Biomedical subjects

Jun Yoshioka

Publications and source records attributed to Jun Yoshioka.

12 recordsLinked to original sources

PubMed2006

Nitric oxide-dependent suppression of thioredoxin-interacting protein expression enhances thioredoxin activity.

OBJECTIVE: Cellular redox balance is regulated by enzymatic and nonenzymatic systems and freely diffusible nitric oxide (NO) promotes antioxidative mechanisms. We show the NO-dependent transcriptional regulation of the antioxidative thioredoxin system. METHODS AND RESULTS: Incubation of rat pulmonary artery smooth muscle cells (RPaSMC) with the NO donor compound S-nitroso-glutathione (GSNO, 100 micromol/L) suppressed thioredoxin-interacting protein (Txnip), an inhibitor of thioredoxin function, by 71+/-18% and enhanced thioredoxin reductase 2.7+/-0.2 fold (n=6; both P<0.001 versus control). GSNO increased thioredoxin activity (1.9+/-0.5-fold after 4 hours; P<0.05 versus control). Promoter deletion analysis revealed that NO suppression of Txnip transcription is mediated by cis-regulatory elements between -1777 and -1127 bp upstream of the start codon. Hyperglycemia induced Txnip promoter activity (3.9+/-0.2-fold; P<0.001) and abolished NO effects (-37.4+/-1.0% at 5.6 mmol/L glucose versus 12.4+/-2.1% at 22.4 mmol/L glucose; P<0.05). Immunoprecipitation experiments demonstrated that GSNO stimulation and mutation of thioredoxin at Cys69, a site of nitrosylation, had no effect on the Txnip/thioredoxin interaction. CONCLUSIONS: NO can regulate cellular redox state by changing expression of Txnip and thioredoxin reductase. This represents a novel antioxidative mechanism of NO independent of posttranslational protein S-nitrosylation of thioredoxin.

Animals↗

PubMed2006

The interaction of thioredoxin with Txnip. Evidence for formation of a mixed disulfide by disulfide exchange.

The thioredoxin system plays an important role in maintaining a reducing environment in the cell. Recently, several thioredoxin binding partners have been identified and proposed to mediate aspects of redox signaling, but the significance of these interactions is unclear in part due to incomplete understanding of the mechanism for thioredoxin binding. Thioredoxin-interacting protein (Txnip) is critical for regulation of glucose metabolism, the only currently known function of which is to bind and inhibit thioredoxin. We explored the mechanism of the Txnip-thioredoxin interaction and present evidence that Txnip and thioredoxin form a stable disulfide-linked complex. We identified two Txnip cysteines that are important for thioredoxin binding and showed that this interaction is consistent with a disulfide exchange reaction between oxidized Txnip and reduced thioredoxin. These cysteines are not conserved in the broader family of arrestin domain-containing proteins, and we demonstrate that the thioredoxin-binding property of Txnip is unique. These data suggest that Txnip is a target of reduced thioredoxin and provide insight into the potential role of Txnip as a redox-sensitive signaling protein.

3T3-L1 Cells↗

PubMed2005

Cardiomyocyte hypertrophy and degradation of connexin43 through spatially restricted autocrine/paracrine heparin-binding EGF.

Growth factor signaling can affect tissue remodeling through autocrine/paracrine mechanisms. Recent evidence indicates that EGF receptor transactivation by heparin-binding EGF (HB-EGF) contributes to hypertrophic signaling in cardiomyocytes. Here, we show that HB-EGF operates in a spatially restricted circuit in the extracellular space within the myocardium, revealing the critical nature of the local microenvironment in intercellular signaling. This highly localized microenvironment of HB-EGF signaling was demonstrated with 3D morphology, consistent with predictions from a computational model of EGF signaling. HB-EGF secretion by a given cardiomyocyte in mouse left ventricles led to cellular hypertrophy and reduced expression of connexin43 in the overexpressing cell and in immediately adjacent cells but not in cells farther away. Thus, HB-EGF acts as an autocrine and local paracrine cardiac growth factor that leads to loss of gap junction proteins within a spatially confined microenvironment. These findings demonstrate how cells can coordinate remodeling with their immediate neighboring cells with highly localized extracellular EGF signaling.

Analysis of Variance↗

PubMed2004

Hyperglycemia promotes oxidative stress through inhibition of thioredoxin function by thioredoxin-interacting protein.

Increased intracellular reactive oxygen species (ROS) contribute to vascular disease and pro-atherosclerotic effects of diabetes mellitus may be mediated by oxidative stress. Several ROS-scavenging systems tightly control cellular redox balance; however, their role in hyperglycemia-induced oxidative stress is unclear. A ubiquitous antioxidative mechanism for regulating cellular redox balance is thioredoxin, a highly conserved thiol reductase that interacts with an endogenous inhibitor, thioredoxin-interacting protein (Txnip). Here we show that hyperglycemia inhibits thioredoxin ROS-scavenging function through p38 MAPK-mediated induction of Txnip. Overexpression of Txnip increased oxidative stress, while Txnip gene silencing restored thioredoxin activity in hyperglycemia. Diabetic animals exhibited increased vascular expression of Txnip and reduced thioredoxin activity, which normalized with insulin treatment. These results provide evidence for the impairment of a major ROS-scavenging system in hyperglycemia. These studies implicate reduced thioredoxin activity through interaction with Txnip as an important mechanism for vascular oxidative stress in diabetes mellitus.

Animals↗

PubMed2004

Thioredoxin-interacting protein controls cardiac hypertrophy through regulation of thioredoxin activity.

BACKGROUND: Although cellular redox balance plays an important role in mechanically induced cardiac hypertrophy, the mechanisms of regulation are incompletely defined. Because thioredoxin is a major intracellular antioxidant and can also regulate redox-dependent transcription, we explored the role of thioredoxin activity in mechanically overloaded cardiomyocytes in vitro and in vivo. METHODS AND RESULTS: Overexpression of thioredoxin induced protein synthesis in cardiomyocytes (127+/-5% of controls, P<0.01). Overexpression of thioredoxin-interacting protein (Txnip), an endogenous thioredoxin inhibitor, reduced protein synthesis in response to mechanical strain (89+/-5% reduction, P<0.01), phenylephrine (80+/-3% reduction, P<0.01), or angiotensin II (80+/-4% reduction, P<0.01). In vivo, myocardial thioredoxin activity increased 3.5-fold compared with sham controls after transverse aortic constriction (P<0.01). Aortic constriction did not change thioredoxin expression but reduced Txnip expression by 40% (P<0.05). Gene transfer studies showed that cells that overexpress Txnip develop less hypertrophy after aortic constriction than control cells in the same animals (28.1+/-5.2% reduction versus noninfected cells, P<0.01). CONCLUSIONS: Thus, even though thioredoxin is an antioxidant, activation of thioredoxin participates in the development of pressure-overload cardiac hypertrophy, demonstrating the dual function of thioredoxin as both an antioxidant and a signaling protein. These results also support the emerging concept that the thioredoxin inhibitor Txnip is a critical regulator of biomechanical signaling.

Angiotensin II↗

PubMed2004

Oestrogen increases myocardial blood flow in men: assessment by 13N-ammonia positron emission tomography.

OBJECTIVES: Oestrogen has been shown to increase nitric oxide-mediated vasodilatation and modulate sympathetic tone in postmenopausal women. We investigated the acute effects of oestrogen on the coronary microcirculation in men. METHODS: Myocardial blood flow was quantified using N-ammonia positron emission tomography before and 20 min after an intravenous administration of conjugated oestrogen (10 mg) in nine healthy men. RESULTS: There were no significant changes in either systemic blood pressure or heart rate before and after oestrogen infusion. However, myocardial blood flow was increased by oestrogen from 0.88 +/-0.06 to 1.05+/-0.09 ml x g x min (P<0.05). Although serum levels of nitrite/nitrate (end products of nitric oxide) were not increased, circulating norepinephrine (noradrenaline) levels were decreased after the administration of oestrogen from 401+/-114 to 346+/-112 pg x ml (P<0.01). A significant negative correlation was found between myocardial blood flow and plasma norepinephrine levels (r=-0.67, P<0.05). CONCLUSIONS: A single administration of oestrogen enhanced myocardial circulation in healthy men. The oestrogen-induced increase in myocardial blood flow may be due to direct effects on the coronary circulation or through the modulation of norepinephrine levels, rather than the production of nitric oxide.

Adult↗

PubMed2003

Effect of a cleavage-resistant collagen mutation on left ventricular remodeling.

Matrix metalloproteinase-mediated degradation of type I collagen may play a role in cardiac remodeling after strain or injury. To explore this hypothesis, we used mice homozygous (r/r) for a targeted mutation in Col1a1; these mice synthesize collagen I that resists collagenase cleavage at Gly975-Leu976. A total of 64 r/r and 84 littermate wild-type mice (WT) underwent experimental pressure overload by transverse aortic constriction (TAC) or myocardial infarction (MI). Echocardiographic, hemodynamic, and histological parameters were evaluated up to 12 weeks after TAC or 21 days after MI. At 4 weeks after TAC, collagen levels, wall thickness, and echocardiographic parameters were similar in the 2 groups. At 12 weeks after TAC, r/r mice had smaller LV dimensions (ESD: 2.7+/-0.2 mm WT versus 1.7+/-0.2 mm r/r, P<0.013; EDD: 3.8+/-0.2 mm WT versus 3.1+/-0.1 mm r/r, P<0.013); better fractional shortening (30+/-2% WT versus 46+/-4% r/r; P<0.013); and lower LV/body weight ratios (7.3+/-0.6 WT and 5.1+/-0.5 r/r; P<0.013). Surprisingly, these differences were not accompanied by differences in collagen accumulation, myocyte cross-sectional areas, wall thickness, or microvessel densities. Furthermore, no differences in LV remodeling assessed by echocardiography, fibrosis, or hemodynamic parameters were found between r/r and WT mice after MI. Thus, a mutation that encodes a collagenase cleavage-resistant collagen I does not affect early LV remodeling after TAC or MI, suggesting that collagen cleavage at this site is not the mechanism by which metalloproteinases mediate LV remodeling. Collagen cleavage could, however, have a role in preservation of cardiac function in late remodeling by mechanisms independent of collagen accumulation. We were not able to detect collagen cleavage fragments, and could not, therefore, rule out the possibility of collagen cleavage at additional sites.

Animals↗

PubMed2002

Vitamin D3-upregulated protein-1 (VDUP-1) regulates redox-dependent vascular smooth muscle cell proliferation through interaction with thioredoxin.

Reactive oxygen species are important cellular signaling molecules, and thioredoxin (TRX) is a key regulator of cellular redox balance. We investigated the interaction of TRX with its endogenous inhibitor, vitamin D3-upregulated protein (VDUP)-1, in human aortic smooth muscle cells (SMCs). Adenoviral gene transfer of TRX enhanced TRX enzyme activity 2.7+/-0.4-fold (P<0.05 versus cells infected with adenoviral vector expressing green fluorescent protein [AdGFP]) and resulted in a 3.8+/-0.5-fold increase of cellular DNA synthesis as detected by methyl-[3H]thymidine incorporation (P<0.001). Platelet-derived growth factor (PDGF) also increased TRX enzyme activity 2.5+/-3.3-fold (P<0.05 versus no stimulation) and DNA synthesis 6.5+/-0.3-fold (P<0.001 versus no stimulation) without significant changes in TRX expression. PDGF and H2O2 time-dependently suppressed VDUP-1 expression (13-fold and 30-fold reduction after 1 hour, respectively; P<0.001), and this was inhibited by the cell-permeable antioxidants N-acetylcysteine and 4,5-dihydroxy-1,3-benzene-disulfonic acid (Tiron). Overexpression of VDUP-1 (AdVDUP-1) reduced TRX activity at baseline (-61+/-23% versus control cells, P<0.05) and abolished PDGF-induced TRX activity (-9+/-27% in AdVDUP-1-infected cells; P=NS versus control cells). In addition, overexpression of VDUP-1 blocked PDGF-induced DNA synthesis (1.3+/-0.4-fold increase in AdVDUP-1-infected cells versus 6.5+/-0.4-fold increase in AdGFP-infected cells, P<0.001). In conclusion, VDUP-1 has marked antiproliferative effects in SMCs through the suppression of TRX activity, suggesting that the regulation of VDUP-1 is a critical molecular switch in the transduction of pro-oxidant mitogenic signals. These data also demonstrate that activation of the reductase TRX plays a pivotal role in the redox-dependent proliferation of SMCs.

Active Transport, Cell Nucleus↗

PubMed2002

Myocardial viability assessment with gated SPECT Tc-99m tetrofosmin % wall thickening: comparison with F-18 FDG-PET.

OBJECT: This study was designed to assess the value of gated SPECT Tc-99m-tetrofosmin (TF) wall thickening (WT) in addition to TF exercise (Ex)/rest myocardial SPECT, in comparison with F-18 fluorodeoxyglucose (FDG)-PET. METHODS: The study population consisted of 33 patients with old myocardial infarction (27 men and 6 women; mean age, 62 +/- 8 years old). All patients underwent Ex/rest TF SPECT and glucose loading FDG-PET. Polar map images of Ex/rest TF were generated and divided into 24 segments for further analysis. We classified LV segments according to the exercise-rest perfusion scintigraphy. LV segments with less than 70% of the maximum TF activity on the exercise image were defined as stress-induced defects. Among these, the segments whose TF activity increased by 10% from exercise to rest images or exceeded 70% of the maximum uptake were defined as reversible (viable) defects. The remaining defects on the rest image were irreversible (non-viable) defect segments, and were considered for viability study on the basis of %WT. %WT was calculated according to the standard method: [(counts ES - counts ED)/counts ED] x 100. A viable segment on gated SPECT was defined as a segment whose %WT exceeded the lower limit of the normal value (mean - SD). PET viability was defined as FDG uptake exceeding 50% of the maximum count. RESULTS: Among the 792 segments evaluated in the 33 patients studied, there were 689 PET viable segments. Of the 689 segments analyzed, 198 (29%) were identified as having defects on Ex images. Among these defects, 55 (8%) were reversible or partially reversible, as evidenced by rest images, and 143 (21%) were irreversible. Of the irreversible segments on Ex/rest images, 106 (15%) demonstrated no apparent WT by gated TF SPECT, whereas 37 (6%) segments with irreversible defects did have apparent WT. Overall, the sensitivity of Ex/rest TF perfusion imaging was 79%. Sensitivity was improved from 79% to 85% by combining %WT and perfusion data, but specificity was reduced from 70% to 56%. CONCLUSION: %WT evaluated from gated TF imaging enhanced myocardial viability assessment in comparison with FDG-PET.

Aged↗

PubMedDate not supplied

Cardiovascular genomics.

The ongoing explosion of genomic information is revolutionizing cardiovascular science. More complete genomic maps allow easier identification of genes that cause monogenic inherited diseases. In addition, analyses of variations in gene expression in cardiovascular diseases are revealing new potential candidate genes as well as novel biomarkers for many common, multifactorial diseases. While experiments are revealing new pathophysiologic pathways, these genomic studies are also generating enormous amounts of data. Even as we learn how to sift through mountains of genomic information, new technologies in proteomics are rapidly advancing and promise to provide additional critical tools for diagnosis and for identifying new therapies.

Biomarkers↗

PubMedDate not supplied

Characteristics of regional myocardial stunning after exercise in gated myocardial SPECT.

BACKGROUND: A number of studies have demonstrated prolonged left ventricular (LV) global dysfunction after exercise-induced ischemia in gated myocardial single photon emission tomography (SPECT) as a manifestation of exercise-induced stunning. This study investigated the residual effects of exercise on postexercise LV regional function and its implications on the detection of stunning in gated SPECT. METHODS AND RESULTS: Fifty-three subjects with known or suspected coronary artery disease and 10 control subjects underwent myocardial SPECT according to a same-day exercise-rest protocol. Both postexercise and resting images were gated and acquired 1 hour after injection of technetium 99m tetrofosmin. The LV global ejection fraction and segmental systolic wall thickening were quantitated with the use of an automatic program. Segmental perfusion was assessed semiquantitatively on summed nongated tomograms. Wall thickening index (WTI), the ratio of systolic wall thickening of a segment to that of a corresponding control segment, was significantly lower after exercise than at rest in the reversible defect (RD) segments (0.66 +/- 0.24 vs 0.78 +/- 0.24; P <.0001). In patients with exercise-induced ischemia, the difference in WTI between rest and after exercise was significantly greater in the RD segments, which represented ischemia, than in the non-RD segments. Postexercise WTIs were not different from the resting values in subjects with no perfusion abnormalities or who had fixed defects (infarction). Significant postexercise dysfunction was present in 44% of the RD segments, compared with 5% of the normal and 3% of the fixed defect segments. Postexercise segmental dysfunction was correlated with the segmental reversibility score, the difference in defect scores between exercise and rest images (n = 82, Spearman rank correlation coefficient = -0.78, P <.0001). Among 19 patients with ischemia, 9 (47%) exhibited concurrent segmental and global dysfunction, but segmental dysfunction persisted in the absence of global dysfunction in 4 additional patients (21%). CONCLUSIONS: Significant postexercise LV regional dysfunction, consistent with the concept of stunning, occurs in the region of severe ischemia. The incidence and magnitude of regional stunning are determined by the severity of ischemia. For the detection of stunning in gated SPECT, LV regional dysfunction may be more sensitive than global dysfunction.

Adult↗

PubMedDate not supplied

Role of thioredoxin in cell growth through interactions with signaling molecules.

The thioredoxin system helps maintain a reducing environment in cells, but thioredoxin functions as more than simply an antioxidant. Thioredoxin functions depend on the protein's redox state, as determined by two conserved cysteines. Key biologic activities of thioredoxin include antioxidant, growth control, and antiapoptotic properties, resulting from interaction with target molecules including transcription factors. Mechanisms by which thioredoxin regulates cell growth include binding to signaling molecules such as apoptosis signal-regulating kinase-1 (ASK-1) and thioredoxin-interacting protein (Txnip). The molecular interplay between thioredoxin, ASK-1, and Txnip potentially influences cell growth and survival in diverse human diseases such as cancer, diabetes, and heart disease. In this review, we focus on the structure of thioredoxin and its functional regulation of cell growth through the interactions with signaling molecules.

Carrier Proteins↗