Biomedical subjects

K Momose

Publications and source records attributed to K Momose.

At least 19 recordsLinked to original sources

PubMed1996

Lysophosphatidic acid sensitizes mechanical stress-induced Ca2+ mobilization in cultured human lung epithelial cells.

We conformed that lysophosphatidic acid (LPA), which is known to be released from activated platelets, sensitizes response in cytosolic free Ca2+ concentration ([Ca2+]i) to mechanical stimulation in cultured epithelial cells (REPF-LC-AI cells) from human lung carcinoma. [Ca2+]i was transiently increased by spritzing of bath solution onto cells as mechanical stimulation in the presence of LPA with concentration-dependent manner (10-100 nM). The transient increase induced by the mechanical stimulation in the presence of LPA was inhibited by 10 microM Ga3+ or removing extracellular Ca2+, but not by 10 microM nicaridipine, suggesting that LPA sensitizes mechanical stimulation-induced Ca2+ influx through stretch-activated ion channels. Phosphatidic acid (1 microM), but not lysophosphatidycholine (10 microM), histamine (100 nM), bradykinin (10 nM), nor ionomycin (100 nM), caused the same effect as that of LPA. This effect was observed in confluent cells, but not in subconfluent cells. These results show that LPA sensitizes mechanoreceptor-linked response in human lung epithelial cells, suggesting a possibility that LPA affects lung function, in particular, during pathological state.

Calcium↗

PubMed1996

The requirement for and mobilization of calcium during induction by sodium ascorbate and by hydrogen peroxide of cell death.

The requirement for and mobilization of Ca2+ ions during induction of cell death by sodium ascorbate were compared with those during induction of cell death by hydrogen peroxide. When HL-60 cells were incubated with sodium ascorbate, a rapid increase in the intracellular concentration of Ca2+ ions and subsequent apoptotic cell death, characterized by cell shrinkage, nuclear fragmentation and cleavage of internucleosomal DNA to yield fragments that were multiples of 180-200 base pairs, were induced. However, these effects of sodium ascorbate were significantly reduced in Ca2+-depleted medium. By contrast, hydrogen peroxide induced similar apoptosis associated phenomena in the presence and in the absence of extracellular Ca2+ ions. The intracellular concentration of the reduced form of glutathione was not significantly affected and glutathione disulfide was undetectable during the early stages of apoptosis. These data suggest that sodium ascorbate and hydrogen peroxide initiate cell death by different mechanisms.

Apoptosis↗

PubMed1996

Functional coupling of the Na+/Ca2+ exchanger with Ca2+ release from intracellular stores in cultured smooth muscle cells of guinea pig ileum.

The mechanism of increase in intracellular Ca2+ concentration ([Ca2+]i) by removal of extracellular Na+, which phenomena were reported previously (Japan. J. Pharmacol. 63 83-91 1993), was investigated in cultured guinea pig ileum longitudinal muscle cells loaded with a fluorescent Ca2+ indicator, fura-2, by digital ratio imaging microscopy. Isotonic substitution of choline chloride for NaCl induced a transient increase in [Ca2+]i. The pretreatment of thapsigargin (0.5 microM), but not nicardipine (10 microM), suppressed the transient increase completely. In solutions containing micromolar concentrations of free Ca2+ (nominally Ca2+-free solution), the Na+-free induced transient increase was observed, but neither the second cell exposure to the Na+-free solution nor the following application of histamine increased [Ca2+]i, indicating that removal of extracellular Na+ releases Ca2+ from intracellular stores including inositol 1,4,5-trisphosphate (IP3)-releasable pools. The Na+-free induced transient increase required the presence of more than micromolar concentrations of extracellular free Ca2+ and releasable Ca2+ within the stores, but ryanodine did not affect the transient increase. These results suggest that undetectable influx of Ca2+ by the reverse-mode action of the Na+/Ca2+ exchanger can release Ca2+ from the thapsigargin-sensitive intracellular stores including IP3-releasable pools.

Animals↗

PubMed1996

Mechanisms of mechanical stress-induced Ca(2+)-mobilization sensitized by lysophosphatidic acid in cultured smooth muscle cells.

We have previously reported that lysophosphatidic acid (LPA) sensitizes mechanical stress-induced cytosolic free Ca2+ concentration ([Ca2+]i) response related to Ca2+ entry through Gd(3+)-sensitive ion channels (Biochem. Biophys. Res. Commun. 208 19-25 1995). Here we examined the contribution of Ca2 release from intracellular stores to the mechanical stress-induced [Ca2+]i response sensitized by LPA in cultured longitudinal muscle cells from guinea pig ileum. Although the percentage of responsive cells to the mechanical stress in the presence of 30 nM LPA declined by decreasing extracellular Ca2+ concentration to less than 20 microM, the amplitude of the mechanical stress-induced [Ca2+]i transient did not depend on extracellular Ca2+ concentrations (10 microM-1.8 mM). The [Ca2+]i transient was completely abolished by treatment with thapsigargin. In addition, the amplitude of the [Ca2+]i transient gradually decreased after ryanodine and caffeine treatment. These results indicate that the mechanical stress-induced [Ca2+]i transient in the presence of LPA is mainly due to Ca2+ release from ryanodine-sensitive intracellular stores and may be triggered by Ca2+ influx through Gd(3+)-sensitive ion channels.

Animals↗

PubMed1996

Sodium nitroprusside-induced apoptotic cellular death via production of hydrogen peroxide in murine neuroblastoma N1E-115 cells.

Sodium nitroprusside is widely used in pharmacological studies as a potent vasodilator or a nitric oxide donor. The mechanisms of cellular death induced by sodium nitroprusside were investigated in murine neuroblastoma N1E-115 cells. Sodium nitroprusside reduced the cellular viability, and the DNA extracted from treated cells showed a ladder-like intranucleosomal fragmentation pattern, which is an indication of apoptosis. The DNA fragmentations were also visualized by in situ nick translation. The cellular death was attenuated by cycloheximide, indicating that ongoing protein synthesis was essential for the initiation of the degenerative response. However, other nitric oxide donors did not decrease the cellular viability. The nitric oxide scavenger, hemoglobin, had no effect on sodium nitroprusside-induced cellular death. Furthermore, sodium cyanide, which is formed by the metabolism of sodium nitroprusside, did not cause cellular death. On the other hand, hydrogen peroxide, another product of sodium nitroprusside metabolism, reduced the cellular viability and induced DNA fragmentation. In addition, the cell damage induced by sodium nitroprusside was enhanced by a medium without fetal bovine serum. In conclusion, we proposed that hydrogen peroxide is the important toxic species for induction of apoptosis in N1E-115 cells exposed to sodium nitroprusside.

Animals↗

PubMed1996

A nitric oxide-sensitive electrode: requirement of lower oxygen concentration for detecting nitric oxide from the tissue.

In order to directly detect nitric oxide (NO) liberated from isolated tissue, a practical and convenient method using a nitric oxide-sensitive electrode is described. To avoid the nonselective signal caused by ionic substances, the electrode was covered with three layers but remains permeable for gaseous substances. In a solution bubbled with 20% oxygen (pO2, approximately 150 mm Hg), administration of S-nitroso-N-acetyl-d, l-penicillamine (SNAP) at concentrations greater than 10(-7) mol/L elicited an electrode response. Based on a comparison with the chemical determination of NO released from SNAP, the electrode may be able to detect nitric oxide around nmol/L. At least 30 nmol NO per liter in anoxic conditions was reported to be detected by this electrode (Matsui, 1995). In a specially designed small chamber, the electrode was attached on the surface of endothelial side of the isolated aorta of the guinea pig. When carbachol was added to the chamber, the electrode responded when the solution was bubbled with 20% but not with 40% or 95% of oxygen, suggesting a much faster decomposition of nitric oxide in the presence of higher concentrations of oxygen. The electrode response to carbachol was abolished in the presence of NG-monomethyl-L-arginine or nitro arginine. These results suggest that the electrode method described in this manuscript is suitable for detecting nitric oxide liberated from isolated tissues when comparatively low oxygen levels are present in the physiological salt solution.

Animals↗

PubMed1995

[3H]9-Methyl-7-bromoeudistomin D, a caffeine-like powerful Ca2+ releaser, binds to caffeine-binding sites distinct from the ryanodine receptors in brain microsomes.

[3H]9-Methyl-7-bromoeudistomin D ([3H]MBED), the most powerful Ca2+ releaser from sarcoplasmic reticulum, specifically bound to the brain microsomes. Caffeine competitively inhibited [3H]MBED binding. [3H]MBED binding was markedly blocked by procaine, whereas that was enhanced by adenosine-5'-(beta,gamma-methylene)triphosphate. The Bmax value was 170 times more than that of [3H]ryanodine binding. The profile of sucrose-density gradient centrifugation of solubilized microsomes indicated that [3H]MBED binding protein was different from [3H]ryanodine binding protein. These results suggest that there are MBED/caffeine-binding sites in brain that are distinct from the ryanodine receptor and that MBED becomes an essential molecular probe for characterizing caffeine-binding protein in the central nervous system.

Adenosine Triphosphate↗

PubMed1995

Protein kinase C is involved in translocation of diacyglycerol kinase induced by carbachol in guinea pig taenia coli.

The regulatory mechanisms of diacylglycerol (DG) kinase activity were studied in guinea pig taenia coli. In an octylglycoside mixed micellar assay system, DG kinase activities were distributed in both membrane and cytosolic fractions. Treatment of the tissue with carbachol (CCh) increased the activity in the membrane fraction and decreased the cytosolic fraction without affecting total DG kinase activity. The Km value of DG kinase in the membrane fraction was unchanged by treatment with CCh, although Vmax was increased. These findings suggest that DG kinase may be translocated from the cytosol to the membrane by CCh-stimulation. Increase in DG content by treatment of tissue with a cell-permeable species of DG, dioctanoyl-sn-glycerol, did not induce DG kinase translocation. Each treatment with protein kinase C (PKC) inhibitor and PKC-desensitization blocked CCh-induced DG kinase translocation; and phorbol ester induced the translocation only in intracellular calcium-accumulated tissues. Considering these results, CCh-induced DG kinase activation appears to involve DG kinase translocation from the cytosol to the membrane in association with both PKC and intracellular calcium concentration rather than cellular DG content.

Animals↗

PubMed1995

Sensitizing effect of lysophosphatidic acid on mechanoreceptor-linked response in cytosolic free Ca2+ concentration in cultured smooth muscle cells.

We found that lysophosphatidic acid (LPA) sensitizes response in cytosolic free Ca2+ concentration ([Ca2+]i) to mechanical stimulation in cultured longitudinal muscle cells from guinea pig ileum. [Ca2+]i was transiently increased by spritzing of bath solution onto cells as mechanical stimulation in the presence of LPA, but not in absence of LPA. The effect was reversible and concentration-dependent (1-30 nM). Ga3+ but not nicardipine inhibited the [Ca2+]i transient in the presence of LPA. Phosphatidic acid also induced the sensitization, but the effective concentration was more than 10 times higher than in LPA. Histamine and carbachol did not have any sensitizing effect to mechanical stimulation. These results show that LPA sensitizes mechanoreceptor-linked response, suggesting that LPA may play an important role in mechanotransduction mechanisms as an endogenous regulatory factor.

Animals↗

PubMed1995

Pharmacological characterization of SR 48692 sensitive neurotensin receptor in human pancreatic cancer cells, MIA PaCa-2.

Previously, one type of human neurotensin receptor has been molecularly cloned. Recently, it has been proposed that a subtype of neurotensin receptor exists, which is not sensitive to newly synthesized neurotensin receptor antagonist SR 48692. In this study, we characterize the pharmacological properties of neurotensin receptor expressed in human pancreatic cancer cells, MIA PaCa-2. In binding studies with [3H]neurotensin, the data fit a model for a single population of high-affinity binding sites that are competitively antagonized by SR 48692. The rank order of the equilibrium dissociation rate constants for neurotensin(8-13), neurotensin, neuromedin N, [Ala11] neurotensin(8-13), and SR 48692 were similar to those found with the molecularly cloned human neurotensin receptors. Additionally, the intracellular calcium mobilization and the growth of MIA PaCa-2 cells induced by neurotensin receptor agonist were completely inhibited by SR 48692. In conclusion, our results showed that MIA PaCa-2 cells express functional and SR 48692-sensitive-type neurotensin receptors. It is suggested that the neurotensin receptor antagonist SR 48692 may be useful in the treatment of pancreatic cancers that possess this type of neurotensin receptor.

Calcium↗

PubMed1994

Activation of diacylglycerol kinase by carbachol in guinea pig taenia coli.

Changes in diacylglycerol kinase (DG kinase) activity in carbachol (CCh)-stimulated guinea pig taenia coli were investigated. In a mixed micellar assay system, added 1,2-dioctanoyl-sn-glycerol (diC8) and endogenous DG were competitively bound to common DG kinase isozymes from guinea pig taenia coli and phosphorylated, suggesting that diC8 is useful as a probe of agonist effects on DG kinase activity. In phosphorus-32 ([32P]Pi)- and diC8-prelabeled guinea pig taenia coli, diC8 was phosphorylated by DG kinase to [32P]dioctanoyl-phosphatidic acid ([32P]diC8-PA). CCh increased the accumulation of both [32P]diC8-PA and endogenous [32P]phosphatidic acid ([32P]PA) in a time- and dose-dependent manner (0.1-100 microM CCh). CCh-induced increases in [32P]diC8-PA and [32P]PA were inhibited by 1 microM atropine and 3 microM DG kinase inhibitor (R59022). These findings indicated the activation of DG kinase by muscarinic receptor stimulation in guinea pig taenia coli. Therefore, DG kinase activation may play an important role in CCh-induced PA formation. CCh-induced [32P]diC8-PA and [32P]PA accumulation was dependent on intracellular calcium concentrations. However, a KCl-induced increase in intracellular calcium, without receptor stimulation, was ineffective. Moreover, treatment with phorbol ester also increased accumulation of both PA species in KCl-treated tissues. These findings suggest that muscarinic receptor mediated activation of DG kinase may require both an increase in intracellular calcium and PKC activation in guinea pig taenia coli.

Animals↗

PubMed1994

3',3",5',5"-Tetraiodophenolsulfonephthalein is a selective inhibitor of Ca(2+)-pumping ATPase in intracellular Ca2+ store.

3',3",5',5"-Tetraiodophenolsulfonephthalein (I4PSP) stimulated both Ca2+ transient levels and aggregation in response to thrombin in platelets. Ca2+ uptake into skeletal sarcoplasmic reticulum (SR) was inhibited by I4PSP (IC50, 1.8 microM) whereas that of red blood cell ghosts was not affected by it. Furthermore, I4PSP inhibited the SR Ca(2+)-ATPase activity (IC50, 1.1 microM). Kinetic analysis of the inhibitory effects of I4PSP reveals that I4PSP shows an inhibition of noncompetitive and competitive type with respect to low and high concentrations of ATP, respectively. The mode of inhibition by I4PSP is an uncompetitive type with respect to Ca2+. I4PSP decreased the decomposition rate of the phosphorylated enzyme intermediate (EP). The change in the tryptophan fluorescence of SR Ca(2+)-ATPase induced by ATP was reduced by I4PSP indicating inhibition of the EP transition from Ca2E1P to E2P. The concentration-inhibitory response curves for I4PSP in Ca(2+)-ATPase activities and fluorescence changes were closely correlated. These results suggest that I4PSP slows down the structure transition from Ca2E1P to E2P, resulting in inhibition of the Ca(2+)-ATPase activity. On the basis of these results, it is suggested that the stimulation of Ca2+ transient levels in platelets and their aggregation in response to thrombin are due to the inhibition of Ca2+ pump in intracellular Ca2+ stores by I4PSP.

Adenosine Triphosphate↗

PubMed1994

Reduction by NG-nitro-L-arginine of H2O2-induced endothelial cell injury.

1. The effects of three analogues of NG-nitro-L-arginine (L-NOARG) and NG-monomethyl-L-arginine (L-NMMA), inhibitors of nitric oxide (NO) synthase, on hydrogen peroxide (H2O2)-induced endothelial cell injury were studied. 2. Endothelial cell injury was assessed by measuring the release of intracellular lactate dehydrogenase (LDH) and 51Cr. 3. Addition of H2O2 (250-1,000 microM) to endothelial cells induced the release of LDH dose-dependently. The release of LDH was reduced by pretreatment with NG-nitro-L-arginine methyl ester (L-NAME, 10(-4)-4 x 10(-3) M), L-NOARG (10(-4)-4 x 10(-3) M) and NG-nitro-L-arginine benzyl ester (L-NABE, 10(-4)-4 x 10(-3) M), inhibitors of NO synthase. 4. L-NOARG analogues also reduced H2O2-induced 51Cr release from endothelial cells, while L-NMMA had no effect. 5. The protective effect of L-NAME was not reversed by addition of L-arginine (L-Arg, 1-10 mM). 6. Both L-NAME and L-NMMA completely inhibited L-Arg metabolism to L-citrulline coupled with NO synthesis. 7. These findings suggest that L-NOARG analogues but not L-NMMA reduced H2O2-induced endothelial cell injury, and that these effects may not be related to inhibition of NO production.

Amino Acid Oxidoreductases↗

PubMed1994

Turbidimetric latex agglutination inhibition immunoassay for primidone.

We have developed a quick, highly sensitive immunoassay method for drugs by latex agglutination inhibition. An antiserum against primidone (PRM) was obtained by immunizing rabbits with PRM-bovine serum albumin conjugate. PRM-rabbit serum albumin conjugate sensitized latex was agglutinated with diluted antiserum, and the agglutination was inhibited by free PRM quantitatively. Turbidity of the agglutination suspension was measured by spectrophotometry as absorbance. Larger latex gave higher sensitivity than the smaller, because its agglutination was inhibited more intensely by free PRM. The assay values of this method were correlated well with those obtained by an enzyme immunoassay method.

Animals↗

PubMed1994

The relationship between spontaneous calcium oscillations and cell proliferation in cultured smooth muscle cells.

The relationship between spontaneous Ca2+ oscillations and cell proliferation was investigated in cultured longitudinal muscle cells from guinea pig ileum. BAPTA, caffeine, thapsigargine and La3+ suppressed the spontaneous Ca2+ oscillations and the DNA synthesis at 5.0 mM, 10.0 mM, 15.62 nM and 0.5 mM, respectively, whereas verapamil, nicardipine and ryanodine did not suppress either. However, BAPTA and La3+ did not inhibit the DNA synthesis at 1.25 mM and 0.13 mM, but did suppress the Ca2+ oscillations even at 1.0 mM and 0.1 mM, respectively. These results show that the spontaneous Ca2+ oscillations are not prerequisite for the cell proliferation.

Animals↗

PubMed1994

Bradykinin induces generation of reactive oxygen species in bovine aortic endothelial cells.

We investigated the effects of bradykinin on intracellular oxidative stress in bovine aortic endothelial cells using a hydroperoxide-sensitive fluorescent dye, 2',7'-dichlorofluorescein (DCFH), and a laser scanning confocal microscope. Bradykinin induced an immediate increase in intracellular Ca2+ concentration, and stimulated the oxidation of DCFH in cultured endothelial cells. This bradykinin-induced oxidation of DCFH was inhibited by pretreatment with N-(2-mercaptopropionyl)-glycine (MPG) and 1,3-dimethyl-thiourea (DMTU), scavengers of hydroxyl radical, and the removal of extracellular Ca2+ but was unaffected by NG-nitro-L-arginine or NG-monomethyl-L-arginine, both inhibitors of nitric oxide (NO) synthase. On the other hand, pretreatment with indomethacin and aspirin, inhibitors of cyclooxygenase, inhibited bradykinin-induced oxidation of DCFH. These findings suggest that bradykinin increases intracellular Ca2+ and stimulates the generation of hydroxyl radical-like reactive oxygen species (scavenged by MPG or DMTU) via the cyclooxygenase pathway but not via the reaction of NO and superoxide anion.

Amino Acid Oxidoreductases↗

PubMed1994

All-or-none like responses in increment of cytoplasmic free calcium concentrations to histamine in single smooth muscle cells of guinea pig trachea.

Single smooth muscle cells were isolated from guinea pig trachea and cultured for 48 hours. The cells were then loaded with a fluorescent-free calcium indicator, fluo-3. Concentrations of cytoplasmic-free calcium ([Ca2+]i) were determined by using a confocal laser scanning fluorescence imaging system. Cells exposed to histamine at concentrations less than 1.0 microM did not indicate any changes of the fluorescence intensities, but stayed at the basal level of [Ca2+]i. Cells exposed to histamine at a concentration in the range of 1.5 to 10 microM indicated marked increases in [Ca2+]i. A further increase in [Ca2+]i by application of higher concentrations of histamine was not observed. The threshold concentration of histamine to increase [Ca2+]i and peak [Ca2+]i varied from cell to cell. In addition, half-maximal response time was shortened with increasing concentrations of histamine from 1.5 to 100 microM. These results indicate that [Ca2+]i responses to histamine is an all-or-none type in each cell. The heterogeneity in the required threshold concentration of histamine to increase in [Ca2+]i and the concentration dependency in half-maximal response time of histamine-induced [Ca2+]i increase may be related to the graded responses of histamine-induced contractions in preparations of the tracheal tissue.

Aniline Compounds↗

PubMed1994

Stimulation by hydrogen peroxide of L-arginine metabolism to L-citrulline coupled with nitric oxide synthesis in cultured endothelial cells.

The effect of hydrogen peroxide on metabolism of L-arginine to L-citrulline in the biosynthesis of nitric oxide was investigated in bovine aortic endothelial cells. Addition of hydrogen peroxide (1-10 mM) to endothelial cells increased L-citrulline formation from L-arginine in a dose-dependent manner. Stimulation of L-citrulline formation was prevented by catalase (20 micrograms/ml), a hydrogen peroxide scavenger, but not by superoxide dismutase (SOD, 100 micrograms/ml), a superoxide anion scavenger. Pretreatment with NG-nitro-L-arginine (10(-5) M) and NG-methyl-L-arginine (10(-4) M), potent inhibitors of nitric oxide synthase, also inhibited L-citrulline formation induced by hydrogen peroxide. Moreover, hydrogen peroxide increased intracellular Ca2+ concentrations, and the removal of extracellular Ca2+ reduced the hydrogen peroxide-induced formation of L-citrulline. These findings show that hydrogen peroxide increases the intracellular Ca2+ concentration and stimulates the formation of L-citrulline from L-arginine coupled with nitric oxide synthesis in cultured endothelial cells.

Animals↗