periodontal pathogen egat21-12-2011ฉบับแก้ไข · cholesterol early study •...
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Periodontal pathogen and cholesterol crystal induce NLRP3 inflammasome-mediated
interleukin-1β secretion in human macrophages
Department of Periodontologyp gyFaculty of Dentistry
Chulalongkorn UniversityChulalongkorn University
Oral cavity as a part of the bodyOral cavity as a part of the body• Human mouth has long been Human mouth has long been
recognized as a infection and
connected to the systemic health
(Miller, 1890)
• European workshop 2010 : from epidemiologic study, there is a
moderate significant association
between periodontal disease and between periodontal disease and
cardiovascular disease (Bouchard
et al., 2010)
2
Periodontal disease-Atherosclerosis relationshipPeriodontal disease Atherosclerosis relationship
Distant chronic infection “periodontitis” is one of p
the candidates
Ulcerated inflamed periodontal pockets
plaque micro-organisms and their products i.e.
LPS
blood vessels in the connective tissue
systemic circulation
activate inflammatory & response of artery(Gibson III et al., 2006)
3
Periodontal disease-Atherosclerosis relationship Periodontal disease Atherosclerosis relationship
Supporting study :pp g y
• key periodontal pathogens such as P. gingivalis and Aggregatibacteractinomycetemcomitans could be detected in human be de ec ed u aatherosclerotic lesion
(Haraszthy et al., 2000)
• P. gingivalis promoted foam cell formation in the presence
f lo densit lipoprotein of low density lipoprotein (Giacona et al., 2004)
4
CholesterolCholesterol
Early study• Cholesterol : hallmark of late
Breakthrough knowledge• Recent study (Duewell et al., 2010
stage of atherosclerotic lesions.
• It could only be observed in the
Nature ) demonstrated that
cholesterol crystals can be It could only be observed in the
mature atherosclerotic lesions or
in the late stage of atherogenesis
detected at an early stage of the
disease development in the
atherosclerosisin the late stage of atherogenesis
(Small, 1988)atherosclerosis
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From Duewell StudyFrom Duewell Study…
• A novel link between cholesterol crystals and inflammation in
atherosclerosis lesion was shown by their ability to activate
caspase-1- activating “nucleotide-binding domain leucine-rich
repeated containing family, pyrin domain containing 3” (NLRP3) repeated containing family, pyrin domain containing 3 (NLRP3) in form of INFLAMMASOMES
• results in cleavage and secretion of Interleukin-1 (IL-1) family
cytokines, in human and mouse macrophages (Rajamaki et
al.,2010; Duewell et al.,2010).
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NOD-like receptorsNOD-like receptorsNLRP3
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NLRP3 inflammasome activationNLRP3 inflammasome activation
(Horvath et al., 2011)
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Interleukin 1βInterleukin-1β
• Important proinflammatory cytokines • Triggers inflammationTriggers inflammation• Macrophage : major source
(Taylor, 2010 )
• Level of IL-1β correlate with disease severityse e ty
( Libby, 2002 )
• Requires 2 separate signals for production( Martinon et al., 2009)
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M1 VS M2 macrophageM1 VS M2 macrophage
Characteristics M1 macrophage M2 macrophage
Synonym Classically activated Alternatively activated
macrophage macrophage
Th polarization Th1 Th2
Stimulating factor GM-CSF M-CSF
Morphology Round shape Elongated shape
Cell surface markers CD14+, CD16-,
CD36-,CD163-
CD14+, CD16+,
CD36+,CD163+
(Boyle, 2005; Waldo et al., 2008; Verreck et al., 2006)
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Macrophage phenotypesMacrophage phenotypes
M1
(Duewell et al 2010)
M1
(Duewell et al., 2010)
M2
(Brocheriou et al., 2011)(Rajamaki et al., 2010)
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From our researchFrom our research…M1,M2
(Flow cytometry )
Test ControlTest Control
1st signal P. gingivalis LPS
Day 1 2nd signal Cholesterol crystals
Day 0
No treatment
y gCholesterol crystals
Day 2 culture supernatant was collected for IL-1β analysis.
E.Coli LPS as 1st signalPositivecontrol ATP as 2nd signal
Cholesterol crystalsCholesterol crystals
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Monocyte and Macrophage Phenotypes
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Cytokine production from M1 macrophageCytokine production from M1 macrophage
E coli LPS primed P gingivalis LPS primed
M1 macrophageE. coli LPS primed P. gingivalis LPS primed
g/m
l)
175020002250 * *
IL-1β
(pg
200300400500600
1500
* * * *
* **
*
+ ATP
+ CC 2000 ug/ml
+ CC 1000 ug/ml0
100
Control
LPS alone
+ CC 500 ug/ml+ ATP
+ CC 2000 ug/ml
+ CC 1000 ug/ml
Control
LPS alone
+ CC 500 ug/ml
+ C+ C
CC = Cholesterol crystals (Acetone preparation)
+ + C+ C+
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Cytokine production from M2 macrophageCytokine production from M2 macrophage
M2 hE. coli LPS primed P. gingivalis LPS primed
M2 macrophage
2250 *
pg/m
l)
30012501500175020002250
*
*
IL-1β
(p
100
200
300
** *
*
**
*
*
+ ATP
+ CC 2000 ug/ml
+ CC 1000 ug/ml0
Control
LPS alone
+ CC 500 ug/ml+ ATP
+ CC 2000 ug/ml
+ CC 1000 ug/mlContro
l
LPS alone
+ CC 500 ug/ml
+ +
CC = Cholesterol crystals (Acetone preparation)
+ C+ C+
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ConclusionConclusion
“P. gingivalis LPS and cholesterol crystals induced dose-dependent g g y p
IL-1ß secretion from both M1 macrophage and M2 macrophage
via NLRP3 inflammasome activation and no significant differences
in IL-1ß production at each concentration of cholesterol crystals
were observed between two types of macrophages “
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AcknowledgementAcknowledgement
• Associate Professor Dr. Rangsini Mahanonda
• Dr. Sathit Pichyangkuly g
• Dr. Chantrakorn Champaiboon
• Mr. Noppadol Sa-Ard-Iam
• Ms. Pimprapa RerkyenMs. Pimprapa Rerkyen
• NRCT’s research fund
• Staff of Periodontology Department, Chulalongkorn University
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