Animals ; Apolipoproteins E ; deficiency ; genetics ; Blood Vessels ; anatomy & histology ; Fluorescence ; Lipofuscin ; Mice ; Mice, Knockout

OBJECTIVETo explore if PPARgamma agonist rosiglitazone could enhance the anti-atherosclerotic effects of mouse peroxisome proliferator-activated receptor gamma1 (PPARgamma1) gene transfer in apolipoprotein-knock out mice.

METHODSAdult ApoE-knock out mice were fed a Western-diet for 20-weeks and then injected with PBS, Ad. PPARgamma1 (5 x 10(8)pfu) or Ad. GFP (5 x 10(8)pfu) via jugular vein. Another group of mice were intervened with rosiglitazone (dissolved in 0.5% cellulose acetate, 4 mg.kg(-1).d(-1), per gavage) 1 week before Ad. PPARgamma1 injection (n = 10, each group). Two weeks later, the lipid core and plaque composition were characterized with oil red O staining and Movat method respectively. The expression of PPARgamma, SM-actin, MOMA-2, MMP-9/TIMP-1, CD40/CD40L and TF antigens in aortic roots and plaques among four groups were compared semi-quantitatively using immunohistochemical technology.

RESULTSAll parameters were similar between AdGFP and PBS groups (P > 0.05). The area of plaque were significantly decreased and oil red O staining area significantly increased in AdPPARgamma1 [(0.86 +/- 0.12) mm(2), (150 +/- 35) x 10(3) microm(2)] and AdPPARgamma1 + RO [(0.79 +/- 0.15) mm(2), (270 +/- 49) x 10(3) microm(2)] treated mice compared with AdGFP group [(0.98 +/- 0.17) mm(2), (80 +/- 21) x 10(3) microm(2)] all P < 0.05. Elastic fiber, collagen and proteoglycan in plaques were also significantly increased in AdPPARgamma1 and AdPPARgamma1 + RO groups. Upregulation of PPARgamma, SM-actin, TIMP-1 antigen activity and downregulation of MOMA-2, MMP-9, CD40/CD40L and TF antigen activity in AdPPARgamma1 and most significantly in AdPPARgamma1 + RO group were observed (P < 0.05).

CONCLUSIONAnti-atherosclerotic effects of PPARgamma1 gene transfer in ApoE-knock out mice could be enhanced by PPARgamma agonist rosiglitazone.

Animals ; Apolipoproteins E ; deficiency ; genetics ; Atherosclerosis ; genetics ; Gene Transfer Techniques ; Male ; Mice ; Mice, Knockout ; PPAR gamma ; agonists ; genetics ; metabolism ; Peroxisome Proliferator-Activated Receptors ; metabolism ; Thiazolidinediones ; pharmacology ; Transfection

OBJECTIVETo study the relationship between the polymorphism of ApoE gene and TCM syndrome type of primary hyperlipemia.

METHODSApoE genotype of 102 patients with hyperlipemia was detected by gene PCR sequencing.

RESULTSA total of five genotypes were detectable, they were E2/2, E3/3, E4/4, E2/3 and E3/4. The frequency of E3/4 + E4/4 and epsilon4 allelotype detected in the patients of Gan-Shen Yin deficiency syndrome type were significantly higher than those in patients of Pi-Shen Yang-deficiency type or of phlegm stagnation type (P < 0.05, P < 0.01), and which in patients of Qi-stagnation caused blood stasis type were significantly higher than those in patients of phlegm stagnation type ( P < 0.05).

CONCLUSIONPolymorphism of ApoE gene is related in a certain degree to TCM syndrome type of primary hyperlipemia.

Adult ; Aged ; Aged, 80 and over ; Apolipoproteins E ; genetics ; Diagnosis, Differential ; Female ; Genotype ; Humans ; Hyperlipidemias ; diagnosis ; genetics ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Polymorphism, Genetic ; Yin Deficiency ; genetics

BACKGROUNDAtherosclerosis is the primary cause of cardiovascular disease, carotid artery disease, and peripheral vascular disease. However, it is hard to obtain human arterial tissue at different stages of atherosclerosis for a systematic study. The ApoE-deficient (ApoE(-/-)) mice predictably develop spontaneous atherosclerotic plaques with numerous features similar to the human lesions and contain nearly the entire spectrum of lesions observed during atherogenesis in humans. MicroRNA expression profiles at different stages of atherosclerosis in ApoE-deficient mice were screened to find out the differentially expressed microRNAs.

METHODSApoE-deficient mice were euthanized at 4, 8, and 20 weeks of age and divided into three groups according to the three time points, including groups A4 (fed a Western-type diet for 0 week), A8 (fed a Western-type diet for 4 weeks), and A20 (fed a Western-type diet for 16 weeks). Atherosclerotic lesions were analyzed. Fifteen aortas were collected and combined into three pools (five aortas in one pool) in each group. MicroRNA microarray analysis was replicated thrice in each group. The threshold of fold change ≥ 2.0 was used to screen up or down-regulated microRNAs. Differentially expressed microRNAs were subsequently verified with quantitative real-time polymerase chain reaction. Those increasingly up or down-regulated microRNAs during the progression of atherosclerosis were selected.

RESULTSAtherosclerotic lesions first appeared in the aortic arch in group A8. Severe atherosclerotic lesions were observed in group A20. In group A8, seven MicroRNAs were up-regulated while two were down-regulated. In group A20, 15 microRNAs were up-regulated while two were down-regulated. miR-34a-5p and miR-497-5p were increasingly up-regulated, while miR-434-3p was progressively down-regulated when atherosclerosis progressed.

CONCLUSIONSIn this study, we described that microRNAs are differentially expressed at different stages of atherosclerosis in ApoE-deficient mice. Those increasingly up or down-regulated microRNAs during the progression of atherosclerosis may play an important role in the pathogenesis of atherosclerosis and provide us opportunities for investigating atherosclerosis from early to advanced stages.

Animals ; Apolipoproteins E ; deficiency ; genetics ; Atherosclerosis ; genetics ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; MicroRNAs ; genetics ; Real-Time Polymerase Chain Reaction

Animals ; Apolipoproteins E ; deficiency ; Female ; Hyperlipoproteinemia Type III ; blood ; metabolism ; Lipid Metabolism, Inborn Errors ; blood ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Receptors, Calcitriol ; metabolism ; Vitamin D ; blood

OBJECTIVEThis study assessed cardiac function changes post embryonic stem cells (ESCs) perfusion at different concentrations in the isolated apolipoprotein-E gene deficiency (apo E-/-) and wild type (WT) hearts.

METHODSapo E-/- and WT mice hearts were isolated and retrogradely perfused (Langendorff model) and ESCs were infused with different concentrations (Low dose group: 1.0 x 10(6) cells, high dose group: 2.5 x 10(6) cells). Hemodynamic parameters including coronary flow (CF), heart rate (HR), dp/dtmax, dp/dtmin, left ventricular end diastolic pressure (LVEDP), were recorded after stabilization period and at before, 5 min, 15 min and 30 min after cell perfusions. The number of cells in the transudate was counted.

RESULTSCardiac function parameters were similar before cell perfusion in apo E-/- and WT hearts. Cardiac function was significantly impaired after low dose cell perfusion in apo E-/- hearts while remained unchanged in WT hearts with the exception of lowered HR. Cardiac function was also significantly impaired after high dose cell perfusion in both apo E-/- and WT hearts, especially in apo E-/- murine hearts. Most of the cells perfused into the heart were expelled after 30 min (63.2% - 77.0%).

CONCLUSIONESCs perfusion into an isolated heart, especially in the atherosclerosis-prone hearts, in the Langed off model impaired cardiac function in a concentration-dependent manner.

Animals ; Apolipoproteins E ; deficiency ; genetics ; Disease Models, Animal ; Heart ; physiopathology ; In Vitro Techniques ; Mice ; Mice, Knockout ; Myocardial Reperfusion Injury ; etiology ; Stem Cells

OBJECTIVETo clarify the differential expression of the genes related to the lipid metabolism in the early stage of atherosclerosis in the young LDLR-/- mice of different ages.

METHODSA RT-PCR assay was used to analyse the gene expression patterns in the livers of LDLR-/- mice and wild type (WT) mice from 14 to 90 days. The characteristics of early lipid deposition in intima were evaluated using biochemical and pathological techniques.

RESULTSIn LDLR-/- mice, when compared to WT mice, the mRNA level of the apolipoprotein A IV (apoA IV), fatty acid translocase (Fat/CD36) and carnitine palmitoyl transferase I (CPT I) changed prominently at the age of 14-days (P < 0.05). At 30 days, the mRNA level of apolipoprotein A I (apoA I) was up regulated, but apolipoprotein F (apoF), CD36 and CPT I were down regulated (P < 0.05). At 60 days, the mRNA levels of apoA I, CPT I and liver X receptor alpha (LXRalpha) were up regulated, but apoA IV was down regulated (P < 0.05). At 90 days, the level of the apoA I was higher, but the expression of the apoA IV, apoF and acyl-coenzymeA oxidase 1 (ACOX1) were down regulated (P < 0.05), whereas the expression of apolipoprotein A V (apoA V), apolipoprotein E (apoE), peroxidase proliferator-activated receptor alpha (PPARalpha) and angiopoietin-like protein 3 (angptl 3) had no significant changes (P > 0.05). The serum levels of TC (P < 0.05), TG (P < 0.05) and LDLC (P < 0.05) in LDLR-/- mice were significantly higher than those in wild type mice with the same age.

CONCLUSIONSThe mRNA levels of the apoA I, apoA IV, apoF, FAT/CD36, CPT I, ACOX1 and LXRalpha of the LDLR-/- mice were significantly changed compared to the WT mice. The genes may be of some relevance to the complicated lipid metabolism network, and have effect in the early stage of atherogenesis.

Animals ; Apolipoprotein A-I ; genetics ; metabolism ; Apolipoproteins A ; genetics ; metabolism ; Apolipoproteins E ; genetics ; metabolism ; Gene Expression ; Lipid Metabolism ; Liver ; metabolism ; Liver X Receptors ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Orphan Nuclear Receptors ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Receptors, LDL ; deficiency

OBJECTIVETo explore the relationship between the expression characteristics of lipid metabolism-related genes in the liver and early atherosclerotic lesions in apolipoprotein E and low density lipoprotein receptor gene double knockout (apoE(-/-)/LDLR(-/-)) mice.

METHODSRT-PCR was used to detect the differential expression of lipid metabolism-related genes in the liver of apoE(-/-)/LDLR(-/-) and wild type (WT) mice. Serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) level as well as aortic morphology were also analyzed.

RESULTSAmong the 11 lipid metabolism-related genes, apolipoprotein B100 (apoB100) mRNA levels were significantly higher in apoE(-/-)/LDLR(-/-)mice compared with WT mice. At 14 days, 1, 2 and 3 months of age, the level of mRNA expression were 1.55, 1.47, 1.50 and 2.42 folds of those of the age matched WT mice respectively. The fatty acid transporter (FAT/CD36) mRNA expression levels were higher in 14-day and 3-month old mice at 1.30 and 1.35 folds of those of the age matched WT mice, respectively. Apolipoprotein A IV (apoA IV) and Apolipoprotein AV (apoAV) mRNA levels were significantly down-regulated (0.89 fold decrease in 14-day, and 0.90 folds decrease in 3-month, respectively). The mRNA expression levels of apolipoprotein AI (apo AI), apolipoprotein F (apo F), peroxidase proliferator-activated receptor alpha (PPAR-alpha), liver X receptor alpha (LXRalpha), angiopoietin-like protein 3 (ANGPTL3), acyl-coenzymeA oxidase 1 (ACOX1) and carnitine palmitoyl transferase 1 (CPT1) had no significant changes. Serum TC, TG and LDL-C were higher than those of age matched WT mice at 7, 2 and 30 folds, respectively. Furthermore, apoE(-/-)/LDLR(-/-) mice demonstrated typical early atherosclerotic lesions at sinus and root regions of aorta in an age dependent manner.

CONCLUSIONAlterations of the expression of lipid metabolism-related genes in liver play important roles in the development of AS in the apoE(-/-)/LDLR(-/-) mice at early ages.

Animals ; Aorta ; pathology ; Apolipoprotein A-V ; Apolipoprotein B-100 ; biosynthesis ; genetics ; Apolipoproteins ; biosynthesis ; genetics ; Apolipoproteins A ; biosynthesis ; genetics ; Apolipoproteins E ; deficiency ; Atherosclerosis ; etiology ; metabolism ; pathology ; CD36 Antigens ; biosynthesis ; genetics ; Gene Expression ; Lipid Metabolism ; Liver ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; RNA, Messenger ; metabolism ; Receptors, LDL ; deficiency

The aim of this study was to investigate the expression of circulating microRNAs (miRNAs) in apolipoprotein E (apoE) knockout mice (apoE-/-) and to validate the role of these miRNAs in human coronary artery disease (CAD). Pooled plasma from 10 apoE-/- mice and 10 healthy C57BL/6 (B6) mice was used to perform the microarray analysis. The results showed that miR-34a, miR-21, miR-23a, miR-30a and miR-106b were differentially expressed in apoE-/- mice, and these expression changes were confirmed by real-time quantitative reverse-transcription PCR. Then, miR-34a, miR-21, miR-23a, miR-30a and miR-106b were detected in the plasma of 32 patients with CAD and of 20 healthy controls. Only miR-34a, miR-21 and miR-23a were significantly differentially expressed in the plasma of CAD patients (all P<0.01). In conclusion, miR-34a, miR-21 and miR-23a were elevated in CAD patients, which means that these miRNAs might serve as biomarkers of CAD development and progression.
Aged ; Animals ; Apolipoproteins E/deficiency ; Biomarkers ; Case-Control Studies ; Coronary Artery Disease/*genetics ; Disease Models, Animal ; Gene Expression Profiling ; Gene Expression Regulation ; Humans ; Male ; Mice ; Mice, Knockout ; MicroRNAs/*genetics ; Middle Aged ; Pilot Projects ; Reproducibility of Results ; Risk Factors

OBJECTIVETo study the effect of betaine on the formation of atherosclerotic plaque in apolipoprotein E (ApoE)-deficient mice and explore its anti-inflammatory mechanism.

METHODSSeven-week-old ApoE-deficient mice (C57BL/6J background) were divided into four groups randomly based on body weight: model group and three betaine groups. Wild-type mice with the same age and genetic background were used as control group. The control group and model group were fed AIN-93G diet. Three betaine groups were fed AIN-93G diet supplemented with 1, 2, 4 g betaine/100 g diet, respectively. Serum tumor necrosis factor-alpha (TNF-alpha), monocyte chemoattractant protein-1, lipid levels and methylation status of TNF-alpha promotor in aorta were determined at 0, 7 and 14 weeks. The percentage of aorta sinus plaque to lumen area was measured at 14-week.

RESULTSThe percentage of aorta sinus plaque to lumen area of 1% and 2% betaine groups were (11.43+/-2.65)% and (12.09+/-3.07)%, respectively, which were 41% and 33% smaller than that of the model group (t=3.117, 3.010, respectively, and P<0.01). Serum TNF-alpha level of three betaine groups were (56.33+/-3.86), (63.04+/-4.67) and (65.52+/-3.97) pg/ml, respectively, which were lower than that of the model group (79.40+/-4.68) pg/ml (t=9.270, 6.571 and 5.576, respectively, P<0.001), but there was no significant difference in the methylation status of TNF-alpha promotor among all five groups.

CONCLUSIONBetaine could inhibit the development of atherosclerosis via anti-inflammation.

Animals ; Apolipoproteins E ; deficiency ; genetics ; Atherosclerosis ; blood ; drug therapy ; Betaine ; pharmacology ; therapeutic use ; Chemokine CCL2 ; metabolism ; Dietary Supplements ; Disease Models, Animal ; Male ; Mice ; Mice, Inbred C57BL ; Tumor Necrosis Factor-alpha ; metabolism