Relative value cntrol tnf tnf+estrogen estrogen 0 0.2 0.4 0.6 0.8 1 1.2 1.4 Relative expression of caspase-1 in HUVEC Relative value cntrl(OH) tnf(OH) tnf+estrogen estrogen 0 1 2 3 4 5 6 7 8 Relative expression of IL-1β in HUVEC Relative value control Tnf Tnf+Estrogen Estrogen 0 2000 4000 6000 8000 10000 12000 14000 16000 IL-8 levels in HUVEC pg/ml Fig 4: IL-8 levels in the HUVEC culture medium control tnf tnf+Estrogen Estrogen 0 200 400 600 800 1000 1200 1400 IL-6 levels in AOSMC pg/ml Fig 5: IL-6 levels in the AOSMC culture medium control tnf tnf+Estrogen Estrogen 0 10000 20000 30000 40000 50000 60000 70000 IL-8 levels in AOSMC pg/ml Fig 6: IL-8 levels in the AOSMC culture medium cntrl tnf tnf+estrogen estrogen 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 IL-1β relative expression Relative value cntrl tnf tnf+estrogen estrogen 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 Caspase-1 relative expression Relative value Fig 7: Relative expression of Caspase-1 in Aortic Smooth Muscle Cells after normalization with a house-keeping gene (GAPDH) * ** * ** * * * ** Conclusion Figure 2: Plaque progression. Figure 1: Pathway for the activation of NLRP3 inflammasome complex: The activation can be triggered by many factors such as crystalline stucture, muramyldipeptide (MDP), ATP, reactive oxygen species (ROS) and pathogen associated molecular patterns (PAMPS). The NLRP3 inflammasome activates IL-1β via pro-caspase 1, triggering the activation of caspase 1 and the maturation and secretion of pro-inflammatory cytokines such as interleukin-1β (IL-1β) and IL-18. Figure 3: Plaque rupture and thrombosis. Fig 2: Relative expression of Caspase 1 in Human Umbilical Vein Endothelial Cells after normalization with a house-keeping gene (Cyclophilin A) Fig 1: Relative expression of NLRP3 in Human Umbilical Vein Endothelial Cells after normalization with a house-keeping gene (Cyclophilin A) Fig 3: Relative expression of IL-1β in Human Umbilical Vein Endothelial Cells after normalization with a house-keeping gene (Cyclophilin A) Fig 8: Relative expression of IL-1β in Aortic Smooth Muscle Cells after normalization with a house-keeping gene (GAPDH) Aim The aim of this study was to examine the effects of estradiol on NLRP3 inflammasome components and proinflammatory cytokines in vascular cells.
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Transcript of Nobels days 2013
ContactName : George Gkoumas Email : [email protected]
Gender difference in inflammatory disease such as CVD has been reported both in human and experimental animals. The gender specific role of estrogen has been reflected in vascular injury response. Since observational studies have shown substantial benefits of estrogen in reducing the relative risk of Coronary heart disease (CHD) by 50%, we examined the effects of estradiol on NLRP3 inflammasome components and inflammatory cytokines in vascular cells.
Introduction
The present study indicates that 17β-estradiol may play an important role in the regulation of vascular inflammation both by modulating the NLRP3 inflammasome components and proinflammatory cytokines such as IL-6 and IL-8. Down regulation of NLRP3 inflammasome and proinflammatory cytokines might be one of the mechanisms of the protective effect of estrogen on cardiovascular system.
Results
References
17β-estradiol reduces vascular inflammation through the regulation of NLRP3 inflammasome in HUVEC and AOSMC
George Gkoumas1, 2, Geena Paramel Varghese2, Karin fransén2, Allan sirsjö2
1. Varghese G, Fransén K, Hurtig‑Wennlöf A, Bengtsson T, Jansson J, Sirsjö A. (2013).”Q705K variant in NLRP3 gene confers protection against myocardial infarction in female individuals." Biomedical Reports 1(6): 879-882.
2. Xing D, Susan N, Yiu-Fai C, Fadi H. (2009). "Estrogen and mechanisms of vascular protection." Arterioscler Thromb Vasc Biol 29(3): 289-295.
1Department of Medical Laboratories, Biomedical Laboratory Technologist – Technological Educational Institute of Athens, Greece. 2School of Health and Medical Sciences, Örebro University, Sweden 10/12/2013.
AOSMC and HUVEC were grown in estrogen free medium before the assay. Due to the low basal expression of NLRP3 inflammasome components in HUVEC, the effect of estrogen was examined by stimulating the cells with TNFα. Real time PCR and ELISA was performed to analyze the mRNA expression and IL-1β, IL-6 and IL-8 protein.
Methods
Exogenous 17β -estradiol significantly down regulated the expression of NLRP3, caspase-1 and IL-1β in HUVEC and AOSMC. In AOSMC, the treatment of estrogen resulted in a moderate reduction of IL-1β release and significant reduction of IL-6 and IL-8 levels.
cntrl(OH) tnf(OH) tnf+estrogen estrogen0
10
20
30
40
50
60
70
80Relative expression of NLRP3 in HUVEC
Rel
ativ
e va
lue
cntrol tnf tnf+estrogen estrogen0
0.2
0.4
0.6
0.8
1
1.2
1.4Relative expression of caspase-1 in HUVEC
Rel
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lue
cntrl(OH) tnf(OH) tnf+estrogen estrogen0
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2
3
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Relative expression of IL-1β in HUVEC
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lue
control Tnf Tnf+Estrogen Estrogen0
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IL-8 levels in HUVEC
pg/m
lFig 4: IL-8 levels in the HUVEC culture medium
control tnf tnf+Estrogen Estrogen0
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IL-6 levels in AOSMC
pg/m
l
Fig 5: IL-6 levels in the AOSMC culture medium
control tnf tnf+Estrogen Estrogen0
10000
20000
30000
40000
50000
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IL-8 levels in AOSMC
pg/m
l
Fig 6: IL-8 levels in the AOSMC culture medium
cntrl tnf tnf+estrogen estrogen0
0.2
0.4
0.6
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1.6IL-1β relative expression
Rel
ativ
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cntrl tnf tnf+estrogen estrogen0
0.2
0.4
0.6
0.8
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1.2
1.4
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Caspase-1 relative expression
Rel
ativ
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lue
Fig 7: Relative expression of Caspase-1 in Aortic Smooth Muscle Cells after normalization with a house-keeping gene (GAPDH)
*
***
**
*
*
*
**
Conclusion
Figure 2: Plaque progression.
Figure 1: Pathway for the activation of NLRP3 inflammasome complex: The activation can be triggered by many factors such as crystalline stucture, muramyldipeptide (MDP), ATP, reactive oxygen species (ROS) and pathogen associated molecular patterns (PAMPS). The NLRP3 inflammasome activates IL-1β via pro-caspase 1, triggering the activation of caspase 1 and the maturation and secretion of pro-inflammatory cytokines such as interleukin-1β (IL-1β) and IL-18.
Figure 3: Plaque rupture and thrombosis.
Fig 2: Relative expression of Caspase 1 in Human Umbilical Vein Endothelial Cells after normalization with a house-keeping gene (Cyclophilin A)
Fig 1: Relative expression of NLRP3 in Human Umbilical Vein Endothelial Cells after normalization with a house-keeping gene (Cyclophilin A)
Fig 3: Relative expression of IL-1β in Human Umbilical Vein Endothelial Cells after normalization with a house-keeping gene (Cyclophilin A)
Fig 8: Relative expression of IL-1β in Aortic Smooth Muscle Cells after normalization with a house-keeping gene (GAPDH)
Aim
The aim of this study was to examine the effects of estradiol on NLRP3 inflammasome components and proinflammatory cytokines in vascular cells.
