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Ming-hui Zou - One of the best experts on this subject based on the ideXlab platform.
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Tyrosine nitration of Prostacyclin Synthase is associated with enhanced retinal cell apoptosis in diabetes
The American Journal of Pathology, 2011Co-Authors: Ming-hui Zou, Mingkai Lin, Timothy J. Lyons, Zhonglin XieAbstract:The risk of diabetic retinopathy is associated with the presence of both oxidative stress and toxic eicosanoids. Whether oxidative stress actually causes diabetic retinopathy via the generation of toxic eicosanoids, however, remains unknown. The aim of the present study was to determine whether tyrosine nitration of Prostacyclin Synthase (PGIS) contributes to retinal cell death in vitro and in vivo. Exposure of human retinal pericytes to heavily oxidized and glycated LDL (HOG-LDL), but not native forms of LDL (N-LDL), for 24 hours significantly increased pericyte apoptosis, accompanied by increased tyrosine nitration of PGIS and decreased PGIS activity. Inhibition of the thromboxane receptor or cyclooxygenase-2 dramatically attenuated HOG-LDL–induced apoptosis without restoring PGIS activity. Administration of superoxide dismutase (to scavenge superoxide anions) or l-NG-nitroarginine methyl ester (l-NAME, a nonselective nitric oxide Synthase inhibitor) restored PGIS activity and attenuated pericyte apoptosis. In Akita mouse retinas, diabetes increased intraretinal levels of oxidized LDL and glycated LDL, induced PGIS nitration, enhanced apoptotic cell death, and impaired blood–retinal barrier function. Chronic administration of tempol, a superoxide scavenger, reduced intraretinal oxidized LDL and glycated LDL levels, PGIS nitration, and retina cell apoptosis, thereby preserving the integrity of blood–retinal barriers. In conclusion, oxidized LDL-mediated PGIS nitration and associated thromboxane receptor stimulation might be important in the initiation and progression of diabetic retinopathy.
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Peroxynitrite and protein tyrosine nitration of Prostacyclin Synthase.
Prostaglandins & Other Lipid Mediators, 2006Co-Authors: Ming-hui ZouAbstract:Abstract When working on the regulation of Prostacyclin Synthase (PGIS), we found that PGIS was selectively inhibited by peroxynitrite (ONOO − ), a potent oxidant formed by the combination of superoxide anion and nitric oxide (NO) at a rate of diffusion-controlled. None of the cellular antioxidants studied (i.e. GSH, Vitamins C and E, and others) prevented the inhibition of ONOO − on PGIS. This unexpected behavior was explained by a catalytic reaction of the iron-thiolate center of PGIS with ONOO − anion. In contrast, ONOO − activated both thromboxane A 2 -Synthase and cyclooxygenases. In addition, we demonstrated that sub-micromolar levels of ONOO − inhibited PGI 2 -dependent vasorelaxation and triggered a PGH 2 -dependent vasospasm, indicating that ONOO − increased PGH 2 formation as a consequence of PGIS nitration. We have subsequently demonstrated that endogenous ONOO − caused PGIS nitration and TxA 2 activation in several diseased conditions such as atherosclerotic vessels, hypoxia-reperfusion injury, cytokines-treated cells, diabetes, as well as hypertension. Since NO is produced physiologically it seems that excessive formation of superoxide not only eliminates the vasodilatory, growth-inhibiting, anti-thrombotic and anti-adhesive effects of NO and PGI 2 but also allows and promotes an action of the potent vasoconstrictor, prothrombotic agent, growth promoter, and leukocyte adherer, PGH 2 . We conclude that the nitration of PGIS nitration might be a new pathogenic mechanism for superoxide-induced endothelium dysfunction often observed in vascular diseases such as atherosclerosis, hypertension, ischemia, endotoxic shock, and diabetes.
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insulin resistance reduces arterial Prostacyclin Synthase and enos activities by increasing endothelial fatty acid oxidation
Journal of Clinical Investigation, 2006Co-Authors: Diane Edelstein, Ming-hui Zou, Silvana Obici, Ninon Higham, Michael BrownleeAbstract:Insulin resistance markedly increases cardiovascular disease risk in people with normal glucose tolerance, even after adjustment for known risk factors such as LDL, triglycerides, HDL, and systolic blood pressure. In this report, we show that increased oxidation of FFAs in aortic endothelial cells without added insulin causes increased production of superoxide by the mitochondrial electron transport chain. FFA-induced overproduction of superoxide activated a variety of proinflammatory signals previously implicated in hyperglycemia-induced vascular damage and inactivated 2 important antiatherogenic enzymes, Prostacyclin Synthase and eNOS. In 2 nondiabetic rodent models - insulin-resistant, obese Zucker (fa/fa) rats and high-fat diet-induced insulin-resistant mice - inactivation of Prostacyclin Synthase and eNOS was prevented by inhibition of FFA release from adipose tissue; by inhibition of the rate-limiting enzyme for fatty acid oxidation in mitochondria, carnitine palmitoyltransferase I; and by reduction of superoxide levels. These studies identify what we believe to be a novel mechanism contributing to the accelerated atherogenesis and increased cardiovascular disease risk occurring in people with insulin resistance.
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cd40 ligand dependent tyrosine nitration of Prostacyclin Synthase in vivo
Circulation, 2005Co-Authors: Bradley J. Davis, Ming-hui ZouAbstract:Background— Cells in human atherosclerotic lesions express the immune mediator CD40 and its ligand, CD40L, but the mechanisms and the mediators by which CD40L contributes to atherosclerosis are poorly defined. Here, we show how CD40L increases vascular inflammation and thrombosis via tyrosine nitration and inhibition of Prostacyclin Synthase (PGIS), an enzyme with antithrombotic, antiproliferative, and dilatory functions in the normal vasculature. Methods and Results— Exposure of cultured human aortic endothelial cells to clinically relevant concentrations of CD40L (20 to 80 ng/mL) dose-dependently increased the production of superoxide (O2·−), decreased nitric oxide (NO) bioactivity, and increased PGIS nitration. Furthermore, inhibition of CD40 expression by small interfering RNA blocked the effects of CD40L on O2·−, NO bioactivity, and PGIS nitration, which indicates a specific effect of CD40L. In addition, either depletion of mitochondria (ρ0 cells, ie, mitochondria-depleted cells, to prevent mitochond...
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CD40 Ligand–Dependent Tyrosine Nitration of Prostacyclin Synthase In Vivo
Circulation, 2005Co-Authors: Bradley J. Davis, Ming-hui ZouAbstract:Background— Cells in human atherosclerotic lesions express the immune mediator CD40 and its ligand, CD40L, but the mechanisms and the mediators by which CD40L contributes to atherosclerosis are poorly defined. Here, we show how CD40L increases vascular inflammation and thrombosis via tyrosine nitration and inhibition of Prostacyclin Synthase (PGIS), an enzyme with antithrombotic, antiproliferative, and dilatory functions in the normal vasculature. Methods and Results— Exposure of cultured human aortic endothelial cells to clinically relevant concentrations of CD40L (20 to 80 ng/mL) dose-dependently increased the production of superoxide (O2·−), decreased nitric oxide (NO) bioactivity, and increased PGIS nitration. Furthermore, inhibition of CD40 expression by small interfering RNA blocked the effects of CD40L on O2·−, NO bioactivity, and PGIS nitration, which indicates a specific effect of CD40L. In addition, either depletion of mitochondria (ρ0 cells, ie, mitochondria-depleted cells, to prevent mitochond...
Volker Ullrich - One of the best experts on this subject based on the ideXlab platform.
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Purification and characterization of recombinant human Prostacyclin Synthase.
Journal of biochemistry, 2004Co-Authors: Masayuki Wada, Chieko Yokoyama, Volker Ullrich, Toshihisa Hatae, Manabu Shimonishi, Masahiko Nakamura, Yoshio Imai, Tadashi TanabeAbstract:Prostacyclin Synthase (PGIS), which catalyzes the conversion of prostaglandin (PG) H(2) to Prostacyclin (PGI(2)), is a member of the cytochrome P-450 (P450) superfamily, CYP8A1. To study the enzymatic and protein characteristics of human PGIS, the enzyme was overexpressed in Spodoptera frugiperda 21 (Sf21) cells using the baculovirus expression system. PGIS was expressed in the microsomes of the infected Sf21 cells after culture in 5 microg/ml hematin-supplemented medium for 72 h. The holoenzyme was isolated from the solubilized microsomal fraction by calcium phosphate gel absorption and purified to homogeneity by DEAE-Sepharose and hydroxyapatite column chromatography. The K(m) and V(max) values of the purified human PGIS for PGH(2) were 30 microM and 15 micromol/min/mg of protein at 24 degrees C, respectively. The optical absorption and EPR spectra of the enzyme revealed the characteristics of a low-spin form of P450 in the oxidized state. The carbon monoxide-reduced difference spectrum, however, exhibited a peak at 418 nm rather than 450 nm. The addition of a PGH(2) analogue, U46619, to the enzyme produced an oxygen-ligand type of the difference spectrum with maximum absorption at 407 nm and minimum absorption at 430 nm. Treatment with another PGH(2) analogue, U44069, produced a peak at 387 nm and a trough at 432 nm in the spectrum (Type I), while treatment with tranylcypromine, a PGIS inhibitor, produced a peak at 434 nm and a trough at 412 nm (Type II). A Cys441His mutant of the enzyme possessed no heme-binding ability or enzyme activity. Thus, we succeeded in obtaining a sufficient amount of the purified recombinant human PGIS from infected insect cells for spectral analyses that has high specific activity and the characteristics of a P450, indicating substrate specificity.
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Nitration of Prostacyclin Synthase: mechanism and physiological implications
International Congress Series, 2002Co-Authors: Volker Ullrich, Andreas Daiber, Markus Bachschmid, Ming-hui ZouAbstract:Abstract In a series of investigations, we have provided evidence that Prostacyclin Synthase can be nitrated and inactivated under physiological conditions. Here, we address some pertinent questions on this process like the cause for selectivity, the cellular machinery of nitration, ways of its inhibition and a possible physiological or pathophysiological significance. The underlying mechanism of heme-thiolate catalysis explains the selectivity of peroxynitrite for Prostacyclin (PGI2)1 Synthase. The cellular conditions primarily require superoxide formation by a tightly controlled release, distinct from autoxidation processes. None of the well-known inhibitors of peroxynitrite-mediated nitrations and hydroxylations could block PGI2 Synthase nitration except ebselen, which, however, was ineffective in the presence of SH-groups. A physiological role may lie in the ability of superoxide to eliminate NO and, subsequently, PGI2 as mediators of vessel relaxation, of anti-adhesion properties and of differentiation. At the same time, the remaining Prostacyclin endoperoxide (PGH2) causes constriction, adhesion and proliferation through activation of the thromboxane A2/PGH2 receptor. The smooth muscle-dependent formation of PGE2 from PGH2 will also stimulate P-selectin release for the immune response. Pathophysiological conditions mediated by peroxynitrite (PN) may prevail after activation of early immediate genes and formation of peroxynitrite at much higher levels than required for endothelial dysfunction.
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PEROXYNITRITE INACTIVATES Prostacyclin Synthase BY HEME–THIOLATE-CATALYZED TYROSINE NITRATION
Drug metabolism reviews, 1999Co-Authors: Ming-hui Zou, Akin Yesilkaya, Volker UllrichAbstract:Previous work has shown a sensitive inhibition of Prostacyclin Synthase activity by peroxynitrite as well as by superoxide in the presence of NO donors. Neither superoxide nor NO alone nor decomposed peroxynitrite is effective. The inhibition of activity was paralleled by a nitration of a tyrosine residue and both could be prevented by a stable substrate analog. The same IC50 value for peroxynitrite was also found for the cellular Prostacyclin activity in endothelial and kidney mesangial cells, indicating that the antioxidant potential of the cell cannot prevent the inactivation. Aortic tissue shows a co-localization of Prostacyclin Synthase and nitrotyrosine staining after treatment of the tissue with 1 μM peroxynitrite. It can be speculated that this pathway of enzyme nitration is of pathophysiological significance.
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peroxynitrite inactivates Prostacyclin Synthase by heme thiolate catalyzed tyrosine nitration
Drug Metabolism Reviews, 1999Co-Authors: Ming-hui Zou, Akin Yesilkaya, Volker UllrichAbstract:Previous work has shown a sensitive inhibition of Prostacyclin Synthase activity by peroxynitrite as well as by superoxide in the presence of NO donors. Neither superoxide nor NO alone nor decomposed peroxynitrite is effective. The inhibition of activity was paralleled by a nitration of a tyrosine residue and both could be prevented by a stable substrate analog. The same IC50 value for peroxynitrite was also found for the cellular Prostacyclin activity in endothelial and kidney mesangial cells, indicating that the antioxidant potential of the cell cannot prevent the inactivation. Aortic tissue shows a co-localization of Prostacyclin Synthase and nitrotyrosine staining after treatment of the tissue with 1 μM peroxynitrite. It can be speculated that this pathway of enzyme nitration is of pathophysiological significance.
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Selective nitration of Prostacyclin Synthase and defective vasorelaxation in atherosclerotic bovine coronary arteries.
The American journal of pathology, 1999Co-Authors: Ming-hui Zou, Marcel Leist, Volker UllrichAbstract:Prostacyclin Synthase (PCS) is an enzyme with antithrombotic, antiproliferative, and dilatory functions in the normal vasculature, and inactivation of PCS by tyrosine nitration may favor atherosclerotic processes. Here, we show that PCS is nitrated and inactivated in early stage atherosclerotic lesions (focal intimal thickenings). Immunoprecipitation with antibodies raised against nitrotyrosine yielded PCS as the main nitrated protein in blood vessels. Moreover, we identified two nitrated degradation products of PCS with molecular mass of 30 and 46 kd, which were selective for atherosclerotic tissue. Agonist (acetylcholine, angiotensin II)-induced Prostacyclin formation was decreased in atherosclerotic vessels compared with normal tissue, whereas PGE 2 formation was increased and cyclooxygenase activity remained unchanged. A selective loss of PCS activity was confirmed by direct measurement of enzymatic activity. In line with this, we observed defective relaxation of early atherosclerotic vessels following vasoconstrictive stimulation. This functional impairment was completely reversed by coincubation with an antagonist of the thromboxane/PGH 2 receptor but not by a thromboxane Synthase inhibitor. These data suggest that reduced PCS activity in atherosclerotic arteries prevents the rapid use of PGH 2 , which accumulates and acts as an agonist on the vasoconstrictive thromboxane receptor.
Tomohiro Nakayama - One of the best experts on this subject based on the ideXlab platform.
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Prostacyclin Synthase gene: genetic polymorphisms and prevention of some cardiovascular diseases.
Current medicinal chemistry. Cardiovascular and hematological agents, 2005Co-Authors: Tomohiro NakayamaAbstract:Prostacyclin (PGI2) inhibits platelet aggregation and vasoconstriction. Prostacyclin Synthase (PGIS), a catalyst of PGI2 synthesis from prostaglandin H2, is widely distributed and predominantly found in vascular endothelial and smooth muscle cells. The PGIS gene is localized to 20q13.11-13, and a candidate gene for cardiovascular disease. We discovered mutations and polymorphisms in this gene and reported that they were associated with essential hypertension, myocardial infarction and cerebral infarction. These results suggest that PGI2 function depends on the different alleles of the PGIS gene and that they may influence the risk of cardiovascular diseases. Thus, individualized management strategies, such as administration of PGI2 analog, could be selected for variants of this gene to help prevent the development of cardiovascular diseases.
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Haplotype analysis of the Prostacyclin Synthase gene and essential hypertension.
Hypertension research : official journal of the Japanese Society of Hypertension, 2003Co-Authors: Tomohiro Nakayama, Masayoshi Soma, Katsuo Kanmatsuse, Akira Haketa, Noriko Aoi, Kotoko Kosuge, Mikano Sato, Shinichiro KokubunAbstract:Previously, we discovered 3 polymorphisms in the Prostacyclin Synthase (PGIS) gene: 1) T-192G, in the 5-flanking region, a novel single-nucleotide polymorphism (SNP) that is not associated with essential hypertension (EH); 2) a variable number of tandem repeat (VNTR) polymorphism, 6 nucleotides upstream from the ATG start codon, that is associated with risk of cerebral infarction; and 3) C1117A, in exon 8, an SNP that does not cause an amino acid change in codon 373, and that is associated with risk of myocardial infarction (MI). The purpose of the present study was to establish haplotypes of the PGIS gene consisting of these 3 polymorphisms, and to assess the association between these haplotypes and EH. We detected 19 haplotypes. There was no significant difference in the overall distribution of haplotypes between EH and normotensive subjects. To summarize, we successfully identified haplotypes of the PGIS gene, and these haplotypes were not associated with EH.
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Splicing mutation of the Prostacyclin Synthase gene in a family associated with hypertension.
Biochemical and biophysical research communications, 2002Co-Authors: Tomohiro Nakayama, Masayoshi Soma, Katsuo Kanmatsuse, Noriko Aoi, Kotoko Kosuge, Mikano Sato, Shinichiro Kokubun, Yoshiyasu Watanabe, Buaijiaer Hasimu, Jason D MarrowAbstract:Prostacyclin inhibits platelet aggregation, smooth muscle cell proliferation, and vasoconstriction. The Prostacyclin Synthase (PGIS) gene is a candidate gene for cardiovascular disease. The purpose of this study was to locate possible mutations in the PGIS gene related to hypertension and cerebral infarction. Using the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) method, we discovered a T to C transition at the +2 position of the splicing donor site of intron 9 in patients with essential hypertension (EH). In vitro expression analysis of an allelic minigene consisting of exons 8-10 revealed that the nucleotide transition causes skipping of exon 9. This in turn alters the translational reading frame of exon 10 and introduces a premature stop codon (TGA). A three-dimensional model shows that the splice site mutation produces a truncated protein with a deletion in the heme-binding region. This splice site mutation was found in only one subject in 200 EH patients and 200 healthy controls. Analysis of the patient's family members revealed the mutation in two of the three siblings. The urinary excretion of Prostacyclin metabolites in subjects with the mutation was significantly decreased. All subjects displaying the splice site mutation in the PGIS gene were hypertensive. In this study, we report a novel splicing mutation in the PGIS gene, which is associated with hypertension in a family. It is thought that this mechanism may involve in the pathophysiology of their hypertension.
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Association Study between a Novel Single Nucleotide Polymorphism of the Promoter Region of the Prostacyclin Synthase Gene and Essential Hypertension
Hypertension research : official journal of the Japanese Society of Hypertension, 2002Co-Authors: Tomohiro Nakayama, Dolkun Rahmutula, Masayoshi Soma, Hideko Tobe, Jiro Uwabo, Masako Kunimoto, Noriko Aoi, Kotoko Kosuge, Mikano Sato, Katsuo KanmatsuseAbstract:The purpose of this study was to investigate whether an association exists between the promoter region of the Prostacyclin Synthase gene and essential hypertension (EH). Using the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) method, we discovered a novel single nucleotide polymorphism (SNP), T-192G, in the 5′-flanking region. We performed an association study using the SNP in 200 patients and 200 controls. The allele frequency distribution in the two groups was not significantly different. Thus, this SNP in the PGIS gene is not associated with EH. (Hypertens Res 2002; 25: 65-68)
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Association of a novel single nucleotide polymorphism of the Prostacyclin Synthase gene with myocardial infarction
American heart journal, 2002Co-Authors: Tomohiro Nakayama, Dolkun Rahmutula, Masayoshi Soma, Jiro Uwabo, Mikano Sato, Satoshi Saito, Junko Honye, Junji Yajima, Yukie Kaneko, Noriko AoiAbstract:Abstract Background Myocardial infarction (MI) is a complex multifactorial and polygenic disorder that is thought to result from an interaction between an individual's genetic makeup and various environmental factors. The purpose of this study was to investigate the association between a novel single nucleotide polymorphism in the Prostacyclin Synthase gene and MI. Methods and Results By the use of polymerase chain reaction-single-strand conformation polymorphism analysis, we identified a single nucleotide polymorphism, C1117A, in exon 8. This nucleotide change did not cause an amino acid change in codon 373. We performed an association study of the polymorphism in 138 patients and 130 healthy control subjects. Multiple logistic linear regression analysis showed the genotype distributions were significantly different between the control group and the MI group (odds ratio, 2.12; 95% CI, 1.47-3.05, P =.04). The C/C genotype was found more frequently in the MI group than in the control group. Conclusions We conclude that the C1117A polymorphism in exon 8 is associated with risk for MI and may be a genetic marker of MI in Japanese persons. (Am Heart J 2002;143:797-801.)
Tadashi Tanabe - One of the best experts on this subject based on the ideXlab platform.
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Acceleration of wound healing by combined gene transfer of hepatocyte growth factor and Prostacyclin Synthase with Shima Jet.
Gene therapy, 2006Co-Authors: Yasuo Kunugiza, Tadashi Tanabe, Naruya Tomita, Yoshiaki Taniyama, Tetsuya Tomita, Mariana Kiomy Osako, Katsuto Tamai, Y Kaneda, Hideki Yoshikawa, Ryuichi MorishitaAbstract:Although skin diseases are one of the target diseases for gene therapy, there has been no practical gene transfer method. First, we examined gene transfer efficiency of the spring-powered jet injector, Shima Jet, which was originally developed as a non-needle jet injector of insulin. Local gene expression was about 100 times higher when the luciferase plasmid was transferred by the Shima Jet than by a needle. Gene transfer of β-galactosidase revealed gene expression in the epidermis. Based on these results, we then examined the potential of gene therapy using the Shima Jet for wound healing. An increase of cellular proliferation of the epidermis and the number of microvessels in the granulation tissue was observed after hepatocyte growth factor (HGF) gene transfer. An increase in blood flow around the wound was observed after Prostacyclin Synthase (PGIS) gene transfer. Moreover, promotion on wound healing was observed in HGF gene transferred group, and further promotion was observed in combined gene transferred group as assessed by measuring wound area. These results indicate that co-transfer of HGF and PGIS genes by the Shima Jet could be an effective strategy to wound healing.
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Purification and characterization of recombinant human Prostacyclin Synthase.
Journal of biochemistry, 2004Co-Authors: Masayuki Wada, Chieko Yokoyama, Volker Ullrich, Toshihisa Hatae, Manabu Shimonishi, Masahiko Nakamura, Yoshio Imai, Tadashi TanabeAbstract:Prostacyclin Synthase (PGIS), which catalyzes the conversion of prostaglandin (PG) H(2) to Prostacyclin (PGI(2)), is a member of the cytochrome P-450 (P450) superfamily, CYP8A1. To study the enzymatic and protein characteristics of human PGIS, the enzyme was overexpressed in Spodoptera frugiperda 21 (Sf21) cells using the baculovirus expression system. PGIS was expressed in the microsomes of the infected Sf21 cells after culture in 5 microg/ml hematin-supplemented medium for 72 h. The holoenzyme was isolated from the solubilized microsomal fraction by calcium phosphate gel absorption and purified to homogeneity by DEAE-Sepharose and hydroxyapatite column chromatography. The K(m) and V(max) values of the purified human PGIS for PGH(2) were 30 microM and 15 micromol/min/mg of protein at 24 degrees C, respectively. The optical absorption and EPR spectra of the enzyme revealed the characteristics of a low-spin form of P450 in the oxidized state. The carbon monoxide-reduced difference spectrum, however, exhibited a peak at 418 nm rather than 450 nm. The addition of a PGH(2) analogue, U46619, to the enzyme produced an oxygen-ligand type of the difference spectrum with maximum absorption at 407 nm and minimum absorption at 430 nm. Treatment with another PGH(2) analogue, U44069, produced a peak at 387 nm and a trough at 432 nm in the spectrum (Type I), while treatment with tranylcypromine, a PGIS inhibitor, produced a peak at 434 nm and a trough at 412 nm (Type II). A Cys441His mutant of the enzyme possessed no heme-binding ability or enzyme activity. Thus, we succeeded in obtaining a sufficient amount of the purified recombinant human PGIS from infected insect cells for spectral analyses that has high specific activity and the characteristics of a P450, indicating substrate specificity.
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Enhanced Therapeutic Angiogenesis by Cotransfection of Prostacyclin Synthase Gene or Optimization of Intramuscular Injection of Naked Plasmid DNA
Circulation, 2003Co-Authors: Kazuya Hiraoka, Chieko Yokoyama, Tadashi Tanabe, Toshio Ogihara, Hiromi Koike, Yasufumi Kaneda, Seiji Yamamoto, Naruya Tomita, Takashi Aikou, Ryuichi MorishitaAbstract:Background— Although clinical trials of therapeutic angiogenesis by angiogenic growth factors with intramuscular injection of naked plasmid DNA have been successful, there are still unresolved problems such as low transfection efficiency. From this viewpoint, we performed the following modifications: (1) combination with vasodilation using Prostacyclin and (2) changing the agents or volume of naked plasmid DNA in vivo. Methods and Results— First, we examined cotransfection of the VEGF gene with the Prostacyclin Synthase gene in a mouse hindlimb ischemia model. Cotransfection of the VEGF gene with the Prostacyclin Synthase gene resulted in a further increase in blood flow and capillary density compared with single VEGF gene. Similar results were obtained with other angiogenic growth factors, such as hepatocyte growth factor (HGF). Alternatively, we changed the injection volume of the solution of plasmid DNA. Luciferase activity was increased in a volume-dependent manner. An increase in injection volume at ...
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Enhanced angiogenesis and improvement of neuropathy by cotransfection of human hepatocyte growth factor and Prostacyclin Synthase gene
FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 2003Co-Authors: Hiromi Koike, Chieko Yokoyama, Tadashi Tanabe, Toshio Ogihara, Ryuichi Morishita, Sohta Iguchi, Motokuni Aoki, Kunio Matsumoto, Toshikazu Nakamura, Yasufumi KanedaAbstract:The current therapeutic angiogenesis strategy to treat ischemic disease by using angiogenic growth factors has been limited to use of a single gene. However, as vasodilator substances such as Prostacyclin are widely used for the treatment of peripheral arterial disease, it might be useful to combine angiogenesis with vasodilation of new vessels. In a mouse hind limb ischemia model, cotransfection of the hepatocyte growth factor (HGF) gene with the Prostacyclin Synthase gene demonstrated a further increase in blood flow and capillary density compared with a single gene. Even in the rabbit ischemia model, cotransfection of HGF plasmid with the Prostacyclin Synthase gene demonstrated a further increase in angiogenic activity compared with HGF alone. Because peripheral neuropathy due to diabetes is common for significant morbidity, we examined the hypothesis that experimental diabetic neuropathy can be reversed by HGF and Prostacyclin Synthase genes. Severe peripheral neuropathy, characterized by significant slowing of nerve conduction velocity compared with nondiabetic control animals, was ameliorated. Overall, cotransfection of the Prostacyclin Synthase and HGF genes is more effective than single-gene transfection to stimulate angiogenesis, and it significantly improved neuropathy. These data provide important information relating to the clinical application of therapeutic angiogenesis to treat peripheral arterial disease.
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cyclic mechanical stretch augments Prostacyclin production in cultured human uterine myometrial cells from pregnant women possible involvement of up regulation of Prostacyclin Synthase expression
The Journal of Clinical Endocrinology and Metabolism, 2002Co-Authors: Daizo Korita, Chieko Yokoyama, Tadashi Tanabe, Norimasa Sagawa, Hiroaki Itoh, Shigeo Yura, Masahiro Yoshida, Kazuyo Kakui, Maki Takemura, Shingo FujiiAbstract:Prostacyclin (PGI2), a potent smooth muscle relaxant, is a major prostaglandin secreted from human myometrium. The concentrations of PGI2 metabolites in the maternal plasma were reported to be elevated during pregnancy, especially in labor. To clarify the mechanism in PGI2 secretion from the myometrium, we first investigated the protein expression of cytosolic phospholipase A2, cyclooxygenase (COX)-1, COX-2, and Prostacyclin Synthase (PGIS) in the human uterine myometrium at various gestational ages before labor. To elucidate the involvement of labor in the increase in PGI2 production during labor, we next examined the effect of labor-like cyclic mechanical stretch on PGI2 production by cultured human myometrial cells. Pregnancy specifically increased COX-1 and PGIS protein expression in the myometrial tissues before labor (P < 0.01 for both). Cyclic mechanical stretch augmented PGIS promoter activity, via activation of activator protein-1 site, and PGIS mRNA and protein expression in cultured human myome...
Katsuo Kanmatsuse - One of the best experts on this subject based on the ideXlab platform.
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Haplotype analysis of the Prostacyclin Synthase gene and essential hypertension.
Hypertension research : official journal of the Japanese Society of Hypertension, 2003Co-Authors: Tomohiro Nakayama, Masayoshi Soma, Katsuo Kanmatsuse, Akira Haketa, Noriko Aoi, Kotoko Kosuge, Mikano Sato, Shinichiro KokubunAbstract:Previously, we discovered 3 polymorphisms in the Prostacyclin Synthase (PGIS) gene: 1) T-192G, in the 5-flanking region, a novel single-nucleotide polymorphism (SNP) that is not associated with essential hypertension (EH); 2) a variable number of tandem repeat (VNTR) polymorphism, 6 nucleotides upstream from the ATG start codon, that is associated with risk of cerebral infarction; and 3) C1117A, in exon 8, an SNP that does not cause an amino acid change in codon 373, and that is associated with risk of myocardial infarction (MI). The purpose of the present study was to establish haplotypes of the PGIS gene consisting of these 3 polymorphisms, and to assess the association between these haplotypes and EH. We detected 19 haplotypes. There was no significant difference in the overall distribution of haplotypes between EH and normotensive subjects. To summarize, we successfully identified haplotypes of the PGIS gene, and these haplotypes were not associated with EH.
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Splicing mutation of the Prostacyclin Synthase gene in a family associated with hypertension.
Biochemical and biophysical research communications, 2002Co-Authors: Tomohiro Nakayama, Masayoshi Soma, Katsuo Kanmatsuse, Noriko Aoi, Kotoko Kosuge, Mikano Sato, Shinichiro Kokubun, Yoshiyasu Watanabe, Buaijiaer Hasimu, Jason D MarrowAbstract:Prostacyclin inhibits platelet aggregation, smooth muscle cell proliferation, and vasoconstriction. The Prostacyclin Synthase (PGIS) gene is a candidate gene for cardiovascular disease. The purpose of this study was to locate possible mutations in the PGIS gene related to hypertension and cerebral infarction. Using the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) method, we discovered a T to C transition at the +2 position of the splicing donor site of intron 9 in patients with essential hypertension (EH). In vitro expression analysis of an allelic minigene consisting of exons 8-10 revealed that the nucleotide transition causes skipping of exon 9. This in turn alters the translational reading frame of exon 10 and introduces a premature stop codon (TGA). A three-dimensional model shows that the splice site mutation produces a truncated protein with a deletion in the heme-binding region. This splice site mutation was found in only one subject in 200 EH patients and 200 healthy controls. Analysis of the patient's family members revealed the mutation in two of the three siblings. The urinary excretion of Prostacyclin metabolites in subjects with the mutation was significantly decreased. All subjects displaying the splice site mutation in the PGIS gene were hypertensive. In this study, we report a novel splicing mutation in the PGIS gene, which is associated with hypertension in a family. It is thought that this mechanism may involve in the pathophysiology of their hypertension.
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Association Study between a Novel Single Nucleotide Polymorphism of the Promoter Region of the Prostacyclin Synthase Gene and Essential Hypertension
Hypertension research : official journal of the Japanese Society of Hypertension, 2002Co-Authors: Tomohiro Nakayama, Dolkun Rahmutula, Masayoshi Soma, Hideko Tobe, Jiro Uwabo, Masako Kunimoto, Noriko Aoi, Kotoko Kosuge, Mikano Sato, Katsuo KanmatsuseAbstract:The purpose of this study was to investigate whether an association exists between the promoter region of the Prostacyclin Synthase gene and essential hypertension (EH). Using the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) method, we discovered a novel single nucleotide polymorphism (SNP), T-192G, in the 5′-flanking region. We performed an association study using the SNP in 200 patients and 200 controls. The allele frequency distribution in the two groups was not significantly different. Thus, this SNP in the PGIS gene is not associated with EH. (Hypertens Res 2002; 25: 65-68)
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Polymorphism of the promoter region of Prostacyclin Synthase gene is not related to essential hypertension.
American Journal of Hypertension, 2001Co-Authors: Tomohiro Nakayama, Yoichi Izumi, Masayoshi Soma, Duolikun Rehemudula, Yukie Takahashi, Hideko Tobe, Jiro Uwabo, Masako Kunimoto, Mikano Sato, Katsuo KanmatsuseAbstract:An impaired synthesis of Prostacyclin has been implicated in the development of essential hypertension (EH). We therefore investigated whether there is an association between the Prostacyclin Synthase (PGIS) gene and EH using a variable number of tandem repeats (VNTR) polymorphism in the promoter region that influences transcriptional activity of this gene. A total of 125 patients with EH and 125 age-matched subjects with normal blood pressure were studied. The number of VNTR of the five alleles ranged from 3 to 7 repeats in the 250 unrelated Japanese subjects. The allele frequency distribution in the two groups were not significantly different. Thus, this VNTR polymorphism in the PGIS gene is not associated with EH.
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Association of 5' upstream promoter region of Prostacyclin Synthase gene variant with cerebral infarction.
American journal of hypertension, 2000Co-Authors: Tomohiro Nakayama, Masayoshi Soma, Duolikun Rehemudula, Yukie Takahashi, Hideko Tobe, Mikano Satoh, Jiro Uwabo, Masako Kunimoto, Katsuo KanmatsuseAbstract:The aim of this study was to investigate whether there is an association between the promoter region of the Prostacyclin Synthase gene and cerebral infarction (CI). Using the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) method, we found a variable-number tandem repeat polymorphism in the 5'-upstream promoter region of the Prostacyclin Synthase gene. This region contains transcriptional factors-binding sites of Spl (CCCGCC) and AP-2 (CCGCCAGCCCC). The alleles varied in size from three to seven repeats of nine base pairs (bp). We performed an association study using the polymorphism in 111 patients and 152 control subjects. The transcriptional activity of the abnormal promoter region allele was determined by luciferase assay. The overall distribution of alleles differed significantly between both groups. Logistic linear regression analysis revealed the small number repeat allele to be found more frequently with CI. Transcriptional activity increased with increasing numbers of repeats. This study provides consistent support for the association between CI and the PGIS gene.
