Journal of Pharmacological Sciences
Online ISSN : 1347-8648
Print ISSN : 1347-8613
ISSN-L : 1347-8613
Volume 95 , Issue 4
Showing 1-12 articles out of 12 articles from the selected issue
Full Papers
  • Toyohisa Hanano, Yuji Hara, Juan Shi, Hiromitsu Morita, Chisato Umebay ...
    2004 Volume 95 Issue 4 Pages 403-419
    Published: 2004
    Released: August 21, 2004
    [Advance publication] Released: July 24, 2004
    JOURNALS FREE ACCESS
    We investigated the possible involvement of the melastatin family protein TRPM7 in Ca2+-mediated proliferative control of human retinoblastoma (RB) cells. The growth of RB cell was facilitated by elevating the extracellular Ca2+ concentration with a parallel increase in the magnitude of spontaneous Ca2+ influx. Under nystatin-perforated voltage-clamp, RB cells exhibited an outward-rectifying, spontaneous cation current (Ispont) having Ca2+/Mg2+-inhibited but -permeating properties. Various cation channel blockers inhibiting Ispont (Gd3+, La3+, LOE908, 2-APB) suppressed the spontaneous Ca2+ influx and decelerated the growth of RB cells with similar efficacies. Excision of the RB cell membrane (inside-out) into MgATP-free solution induced a 70pS single channel activity, which was effectively inhibited by millimolar concentrations of Mg2+ or MgATP. RT-PCR and immunocytochemical experiments revealed the expression of TRPM7 mRNA and protein in RB cells, and heterologous expression of TRPM7 in HEK293 cells reproduced the key features of Ispont. In contrast, elimination of this protein from RB cells by siRNA silencing markedly reduced Ispont density and the magnitude of spontaneous Ca2+ influx, which was paralleled by decreased TRPM7 immunoreactivity, decelerated cell proliferation, and retarded G1/S cell cycle progression. These results suggest a significant regulatory role of TRPM7 for RB cell proliferation as a spontaneously activated Ca2+ influx pathway.
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  • Tsutomu Sakuma, Xiu Gu, Makoto Sugita, Motoyasu Sagawa, Maki Sakuda, H ...
    2004 Volume 95 Issue 4 Pages 420-425
    Published: 2004
    Released: August 21, 2004
    [Advance publication] Released: July 24, 2004
    JOURNALS FREE ACCESS
    Uridine 5'-triphosphate (UTP) increases chloride secretion followed by fluid movement into the proximal airspaces. However, little is known about whether UTP affects fluid movement in the distal airspaces. We studied the effect of UTP on basal and stimulated alveolar fluid clearance in the isolated rat lungs. Isosmotic 5% albumin solution was instilled into the alveolar spaces of isolated rat lungs, which were then inflated with 100% oxygen at an airway pressure of 7 cmH2O. Alveolar fluid clearance was measured by the progressive increase in albumin concentrations over 1 h. Although UTP (109 – 106 M) did not increase alveolar fluid clearance, UTP (105 – 103 M) and isoproterenol (105 M), a β-adrenergic agonist, increased alveolar fluid clearance by 40% and 120% of the basal values, respectively. A combined treatment of UTP (104 M, 103 M) and isoproterenol increased alveolar fluid clearance by 280% of the basal value. The effects of UTP in the presence and absence of isoproterenol were abolished by blockers of a P2 purinoceptor and chloride channels. These results indicate that UTP stimulates alveolar fluid clearance in the distal airspaces of rat lungs.
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  • Katsuhiro Miyoshi, Nozomi Kawakami, Yousuke Wakayama, Norimasa Izumi, ...
    2004 Volume 95 Issue 4 Pages 426-434
    Published: 2004
    Released: August 21, 2004
    [Advance publication] Released: July 24, 2004
    JOURNALS FREE ACCESS
    Heterologous down-regulation of histamine H1 receptor (H1R) mediated by muscarinic acetylcholine receptor subtype was investigated using five kinds of Chinese hamster ovary (CHO) cells stably co-expressing the human H1R and one of the five (M1 – M5) muscarinic acetylcholine receptors, CHO-H1/M1, CHO-H1/M2, CHO-H1/M3, CHO-H1/M4, and CHO-H1/M5 cells. Among the CHO-H1/M1, CHO-H1/M3, and CHO-H1/M5 cells, carbachol treatment of the CHO-H1/M3 cells time-dependently led to remarkable down-regulation of the H1R to 60% of the control level. In contrast, stimulation of CHO-H1/M1 cells by carbachol induced negligible effect on the down-regulation. Stimulation of CHO-H1/M5 cells by carbachol induced significant but only small H1R down-regulation. M2 and M4 muscarinic receptors showed negligible effect on the down-regulation. H1R-mediated accumulation of inositol phosphates in CHO-H1/M3 cells with long-term expose to carbachol was decreased to 60% compared with non-treated cells. Heterologous phosphorylation of H1R was induced by the stimulation of each muscarinic receptor. H1R was phosphorylated by about twofold from the basal level through five subtypes of muscarinic receptor. The M3 muscarinic receptor-mediated phosphorylation of H1R was reversed by the inhibition of protein kinase C. In the present study we demonstrated that the M3 muscarinic acetylcholine receptor mediated remarkable down-regulation of the H1R with decreased receptor signaling.
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  • Masatomo Kato, Kenji Imoto, Hideki Miyake, Tomoko Oda, Suguru Miyaji, ...
    2004 Volume 95 Issue 4 Pages 435-442
    Published: 2004
    Released: August 21, 2004
    [Advance publication] Released: July 31, 2004
    JOURNALS FREE ACCESS
    Platelet-activating factor (PAF) plays important roles in allergic reactions. In particular, there are many concerns about PAF, eosinophils, and the chronicity of allergic diseases. The purpose of the present studies is to elucidate the role of PAF in eosinophil activation at conjunctiva and to confirm the efficacy of Apafant (a potent PAF antagonist) ophthalmic solution in chronic experimental allergic conjunctivitis. Guinea pigs were actively immunized and allergic conjunctivitis was induced by repetitive instillation of 2.5% ovalbumin. PAF solution was topically applied and eosinophil activation was assessed by measuring the eosinophil peroxidase (EPO) activity in the tear fluid. Itch-scratching episodes and clinical symptoms scores were evaluated in the repetitive challenge conjunctivitis. From the instillation of PAF solution into guinea pig eyes, which were in a state of chronic allergic conjunctivitis, a significant increase in EPO activity was observed, and this increase was inhibited by pre-treatment with Apafant. In the repetitive challenge model, the animals treated with Apafant ophthalmic solution showed a significant reduction of clinical symptoms and the itch-scratch response in both the first and the second challenges. PAF has an activity, that induces mediator release from eosinophils in the conjunctival tissues and may be involved in the chronic phase of allergic conjunctivitis.
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  • Mayumi Mori, Hiromi Tsushima
    2004 Volume 95 Issue 4 Pages 443-451
    Published: 2004
    Released: August 21, 2004
    [Advance publication] Released: July 31, 2004
    JOURNALS FREE ACCESS
    We characterized the effects of vanadate, an inhibitor of tyrosine phosphatase, on the tension, the level of myosin light chain (MLC) phosphorylation, and Rho A activation in intact ileal longitudinal smooth muscle of the guinea pig to study the role of tyrosine phosphorylation in contraction signaling. Vanadate exerted a sustained contraction with a slow onset of tension development, in a concentration-dependent manner. The contractile effects of vanadate were accompanied by increases in the level of MLC phosphorylation. The tyrosine kinase inhibitor genistein; the MLC kinase inhibitor 1-(5-chloronaphthalene-1-sulfonyl)-1H-hexahydro-1,4-diazepine hydrochloride (ML-9); and the Rho kinase inhibitor (+)-(R)-trans-4-(1-aminoethyl)-N-(4-pyridyl) cyclohexanecarboxamide dihydrochloride, monohydrate (Y-27632) inhibited the vanadate-induced contraction and MLC phosphorylation. Vanadate caused Rho A translocation from the cytosol to the membrane fraction, which was inhibited by genistein, but not by ML-9 and Y-27632. These data indicate that vanadate induces Rho A activation probably via protein tyrosine phosphorylation and the subsequent contraction through increases in the level of MLC phosphorylation.
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  • Norifumi Shimizu, Shiroh Kishioka, Takehiko Maeda, Yohji Fukazawa, Yos ...
    2004 Volume 95 Issue 4 Pages 452-457
    Published: 2004
    Released: August 21, 2004
    [Advance publication] Released: August 05, 2004
    JOURNALS FREE ACCESS
    Morphine’s analgesic actions are thought to be mediated through both the central and peripheral nervous systems. L-type calcium channel blockers have been reported to potentiate the analgesic effects of morphine, but the locus of this interaction is not known. In this experiment, we examined the site of verapamil-induced potentiation of morphine analgesia in mice using the quaternary opioid receptor antagonist naloxone-methiodide (NLX-M). Subcutaneous injections of morphine increased locomotor activity and serum corticosterone level, which are mediated by the central nervous system. These central effects were not antagonized by 0.1 mg/kg of NLX-M, whereas this dose of NLX-M partially antagonized the analgesic effect of morphine. Treatment with verapamil potentiated morphine analgesia in a dose-dependent manner. The verapamil-induced potentiation of morphine analgesia was abolished by pretreatment with NLX-M (0.1 and 1 mg/kg). These findings suggest that peripheral mechanisms partially contribute to morphine analgesia and mediate the potentiation of morphine analgesia by verapamil.
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  • Hiroko Takagi-Matsumoto, Betty Ng, Yasuhiro Tsukimi, Masaomi Tajimi
    2004 Volume 95 Issue 4 Pages 458-465
    Published: 2004
    Released: August 21, 2004
    JOURNALS FREE ACCESS
    To clarify the potential usefulness of non-steroidal anti-inflammatory drugs, NSAIDs, for patients with overactive bladder, we examined the effect of NSAIDs on urodynamic parameters in normal and cystitis rats and compared their ulcerogenic activity in the gastrointestinal mucosa. Cystometry was performed after administration of the conventional NSAIDs, aspirin, indomethacin, or ketoprofen. Prostaglandin levels were measured in the bladder of cystitis rats pretreated with NSAIDs. Furthermore, the ulcerogenic responses were examined. NSAIDs increased bladder capacity without any effect on micturition pressure in normal rats in the following rank order of potency: ketoprofen ≥ indomethacin ≥ aspirin. In cystitis rats, bladder capacity was increased and micturition frequency was decreased. The levels of prostaglandin were significantly increased in cystitis rats. All NSAIDs inhibited the increment of prostaglandin levels at doses equal to that effective in the improvement of bladder functions. When administered intraduodenally, both ketoprofen and indomethacin induced lesions in the gastrointestinal mucosa. However, aspirin had no significant effect. We demonstrate that NSAIDs are effective in animal models of disease, most likely by suppressing by prostaglandin synthesis. Since aspirin, in contrast to ketoprofen or indomethacin, did not cause any gastrointestinal lesions, aspirin might be the NSAIDs treatment of choice for overactive bladder.
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Short Communications
  • Mary Ann Suico, Zhuo Lu, Tsuyoshi Shuto, Tomoaki Koga, Tomoko Uchikawa ...
    2004 Volume 95 Issue 4 Pages 466-470
    Published: 2004
    Released: August 21, 2004
    [Advance publication] Released: July 31, 2004
    JOURNALS FREE ACCESS
    The human β-defensin 2 (HBD 2) is a potent anti-bacterial peptide with a wide spectrum of activity. MEF (myeloid elf-1-like factor) or Elf4, a member of the ETS transcription factor family, has been shown to up-regulate the basal expression of the HBD 2 gene in epithelial cells. The mammalian cell nucleus is organized into distinct nuclear domains, one of which is the PML (promyelocytic leukemia) nuclear body involved in the regulation of transcription. Here, we show that PML stimulated MEF transcriptional activity, resulting in the up-regulation of endogenous HBD 2 expression.
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  • Tsukasa Okiyoneda, Kazutsune Harada, Kaori Yamahira, Ikuo Wada, Yasuak ...
    2004 Volume 95 Issue 4 Pages 471-475
    Published: 2004
    Released: August 21, 2004
    [Advance publication] Released: July 31, 2004
    JOURNALS FREE ACCESS
    To examine the unknown trafficking pathway of the cystic fibrosis transmembrane conductance regulator (CFTR) from the endoplasmic reticulum (ER), we utilized baby hamster kidney cells stably expressing CFTR fused with green fluorescent protein. CFTR trafficking from the ER was visualized and analyzed by immunocytochemical analyses. Here we show that CFTR was exported from the ER to the cis-Golgi and early endosome, suggesting that CFTR transport in the early secretory pathway may utilize a non-conventional pathway. This CFTR trafficking pathway may be a target for pharmacological modulation that selectively stimulates CFTR transport.
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  • Mamiko Kai, Atsushi Yamauchi, Koji Tominaga, Ayako Koga, Hisashi Kai, ...
    2004 Volume 95 Issue 4 Pages 476-478
    Published: 2004
    Released: August 21, 2004
    [Advance publication] Released: August 05, 2004
    JOURNALS FREE ACCESS
    Hot flushes are one of the most frequent symptoms in menopausal women. We investigated effect of soybean isoflavones (Soyaflavone HG) on nifedipine-induced flushing in ovariectomized mice. Ovariectomy markedly aggravated nifedipine-induced increase in tail skin temperature. Soyaflavone HG (10 mg/kg, p.o., once a day for 5 days) inhibited nifedipine-induced flushing in ovariectomized mice. The inhibitory effect of Soyaflavone HG was significantly reversed by an estrogen-receptor antagonist, ICI 182,780, suggesting that Soyaflavone HG prevents nifedipine-induced flushing partially through estrogen receptors. We presented the experimental evidence suggesting that soybean isoflavones including Soyaflavone HG have the benefits for menopausal hot flushes.
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  • Yoshihiko Chiba, Tetsuro Uchida, Hiroyasu Sakai, Teruaki Oku, Saotomo ...
    2004 Volume 95 Issue 4 Pages 479-482
    Published: 2004
    Released: August 21, 2004
    [Advance publication] Released: August 05, 2004
    JOURNALS FREE ACCESS
    By using immunofluorostaining and confocal laser microscopy, acetylcholine-induced translocation of RhoA was visualized in freshly isolated bronchial smooth muscle cells of the rat. The cellular distribution of RhoA at rest was observed uniformly in the cytosolic space with no staining in the nucleus, whereas acetylcholine stimulation induced a relocalization of RhoA to the cell periphery. From the results of line scans and surface plots, the peripheral to cytosolic ratio of RhoA was significantly increased by acetylcholine stimulation. Thus, the present study clearly demonstrated an acetylcholine-induced translocation of RhoA to the plasma membrane in single bronchial smooth muscle cells of the rat.
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  • Moe Kyaw, Masanori Yoshizumi, Koichiro Tsuchiya, Yuki Izawa, Yasuhisa ...
    2004 Volume 95 Issue 4 Pages 483-486
    Published: 2004
    Released: August 21, 2004
    [Advance publication] Released: August 05, 2004
    JOURNALS FREE ACCESS
    We examined the effects of the stereoisomers of N-acetylcysteine (NAC), L-NAC and D-NAC, on cellular glutathione (GSH) concentration and whether NAC-regulated cellular GSH levels are directly associated with angiotensin II (Ang II)-induced intracellular signaling events in vascular smooth muscle cells (VSMC). Both L-NAC and D-NAC similarly increased intracellular GSH concentration. We found that L-NAC and D-NAC both inhibited Ang II-induced c-Jun N-terminal kinase and p38 mitogen-activated protein kinase activation and [3H]-thymidine incorporation in VSMC. Our present study indicates the comparable effects of NAC stereoisomers in regulating intracellular GSH and the redox-dependent intracellular signaling mechanisms in VSMC.
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