Advanced atherosclerosis is often associated with dystrophic calcification and remodeling of extracellular matrix of vascular wall. Recently many studies have documented a general relationship between calcification and severity of coronary disease, and discussed the feasibility of electron beam computed tomography for detecting and quantifying the coronary artery calcification in the patients. The present study investigated the expression and the localization of osteopontin, one of noncollagenous bone matrix protein, within the calcified coronary arteries. Autopsy-derived coronary artery specimens were scanned and reconstructed to visualize the pattern of coronary calcification using a novel microscopic computed tomography technique. The localization of the osteopontin were evaluated by immunohistochemial stain with LF7. The present study showed that the pattern of coronary calcification is variable and the expression of osteopontin is localized mainly to calcified lesion. The smooth muscle cells in addition to macrophage expressed osteopontin protein in human coronary atherosclerotic plaques. Soluble osteopontin released near to the sites of vascular calcification may represent an adaptive mechanism aimed at regulating the process of vascular calcification.
Aged ; Calcinosis/metabolism ; Coronary Arteriosclerosis/pathology* ; Coronary Arteriosclerosis/metabolism* ; Coronary Vessels/pathology* ; Coronary Vessels/metabolism ; Coronary Vessels/chemistry* ; Female ; Human ; Immunohistochemistry ; Male ; Middle Age ; Sialoglycoproteins/biosynthesis ; Sialoglycoproteins/analysis*

BACKGROUND: The interindividual variability for the development of collaterals in coronary artery disease is dependent on the hypoxic induction level of VEGF. To determine whether the hypoxic induction of VEGF is controlled by the transcription of HIF-1 (alpha), the VEGF and HIF-1 (alpha) m-RNA levels were correlated to hypoxia in monocytes harvested from patients with coronary artery disease. METHODS: The collateral scoring system used was modified from the TIMI system. The mononuclear cell layer of the patients' blood was cultured in hypoxia (1% O2, 5% CO2, 94%N2) and normoxia (5% CO2, 95% room air) for 17 hours. The VEGF and HIF-1 (alpha) mRNA levels were measured using a RT-PCR technique. We calculated the fold inductions of VEGF, HIF-1 (alpha) mRNA with hypoxia by dividing thehypoxic and the normoxic values. RESULTS: We found significantly higher hypoxic inductions of VEGF m-RNA in patients with collaterals compared to patients with no collaterals. However, there was no difference in the hypoxic inductions of HIF-1 (alpha) between the two groups (VEGF m-RNA mean fold inductions 3.71+/-3.30 versus 1.65+/-0.62, p=0.012, HIF-1 (alpha) mRNA 1.42+/-0.58 versus 1.20+/-0.39, p=0.165). CONCLUSIONS: We concluded that the interindividual variability in the hypoxic inductions of VEGF m-RNA in monocytes in patients is not controlled by the transcriptional levels of HIF-1 (alpha) with hypoxia. These findings suggest that a mechanism such as the post-transcriptional modification of HIF-1 (alpha) is involved in the hypoxic inductions of VEGF.
Vascular Endothelial Growth Factor A/*metabolism ; RNA, Messenger/metabolism ; Monocytes/metabolism ; Middle Aged ; Male ; Hypoxia-Inducible Factor 1, alpha Subunit/*metabolism ; Humans ; Female ; *Coronary Circulation ; Coronary Arteriosclerosis/metabolism ; *Collateral Circulation ; Cells, Cultured ; Anoxia/*metabolism

In this study of the inhibitory effects of angiopeptin and aspirin on the development of accelerated graft atherosclerosis (AGAS), 22 B10.BR mice received intra-abdominal heterotopic heart transplants from B10.A mice, without immunosuppression. Group 1 (n = 5) received no pharmacological intervention, Group 2 (n = 6) was treated with angiopeptin, Group 3 (n = 5) with aspirin, and Group 4 (n = 6) with both. There was no significant difference in the incidence of AGAS among these groups. The magnitude of intimal lesion development showed less narrowing of large vessels (> 100 microns in diameter) in groups 2 and 4--i.e. the groups received angiopeptin (Group 1 = 46.9 +/- 9.3%, Group 2 = 28.5 +/- 9.2%, Group 3 = 44.1 +/- 10.9%, Group 4 = 24.2 +/- 5.9%; p < 0.01). Comparison of the fraction of tropomyosin-positive staining cells in the intima revealed a lesser degree of staining in Group 2 (p < 0.01). No intervention was effective in preventing smooth muscle cell proliferation in the media or inflammatory cell infiltration in the adventitia. In conclusion, our data suggest that angiopeptin is effective in the direct inhibition of intimal smooth muscle cell proliferation in relatively large vessels, whereas aspirin exhibits no inhibitory role in the progression of AGAS. Angiopeptin appears to be a potential therapeutic agent for inhibiting the progression of postoperative AGAS in clinical heart transplantation.
Animal ; Aspirin/pharmacology* ; Cardiovascular Agents/pharmacology* ; Coronary Arteriosclerosis/pathology ; Coronary Arteriosclerosis/immunology* ; Coronary Vessels/pathology ; Coronary Vessels/drug effects ; Heart/drug effects* ; Heart Transplantation/immunology* ; Immunohistochemistry ; Mice ; Mice, Inbred Strains ; Myocardium/pathology ; Myocardium/immunology ; Oligopeptides/pharmacology* ; Somatostatin/pharmacology ; Somatostatin/analogs & derivatives* ; Transplantation, Homologous/immunology ; Tropomyosin/metabolism

The formation of advanced glycation end products (AGEs), in various tissues has been known to enhance immunoinflammatory reactions and local oxidant stresses in long standing diabetes. Recently, AGEs have been reported to play a role in neointimal formation in animal models of arterial injury. We attempted to determine whether the serum levels of AGEs are associated with coronary restenosis in diabetic patients. Blood samples were collected from diabetic patients with coronary artery disease undergoing stent implantation and the serum levels of AGEs were analyzed by the fluorescent intensity method. The development of in-stent restenosis (ISR) was evaluated by a 6-month follow-up coronary angiography. A total of 263 target lesions were evaluated, in 203 patients. The ISR rate in the high-AGE (> 170 U/ml) group (40.1%) was significantly higher than in the low-AGE group (< or =170 U/ml) (19.6%) (p < 0.001). Furthermore, multivariate analysis revealed that a high level of serum AGEs is an independent risk factor for the development of ISR (odds ratio, 2.659; 95% CI, 1.431-4.940; p=0.002). The serum levels of AGEs constitute an excellent predictive factor for ISR, and should be one of the guidelines for medical therapy and interventional strategy to prevent ISR in diabetic patients.
Aged ; Coronary Arteriosclerosis/epidemiology/*metabolism/*therapy ; Coronary Restenosis/epidemiology/*metabolism ; Diabetes Mellitus, Type 2/epidemiology/*metabolism ; Female ; Glycosylation End Products, Advanced/*blood ; Humans ; Male ; Middle Aged ; Research Support, Non-U.S. Gov't ; Risk Factors ; *Stents

Animals ; Arteriosclerosis ; blood ; Blood Glucose ; metabolism ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Coronary Artery Disease ; blood ; Fatty Liver ; blood ; Humans ; Triglycerides ; blood

OBJECTIVETo investigate the effect of chronic enhanced external counterpulastion (EECP) on gene expression profiles of arterial endothelial cells (ECs) of pigs fed with high-cholesterol diet.

METHODSEight male pigs were fed with high-cholesterol diet for 12 weeks to induce arteriosclerosis and subjected to EECP for accumulative 36 h (2 h every other day for 18 sessions). Another 8 pigs on cholesterol-enriched diet and 6 normally fed pigs served as the arteriosclerosis model group and normal control group, respectively, and the high-cholesterol diet was maintained until the end of EECP treatment. The coronary artery was then isolated for transmission electro microscopy, and the abdominal aorta was observed using Sudan III staining. The gene expression profiles in ECs from the thoracic aorta using cDNA microarrays.

RESULTSMacrophages and foam cells were detected beneath the ECs in the coronary artery of pigs in the model group, but not in the other two groups. The ratios of Sudan III-positive area in the celiac aorta were significantly lower in normal control and EECP groups than in the model control group (P<0.05). Compared with the normal control group, the gene expressions of integrins-beta1 and CTGF were up-regulated in the model group. Compared with the model group, the expressions of integrins-beta1, CTGF and VCAM-1 were down-regulated and eNOS up-regulated in EECP group.

CONCLUSIONChronic EECP may reduce endothelial injury, down-regulate the gene expression level of integrin-beta1, CTGF and VCAM-1, lower cholesterol uptake and attenuate arterial endothelial inflammation to protect the pigs fed with high-cholesterol diet from arteriosclerosis.

Animals ; Aorta, Abdominal ; metabolism ; pathology ; Arteriosclerosis ; etiology ; genetics ; pathology ; Coronary Vessels ; metabolism ; pathology ; Counterpulsation ; methods ; Diet, Atherogenic ; Endothelial Cells ; metabolism ; Gene Expression Profiling ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; Swine

We evaluated the risk of coronary-artery disease in patients with chronic renal failure (CRF) by measuring the coronary-artery calcium scores with electron beam CT (EBCT). A total of 81 CRF patients were divided into three groups; pre-dialysis (group I, n=35), hemodialysis (group II, n=31) and peritoneal dialysis (group III, n=15). The several serum biochemical markers and calcium score levels by EBCT were determined. The Ca x P products were significantly higher in groups II (p<0.05) and III (p<0.01) than in group I. The serum calcium levels were significantly higher in group III than in both group I (p<0.01) and II (p<0.05). The serum calcium level in 15 patients with a calcium score > 400 was significantly higher than the 66 patients with a score < or =400 (p<0.01). The calcium score was significantly higher in the 15 patients with cardiovascular complications than in the 66 patients without cardiovascular complications (628.9+/-904.8 vs. 150.4+/-350.9, p<0.01). EBCT seemed to be a good diagnostic tool for evaluating the risk of coronary-artery disease ''noninvasively'' in CRF patients who are at increased risk of cardiovascular morbidity and mortality.
Adolescent ; Adult ; Aged ; Calcinosis/etiology/metabolism/*radiography ; Calcium/blood/*metabolism ; Coronary Arteriosclerosis/etiology/metabolism/radiography ; Coronary Vessels/*metabolism ; Female ; Humans ; Kidney Failure, Chronic/complications/metabolism/*radiography/therapy ; Male ; Middle Aged ; Peritoneal Dialysis ; Renal Dialysis ; Research Support, Non-U.S. Gov't ; Risk Factors ; Tomography, X-Ray Computed

The heart transplantation-associated accelerated graft arteriosclerosis (AGAS) is one of the major causes of cardiac allograft failure. We investigated the early time-course of expresssion patterns of cytokines, transcription factor, and its inhibitor in the intraabdominally transplanted mice hearts that differed only in the D locus of class I histocompatibility antigen. The allograft hearts were harvested at 1-3, 5, 7, 14, 28, and 42 days after the transplantation, and the expressions of NF-kappaB/I-kappaB and cytokines (TNF-alpha , INF-gamma) were examined in these specimens. The expressions of TNF-alpha and INF-gamma were observed on day 1, peaking on day 5 and 7, respectively. Activated NF-kappaB (p65) expression was present on the cytoplasm and perinuclear area in the endothelial cells of coronary arteries on day 1. The peak of translocation of NF-B from cytoplasm to nucleus appeared on day 5 in the endothelial cells, myocytes, and leukocytes within the vessels, and remained elevated until day 42. The I-kappaB expression gradually increased from day 1 until day 5, but a remarkable decrease was detected on day 7. Our data suggest that the increased expressions of NF-kappaB/I-kappaB and cytokines (TNF-alpha, INF-gamma) play an important role in inducing immune responses in the donor allograft heart and hence the blockage of the expressions might be mandatory to avoid a potential graft failure.
Animal ; Chronic Disease ; Coronary Arteriosclerosis/etiology/*metabolism ; Cytokines/*biosynthesis ; *Graft Rejection ; *Heart Transplantation ; Histocompatibility Antigens Class I/analysis ; Intercellular Adhesion Molecule-1/biosynthesis ; Interferon Type II/biosynthesis ; Mice ; NF-kappa B/*biosynthesis ; Transplantation, Homologous ; Tumor Necrosis Factor/biosynthesis ; Vascular Cell Adhesion Molecule-1/biosynthesis