Biomedical subjects

A Moriguchi

Publications and source records attributed to A Moriguchi.

At least 55 records · Page 3Linked to original sources

PubMed1998

Oligonucleotide-based gene therapy for cardiovascular disease.

Gene therapy is emerging as a potential strategy for the treatment of cardiovascular disease such as restenosis after angioplasty, vascular bypass graft occlusion, transplant coronary vasculopathy, homozygous familial hypercholesterolemia and cystic fibrosis, for which no known effective therapy exists. Gene therapy requires efficient in vivo gene transfer technology. During the past decade, many gene transfer methods including viral transfer techniques have been developed, and some are being applied clinically in human gene therapy studies. Molecular biology and pathophysiology of the cardiovascular system have started to emerge, and the time is ripe for the introduction of gene therapy to the management of cardiovascular disorders. In this review, we have focused on the future potential of oligonucleotide-based gene therapy for the treatment of cardiovascular disease.

Cardiovascular Diseases↗

PubMed1998

Contribution of a vascular modulator, hepatocyte growth factor (HGF), to the pathogenesis of cardiovascular disease.

HGF is a mesenchyme-derived pleiotropic factor which regulates cell growth, cell motility, and morphogenesis of various types of cells, and is thus considered a humoral mediator of epithelial-mesenchymal interactions responsible for morphogenic tissue interactions during embryonic development and organogenesis. Although HGF is originally identified as a most potent mitogen for hepatocytes, HGF is also belonged to a member of endothelium-specific growth factors. Since endothelial cells are known to secrete various anti-proliferative and vasodilating factors, an agent that promotes seeding or regeneration of endothelium may have potential therapeutic value against vascular smooth muscle cell proliferation. The mitogenic action of HGF on human endothelial cells was most potent among growth factors. Moreover, the presence of local HGF system (HGF and its specific receptor, c-met) was observed in vascular cells and cardiac myocytes in vitro as well as in vivo. Production of local HGF production in vascular cells was regulated by various cytokines including transforming growth factor (TGF)-beta and Ang II. Furthermore, HGF may be therapeutic growth factors for the treatment of restenosis after angioplasty and arteriosclerosis oblerance, etc., as gene therapy. On the other hand, serum HGF concentration was significantly correlated with blood pressure. These results suggest that HGF secretion might be elevated in response to high blood pressure as a counter-system against endothelial dysfunction, and may be considered as an index of severity of hypertension. In this review, we discussed the potential role of HGF in cardiovascular disease.

Cardiovascular Diseases↗

PubMed1998

Effect of impression material on surface reactive layer when casting pure titanium in phosphate investment.

We evaluated the effect of impression materials used in preparation of pure titanium castings on the surface reactive layer. Surface roughness of the refractory models before and after firing was smaller when silicone rather than agar impression material was used. The surface roughness of castings prepared with T-invest varied little with the impression material. However, the surface roughness of the castings prepared with CD Titaninvest was less when silicone impression material was used. Surface hardness of the castings was slightly greater when agar impression material was used, and metallic texture analysis of the surface of the castings showed a chill layer and a columnar crystal layer extending from the surface toward the interior. A relatively non-corroded white layer and a markedly corroded black layer were observed in the chill layer, and their thickness was smaller when silicone impression material was used. Use of the Electron Probe Micro Analyzer (EPMA) to determine distribution of various elements in the superficial layer of the casting plates showed that the reactive layer contained less P and Si when silicone impression material was used rather than agar. NH4H2 PO4, which is a component of the bonding material in the investment, was present at a high concentration in the superficial layer of the agar impression material. This shows the importance of preparing refractory models with a non-water-absorbing impression material to obtain pure titanium casting plates with a smaller reactive layer.

Agar↗

PubMed1997

Donor-specific tolerance by perioperative intrathymic injection of bone marrow cells in the rat cardiac allograft model: use of FK506 can shorten the necessary duration of pretransplant intrathymic conditioning.

BACKGROUND: Many strategies of tolerance induction by intrathymic (IT) injection of donor alloantigens have been reported to date; however, the timing of IT injection is usually 1-3 weeks before transplantation. METHODS: To apply IT injection to cadaveric organ transplantation, 1 x 10(8) fully allogeneic bone marrow cells (BMC) of Buffalo (BUF; RT1b) rats were intrathymically injected into Wistar Furth (WF; RT1u) rats at the time of BUF cardiac allografting with short-course therapy of antilymphocyte serum (ALS) and FK506 in our experimental model. RESULTS: Allogeneic IT injection of BUF BMC with ALS and FK506 indefinitely prolonged graft survival (mean survival time > 210 days) in all WF rats. On day 130 after grafting, tolerant WF rats accepted donor BUF skin grafts (> 120 days) but not third-party Lewis skin grafts. In control groups, syngeneic IT injection of WF BMC or intravenous injection of donor BUF BMC in combination with ALS/FK506 therapy failed to induce tolerance. In vivo testing was performed during induction (1 month) or during maintenance (6 months of tolerance. In the mixed lymphocyte reaction (MLR), spleen T cells of tolerant rats at 1 month after grafting displayed hyporesponsiveness after stimulation with donor cells. The addition of interleukin (IL)-2 to MLR culture did not restore T-cell responsiveness. Tolerant rats had a significantly decreased frequency of T cytotoxic cell precursors (fTcp) of 1:4,926, and frequency of IL-2-producing T helper cell precursors (fThp) of 1:23,925, compared with naive rats (1: 2,158 and 1:4,266, respectively). By 6 months after grafting, however, the anti-donor MLR proliferative responses of tolerant rats had been restored to the levels of naive splenic T cells. These tolerant rats displayed restoration of the (fTcp) of 1:2,842 and of the (fThp) of 1:5,630, which were comparable frequencies of naive rats. Suppressor T cells did not contribute in this model. In cardiac grafts of tolerant rats induced by IT injection, expression of both Th1 (interferon-gamma and IL-2) and Th2 (IL-4 and IL-10) cytokines was detected in the early phase; thereafter, expression was completely inhibited, except for interferon-gamma in the chronic phase. CONCLUSIONS: Perfect donor-specific tolerance was obtained by IT injection of donor BMC at the time of transplantation, while alloimmune responses were maintained at levels similar to those of naive rats.

Adoptive Transfer↗

PubMed1997

In vivo transfer efficiency of antisense oligonucleotides into the myocardium using HVJ-liposome method.

Although antisense strategy has been at the center of interest in gene therapy, little is known about application of this strategy to cardiac diseases because of the lack of a suitable delivery method into the heart. Therefore, we compared the transfection efficiency of antisense oligodeoxynucleotides (ODN) using HVJ-liposome method and direct transfer in in vivo transfer into heart. To investigate the cellular fate and localization of ODN, transfer of "naked" FITC-labeled antisense ODN or ODN enwrapped in HVJ-liposome complex were examined. Following in vivo transfer using direct injection as well as in vitro transfer, fluorescence rapidly disappeared within 1 day, whereas transfer by HVJ-liposome method resulted in sustained fluorescence localized in the nucleus for at least 1 week. Measurement of fluorescence also demonstrated a significantly higher level in myocardium transfected by HVJ-liposome method than direct transfer. The present study demonstrated that HVJ-liposome method is more efficacious for ODN delivery by prolongation of half-life of ODN, suggesting its usefulness for gene therapy in cardiac diseases.

Animals↗

PubMed1997

Efficient in vivo gene transfer into the heart in the rat myocardial infarction model using the HVJ (Hemagglutinating Virus of Japan)--liposome method.

The lack of efficient treatment for myocardial infarction remains an unresolved problem in the field of cardiovascular disease. Gene therapy may be a potential therapeutic strategy for the treatment of myocardial infarction. However, current methods of in vivo gene transfer into the heart are limited by their low efficiency and/or potential toxicity. In the present study, we developed an efficient technique of gene transfer into the intact heart in vivo using the Sendai virus (HVJ: Hemagglutinating Virus of Japan)--liposome method. We used the beta-galactosidase gene, luciferase gene and human angiotensin converting enzyme (ACE) gene as markers. In vivo gene transfer into the rat heart was performed as follows: (1) direct injection into the rat heart, (2) incubation within the pericardium, and (3) infusion into a coronary artery. Direct injection of the HVJ-liposome complex containing the beta-galactosidase vector into the rat heart resulted in limited staining of beta-galactosidase 3 days after transfection. To compare transfection efficiency between "naked" plasmid DNA transfection and the HVJ-liposome method, we also transfected the luciferase reporter gene into the heart. Luciferase activity was significantly higher in hearts transfected by the HVJ-liposome method than that in hearts transfected by direct "naked" plasmid transfection (P < 0.01). To confirm the successful gene in the protein level, we measured ACE activity in the hearts. Cardiac ACE activity was significantly increased in hearts transfected with human ACE gene as compared to hearts transfected with control vector (P < 0.01). On the other hand, incubation of HVJ-liposome complex, containing beta-galactosidase vector, within the pericardium resulted in widespread staining of cardiac myocytes and fibroblasts, mainly located in several surface layers beneath the pericardium. More importantly, widespread stained areas of beta-galactosidase were also observed in the middle of the myocardium around the vasa vasorum. We also examined the efficiency of gene transfer by the HVJ-liposome method in a rat myocardial infarction model. In the infarction model, using the pericardium incubation approach, staining for beta-galactosidase was observed in the viable cells around the infarction area. Finally, direct infusion of the HVJ complex, containing the beta-galactosidase vector, into coronary artery also resulted in widespread staining of beta-galactosidase in cardiac myocytes around the microvasculature. Using direct injection, we found significant injury to the myocardium and severe fibrosis at the injection site, whereas no apparent injury was observed using pericardium incubation and coronary infusion. There was no evidence of cytotoxicity or inflammation caused by the HVJ-liposome complex itself. Overall, we have established an efficient in vivo gene transfer method into the heart using the HVJ-liposome method. Direct infusion into the coronary artery resulted in widespread transfection without damaging the myocytes; incubation within the pericardium demonstrated the usefulness of the HVJ-liposome method for studying cardiac function and as a means of gene therapy for cardiovascular diseases.

Animals↗

PubMed1997

Role of hepatocyte growth factor in endothelial regulation: prevention of high D-glucose-induced endothelial cell death by prostaglandins and phosphodiesterase type 3 inhibitor.

Injury of endothelial cells (EC) has been postulated as the initial trigger of the progression of atherosclerosis in patients with diabetes mellitus. We previously reported that decrease in a novel endothelium-specific growth factor, hepatocyte growth factor (HGF), by high D-glucose might be a trigger of endothelial injury. However, the physiological role of the local vascular HGF system has not yet been clarified. To investigate the role of HGF in endothelial injury, we initially examined the effects of HGF on endothelial injury induced by serum deprivation. Decrease in EC number by serum deprivation was significantly attenuated by addition of HGF as well as recombinant basic fibroblast growth factor, whereas vascular endothelial growth factor showed no effect. Apoptotic changes in EC induced by serum deprivation were also significantly attenuated by addition of HGF (p < 0.01). Given the protective action of HGF, we next studied the physiological role of local HGF production in endothelial regulation. We focused on the protective actions of prostaglandin (PG) I2, PGE and a phosphodiesterase type 3 inhibitor (cilostazol) on endothelial injury by high glucose, since these agents are widely used in the treatment of peripheral arterial disease which is frequently observed in diabetic patients. Treatment of human aortic EC with PGE1, PGE2, and a PGI2 analogue (beraprost sodium) as well as cilostazol stimulated EC growth. HGF concentration in conditioned medium from EC treated with PGE1, PGE2 or PGI2 analogue as well as cilostazol was significantly higher than that with vehicle (p < 0.01). Interestingly, treatment with PGI2 analogue or cilostazol attenuated high D-glucose-induced EC death, which was abolished by neutralizing anti-HGF antibody. Moreover, decreased local HGF production by high D-glucose was also significantly attenuated by PGI2 analogue or cilostazol. Finally, we tested the effects of PGE, PGI2 analogue and cilostazol on local HGF production in human aortic vascular smooth muscle cells (VSMC). Although high D-glucose treatment resulted in a significant increase in VSMC number, PGI2 analogue and/or cilostazol treatment had no effects on VSMC growth. However, the decrease in local HGF production by high D-glucose was significantly attenuated by addition of PGI2 analogue or cilostazol. Overall, this study demonstrated that treatment with PGE, PGI2 analogue or cilostazol prevented aortic EC death induced by high D-glucose, probably through the activation of local HGF production. Increased local vascular HGF production by prostaglandins and cilostazol may prevent endothelial injury, potentially resulting in the improvement of peripheral arterial disease.

3',5'-Cyclic-AMP Phosphodiesterases↗

PubMed1997

Iodine-123 metaiodobenzylguanidine myocardial scintigraphy for prediction of response to beta-blocker therapy in patients with dilated cardiomyopathy.

This study was performed to evaluate whether iodine-123 metaiodobenzylguanidine (MIBG) myocardial scintigraphy could predict the response to beta-blocker therapy in patients with nonischemic dilated cardiomyopathy (DCM). Beta-Blocker therapy is effective in some patients with DCM. MIBG myocardial scintigraphy has also been suggested to be useful in evaluating the severity of myocardial damage in DCM. However, no data exist on whether MIBG imaging can be used to predict which patients with DCM will respond to beta-blocker therapy. We prospectively evaluated whether MIBG myocardial imaging was useful in predicting responses to beta-blocker therapy in patients with DCM. MIBG imaging was performed in 45 patients with DCM (35 men, 10 women, aged 13 to 68 years) before the start of bisoprolol. The heart to mediastinum (H/M) MIBG uptake ratio was evaluated on initial and delayed images, and the percent washout rate of myocardial MIBG was also obtained from these data. Of the 45 patients, 30 (67%) responded to beta-blocker therapy, whereas 2 were resistant and 13 showed progression of heart failure or died of heart failure. By logistic regression analysis, the H/M uptake ratio on delayed images was seen to be a good predictor of the response to beta-blocker therapy with a threshold of 1.7 (sensitivity = 91%, specificity = 92%, accuracy = 91%, positive and negative predictive value = 97% and 80%, respectively). These results indicate that an H/M ratio > 1.7 on the delayed MIBG myocardial scintigraphic images provides a useful indication of whether patients with DCM will respond to beta-blocker therapy.

3-Iodobenzylguanidine↗

PubMed1997

Left ventricular Doppler filling pattern in dilated cardiomyopathy: relation to hemodynamics and left atrial function.

This study attempted to examine the relation of left ventricular filling patterns to hemodynamic status and left atrial function in dilated cardiomyopathy. Transesophageal echocardiography and cardiac catheterization were performed in 41 patients with dilated cardiomyopathy (six with an ischemic origin). Transmitral, left atrial appendage, and pulmonary venous flow velocities were recorded with the pulsed Doppler method. Left atrial systolic function was assessed by the peak velocity of the left atrial appendage emptying wave and pulmonary venous flow reversal during atrial systole. Patients were classified into three groups according to their left ventricular filling patterns. Group 1 showed an abnormal relaxation pattern (E wave/A wave ratio <1, n = 17), group 2 had a normal or pseudonormal pattern (1 < or = E/A < 2, n = 11), and group 3 had a restrictive pattern (E/A > or = 2, n = 13). No differences were found among the groups with regard to age, gender, heart rate, and M-mode echocardiographic indices of left ventricular function. Compared with patients in group 1, those in groups 2 and 3 had more symptoms (New York Heart Association functional class III or IV) and had higher left ventricular filling pressures. The sensitivity of an E/A ratio > or = 1 for predicting a pulmonary capillary wedge pressure > or = 15 mm Hg was 75% and the specificity was 94%. Despite a similar increase of filling pressures, group 3 patients had a lower left atrial appendage emptying velocity, pulmonary venous flow reversal velocity, and mitral A velocity than did group 2 patients. The sensitivity and specificity of an E/A ratio > or = 22 for detecting left atrial dysfunction (left atrial appendage emptying velocity < or = 40 cm/sec) was 85% and 86%, respectively. In conclusion, among patients with dilated cardiomyopathy, those who had a restrictive or pseudonormal filling pattern were in a higher functional class and had higher filling pressures. Further studies are needed to determine the therapeutic and prognostic significance of left atrial dysfunction, which was common in patients with a restrictive pattern.

Adult↗

PubMed1997

In vivo transfection of cis element "decoy" against nuclear factor-kappaB binding site prevents myocardial infarction.

The transcriptional factor nuclear factor-kappaB (NFkappaB) plays a pivotal role in the coordinated transactivation of cytokine and adhesion molecule genes that might be involved in myocardial damage after ischemia and reperfusion. Therefore, we hypothesized that synthetic double-stranded DNA with high affinity for NFkappaB could be introduced in vivo as "decoy" cis elements to bind the transcriptional factor and to block the activation of genes mediating myocardial infarction, thus providing effective therapy for myocardial infarction. Treatment before and after infarction by transfection of NFkappaB decoy, but not scrambled decoy, oligodeoxynucleotides before coronary artery occlusion or immediately after reperfusion had a significant inhibitory effect on the area of infarction. Here, we report the first successful in vivo transfer of NFkappaB decoy oligodeoxynucleotides to reduce the extent of myocardial infarction following reperfusion, providing a new therapeutic strategy for myocardial infarction.

Animals↗

PubMed1997

Survival of grafts of genetically modified cardiac myocytes transfected with FITC-labeled oligodeoxynucleotides and the beta-galactosidase gene in the noninfarcted area, but not the myocardial infarcted area.

Since one of the attractions of gene therapy in the heart is the implantation of genetically modified cultured cells, we employed genetically modified myocytes transfected with FITC-labeled oligodeoxynucleotides (ODN) and beta-galactosidase gene in this study, to investigate the cellular localization and fate of grafted myocytes in the heart. In addition, we examined the feasibility of myocytes grafting into a myocardial infarction model. Delivery of FITC-labeled ODN with the HVJ-liposome method resulted in sustained fluorescence as compared to direct transfer of 'naked' ODN. Interestingly, implantation of cardiac myocytes transfected with FITC-labeled ODN ex vivo by the HVJ-liposome method resulted in sustained fluorescence for at least 1 week in the noninfarcted area, whereas little fluorescence was detected in the area of infarction. This observation was confirmed by measurement of fluorescence, which showed significantly higher levels in the noninfarcted area than the infarcted area. Positive staining for beta-galactosidase protein was also clearly observed 7 days after grafting of transfection of the beta-galactosidase gene, while no staining was detected in grafted myocytes in control rats. Survival of implanted genetically modified cardiac myocytes in the noninfarcted, but not infarcted area, provides new information for the local delivery of recombinant molecules to the heart using gene therapy.

Animals↗

PubMed1997

Renal effects of an angiotensin II antagonist in stroke-prone spontaneously hypertensive rat.

We evaluated the renal effects of the new angiotensin II type 1 (AT ) receptor antagonist, HR 720, in the stroke-prone spontaneously hypertensive rat. Rats were treated with either vehicle, HR 720, MK-954 (a selective AT1 receptor antagonist) or enalapril for 6 weeks. Blood pressure was decreased to a similar extent by HR 720, MK-954 and enalapril (203 +/- 4, 202 +/- 5 and 190 +/- 4 vs. 247 +/- 4 mm Hg for control). Urinary protein secretion was also decreased (5.2 +/- 0.3, 5.3 +/- 0.2 and 5.5 +/- 0.6 vs. 25.2 +/- 4.6 mg/100g/24h). The glomerular hypertensive change was improved in each drug-treated group (2.0 +/- 0.2, 3.3 +/- 0.3 and 1.6 +/- 0.1 vs. 17.6 +/- 1.5%; p < 0.0001). These results show that, in addition to its antihypertensive effect, HR 720 has a beneficial effect on renal function.

Angiotensin I↗

PubMed1997

Role of angiotensin II in the regulation of a novel vascular modulator, hepatocyte growth factor (HGF), in experimental hypertensive rats.

Hepatocyte growth factor (HGF) is a mesenchyme-derived pleiotropic factor that regulates cell growth, cell motility, and morphogenesis of various types of cells, and is thus considered a humoral mediator of epithelial-mesenchymal interactions responsible for morphogenic tissue interactions. We have previously reported that HGF is a novel member of endothelium-specific growth factors whose serum concentration is positively associated with blood pressure in humans. Therefore, we speculated that serum HGF secretion might be elevated in response to high blood pressure as a counter-system against endothelial dysfunction. However, it is difficult to elucidate the role of circulating and tissue HGFs in human hypertension. To address this issue, we measured circulating and tissue HGF concentrations in spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) at different ages. Serum HGF concentration in SHR was significantly higher than that in WKY at 6, 15, and 25 weeks of age (P<.01). Serum HGF concentration was also significantly positively correlated with blood pressure in SHR (P<.02, r=.455). In contrast, tissue HGF concentrations in heart, aorta, and kidney were significantly decreased in SHR as compared with WKY at 25 weeks of age, when these organs showed hypertrophic changes induced by hypertension (P<.01). Cardiac HGF mRNA was also decreased in SHR as compared with WKY at 25 weeks of age. Moreover, cardiac HGF concentration showed a significant negative correlation with left ventricular (LV) weight (P<.01), whereas serum HGF concentration showed a significant positive correlation with LV weight (P<.05). Interestingly, concentrations of cardiac and vascular angiotensin II, a suppressor of HGF, were increased in SHR as compared with WKY at 25 weeks of age (P<.01). Therefore, we examined the effects of angiotensin blockade on circulating and tissue HGF concentrations, to study the role of angiotensin II in HGF regulation. Administration of an angiotensin-converting enzyme inhibitor (enalapril) and angiotensin II type 1 receptor antagonists (losartan and HR 720) for 6 weeks resulted in a significant increase in cardiac HGF concentration, accompanied by increased cardiac HGF mRNA, and a significant decrease in serum HGF concentration, accompanied by lowered blood pressure and reduced LV weight (P<.01). Here, we demonstrated increased circulating HGF and decreased vascular, cardiac, and renal HGF in SHR as compared with WKY at the maintenance stage of hypertension. Decreased tissue HGF in target organs of hypertension may be due to increased tissue angiotensin II. These results suggest that decreased local HGF production may have an important role in the cardiovascular remodeling of target organs in hypertension, since HGF prevented endothelial injury and promoted angiogenesis. Blockade of angiotensin augmented local decreased cardiovascular HGF in hypertension, potentially resulting in the improvement of endothelial dysfunction.

Aging↗

PubMed1997

Comparison of immediate and long-term outcome of percutaneous transvenous mitral commissurotomy in patients who have and have not undergone previous surgical commissurotomy.

This study compared the immediate and long-term outcome of percutaneous transvenous mitral commissurotomy (PTMC) in patients who had (restenosis group, n = 9) or had not (de novo group, n = 27) previously undergone surgical mitral commissurotomy. The baseline echocardiographic score, which is an index of deformity of the mitral valve apparatus, was significantly higher in the restenosis group than in the de novo group (11 +/- 3 vs 7 +/- 2, p < 0.01), although age, left atrial diameter, and the prevalence of atrial fibrillation were similar. PTMC was performed by the Inoue technique, and was abandoned in 1 patient from the restenosis group because of failed trans-septal puncture. Including this patient, 3 patients (33%) in the restenosis group had a thickened atrial septum compared with only 1 (4%) in the de novo group. One patient in the de novo group developed cardiac tamponade during this procedure. In both groups, the mitral valve area increased significantly, but the success rate of PTMC was lower in the restenosis group (4/9 patients, 44%) than in the de novo group (22/27 patients, 81%) (p < 0.05). Twenty-six patients who had successful PTMC were followed up over 51 +/- 14 months. After 4 years of follow-up. 3 out of 4 patients (75%) in the restenosis group and 3 out of 22 patients (14%) in the de novo group demonstrated echocardiographic restenosis (p < 0.01). Stepwise multivariate analysis revealed that the echocardiographic score was the only significant predictor of both the immediate and long-term outcome. In conclusion, the immediate and long-term outcome of PTMC were worse in patients who had undergone previous surgical mitral commissurotomy than in those who had not. This was mainly attributable to the difference in the severity of the valvular lesions. In addition, our data suggested that a thickened atrial septum, possibly related to surgery as well as chronic rheumatic disease, may affect the performance of PTMC.

Adult↗

PubMed1997

Potential role of an endothelium-specific growth factor, hepatocyte growth factor, on endothelial damage in diabetes.

Endothelial cells are known to secrete various antiproliferative and vasodilating factors. Although injury of endothelial cells has been postulated as an initial trigger of the progression of atherosclerosis in patients with diabetes, the mechanisms of endothelial injury in diabetes are not yet clarified. Therefore, it is important to know the effects of high glucose on the factors that may influence endothelial cell growth. A novel member of endothelium-specific growth factors, hepatocyte growth factor (HGF), is produced in vascular cells. To investigate the effects of high glucose on vascular cells, we examined 1) the effects of high glucose on endothelial cell and vascular smooth muscle cell (VSMC) growth and 2) the effects of high glucose on local HGF production in endothelial cell and VSMC. Treatment of human aortic endothelial cell with a high concentration of D-glucose, but not mannitol and L-glucose, resulted in a significant decrease in cell number. Interestingly, addition of recombinant HGF attenuated high D-glucose-induced endothelial cell death. Therefore, we measured local HGF secretion of endothelial cell. Importantly, local HGF production was significantly decreased by high D-glucose treatment. In contrast, high D-glucose treatment resulted in a significant increase in the number of human aortic VSMCs, whereas local HGF production was significantly decreased in accordance with increase in D-glucose concentration. No significant changes in numbers were observed in VSMC treated with high mannitol and L-glucose. We also studied the mechanisms of local HGF suppression by high D-glucose. High D-glucose treatment stimulated transforming growth factor-beta (TGF-beta) concentration in endothelial cell and VSMC. Decreased local vascular HGF production was abolished by addition of anti-TGF-beta antibody. As TGF-beta inhibited local HGF production in endothelial cell and VSMC, increased TGF-beta induced by high D-glucose may suppress local HGF production. This study demonstrated that high D-glucose induced endothelial cell death, stimulated VSMC growth, and decreased local HGF production through the stimulation of TGF-beta production both in endothelial cell and VSMC. Overall, decrease in a local endothelial stimulant, HGF, by high D-glucose may be a trigger of endothelial injury in diabetes, potentially resulting in the progression of atherosclerosis.

Animals↗

PubMed1997

Potential role of a novel vascular modulator, hepatocyte growth factor (HGF), in cardiovascular disease: characterization and regulation of local HGF system.

Since endothelial cells (EC) are known to secrete various anti-proliferative and vasodilating factors, an agent that promotes seeding or regeneration of EC may have potential therapeutic value against vascular smooth muscle cell (VSMC) proliferation. To seek an endothelium specific growth factor, we have focused on hepatocyte growth factor (HGF). HGF is belonged to a member of endothelium specific growth factors, whose mitogenic action on EC was most potent among growth factors. Moreover, the presence of local HGF system (HGF and its specific receptor, c-met) was observed in EC and VSMC of rat and human in vitro as well as in vivo. Production of local HGF production in vascular cells was regulated by various cytokines including transforming growth factor (TGF)-beta and angiotensin II (Ang II). Furthermore, HGF may be a therapeutic growth factor for the treatment of restenosis after angioplasty and arteriosclerosis obliterance, etc., as gene therapy. From these characteristics of HGF, we hypothesized that HGF might contribute to the protection or repair of vascular endothelial cells. Indeed, serum HGF concentration was significantly correlated with blood pressure, suggesting that HGF secretion might be elevated in response to high blood pressure as a counter-system against endothelial dysfunction. In this review, we discussed that HGF is a member of the endothelium specific growth factors whose serum concentration is significantly associated with blood pressure.

Animals↗

PubMed1997

Serum deprivation-induced apoptosis accompanied by up-regulation of p53 and bax in human aortic vascular smooth muscle cells.

Vascular remodeling, which plays an important role in atherosclerosis and hypertension, is determined in large part by the balance between cell growth and cell death by apoptosis. In this study, we studied the apoptosis of human aortic vascular smooth muscle cells (VSMC) induced by serum deprivation. Serum deprivation induced apoptosis of VSMC in a time-dependent manner. Serum deprivation resulted in the up-regulation of p53 protein, compared to treatment with 10% serum (P < 0.01), suggesting that apoptosis induced by serum deprivation may be due to the up-regulation of p53. Of importance, the protein of bax, a promoter of apoptosis, was significantly increased in VSMC treated by serum deprivation compared to treatment with 10% serum (P < 0.01). Overall, these findings demonstrated that serum deprivation induced apoptosis in human aortic VSMC, accompanied by the induction of p53 and bax, suggesting that apoptosis induced by serum deprivation may be mediated by the p53-bax pathway.

Apoptosis↗

PubMed1996

Autocrine-paracrine effects of overexpression of hepatocyte growth factor gene on growth of endothelial cells.

Although hepatocyte growth factor (HGF) is synthesized in vascular cells, it is not known whether locally synthesized HGF acts similarly to exogenously added HGF. Therefore, we transfected cultured cells with human HGF vector and examined the effects on growth of vascular cells. Endothelial cells (EC) transfected with HGF vector synthesized and secreted high levels of HGF, and also showed significantly higher number. Addition of conditioned medium from vascular smooth muscle cells (VSMC) or EC transfected with HGF vector to nontransfected EC resulted in a significant increase in cell number, which was abolished by anti-HGF antibody. Co-culture of HGF-transfected VSMC with EC showed that HGF released from VSMC or EC stimulated EC growth. These results demonstrate that endogenously produced HGF by transfection of human HGF vector can exert autocrine and paracrine stimulatory effects on EC growth, but not VSMC growth, suggesting the role of local HGF system in cardiovascular disease.

Animals↗