Distinct phenotypes in severe hypertriglyceridemic mice: spontaneous atherosclerosis, pancrearitis, impaired hemorheology and abnormal neuronal development
刘国庆
北京大学心血管研究所
HyperTG in the world:
Meta, multi-variables, Epidemiological analysis:
HyperTG is an independent risk factor of coronary artery disease
S447X, a naturally occurring C-terminal two amino acid truncation mutation
wild type LPL ----------------------KKSG
S447X LPL ----------------------KK
Incidence: 18.8% of Caucasian, 25.4% Chinese, and 3% of Australian Aborigine
associated with: Low TG, High HDL-C, Low CHD
Complete Rescue of Lipoprotein Lipase-Deficient Mice by Somatic Gene Transfer of the Naturally Occurring LPLS447X Beneficial Mutation. Arterioscl. Thromb. Vasc. Biol. 2005 Jul 7
% Survival rate of LPL-/- pups
Days after Birth
The Unique HyperTG Mouse Models
Adult LPL-/-mice
TC TG
WT 58± 5.8 79±22.0
ApoE0 304± 10.2* 145±51.9 *
LPL0 189± 58.0* 3199±773.2*
ApoE0/LPL0 817±166.7* 5988±857.4*
Plasma Lipids Levels of KO Mice
AAV-Mediated long term LPL gene therapy for murine extreme HTG
LK-/- > 15 m
WT > 15 m
Lesion Quantitation
LK-/- 10m
WT 10m
LK+/- 10m
3 out of 8, 37％
WT
LK+/-
LK-/-
4m
>15m
10m
Pending
LK-/- 4m
WT 4m
LK+/- 4m
（40×）
Macrophage
WT 10M
Oil red O
LK+/- 15 m
LK-/- 15 m
（200×）
VCAM-1
VCAM-1
Oil red O
（200×）
LK+/- 15 m
LK-/- 15 m
4 month
10 month
>15 month
WT
+/-
-/-
WT
WT
-/-
-/-
CM oxidation
15m
10m
4m
Oxidation of chylomicron by ESR analysis
RO production as reflected by
peak Height of ESR graph
4m
10m
15m
0
1
2
3
4
5
6
4 m
10 m
15 m
Product of RO
.
(cm/mg)
Spontaneous atherosclerosis in hypertriglyceridemic murine model on normal chow diet
Alterations of Blood Coagulation
Hemorheological changes in the mice
Hematocrit RBC Electroph. Rate
% of RBC Vol.
wt
E0
*
*
50.0
37.5
25.0
12.5
0
L0
E0/L0
1.00
0.75
0.50
0.25
0
Electrophoresis rate
Alterations of RBC Rheology
Alterations in RBC polarization and microviscosity
Polarization Microviscosity
P value of RBC ghost
wt
E0
*
*
0.20
0.15
0.10
0.05
0
L0
E0/L0
1.00
0.75
0.50
0.25
0
η value of RBC ghost
*
*
*
*
Structural changes in RBC membrane as studied by FTIR: 2. Secondary structure of membrane protein
?-turns α-helix ?-sheet α-helix
A=1684 A=1659 A=1634 A=1545
Absorption value
wt
E0
*
*
2.00
1.50
1.00
0.50
0
L0
E0/L0
*
*
*
*
*
*
*
*
*
*
*
Hemorrheologic abnormalities in mice with severe hypertriglyceridemia and combined hypertriglyceridemia/cholesterolemia
Impact of extreme hypertriglyceridemia on atherogenesis: utilization of ApoE/ LPL double deficient mice
Aortic lesion development at 3, 6 and 9 month compared with ApoE0 and E0/LPL+/-mice;
Local expression of active and inactive hLPL in carotid arteries: identification of lipid deposition;
BMT as tool for evaluation of macrophage-specific deletion of LPL gene;
As model by FeCl injury on carotid artery for evaluation of the LPL’s effect on atherogenesis
ApoE-/- XLPL-/- ApoE-/-
6 months
3 months
ELK -/-
EK -/-
ELK +/-
（40×）
3 M
6 M
9 M
Pending
Pending
Pending
Pending
Pending
Pending
External carotid artery
Common carotid artery
Internal carotid artery
Arterial wall gene transfer to mouse carotid artery
Local expression of active and inactive hLPL in carotid arteries in WT mice: RealTime quantitation of mRNA
1000 bp
500 bp
(-) AP WT 194 P M
1 2 3 4 5 6
β-actin (500bp)
400 bp
LPL (396 bp)
AP LPL-WT LPL194
Ratio of LPL/ actin (X103)
1
2
Ctl. LPL staining
LPL GT. LPL Staining
Ctl. ORO staining
LPL GT. ORO staining
Arterial expression of LPL gene in WT mice:
Promotion of Lipid Deposition
ORO staining
hAP hLPLWT hLPL194
(200x)
(400x)
ApoE-/-
ORO staining
(200x)
(400x)
LPL-/-
Ad-AP
Ad-LPLwt
Ad-LPL194
en face confocal microscopy images of VCAM-1 expression
LPL-/-
Rapid lipid deposition in endothelial-intact carotid artery via arterial expression of either active or inactive lipoprotein lipase in ApoE and LPL deficient mouse models.
Common carotid arteries of apoE-/-, LPL+/- (ELK)mice (A) and apoE-/-, LPL+/+ (EK)mice (B) 8 weeks after ferric chloride injury
neointima formation after carotid artery injury by FeCl3 for 8 weeks
ELK
carotid arter