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Articles by H. Higashino
Total Records ( 5 ) for H. Higashino
  Tz. Markova , M. Tabuchi , B. Alexieva , E. Nikolova , Y. Aragane and H. Higashino
  Phosphodiesterase (PDE) enzymes catalyze breakdown of cAMP and cGMP, which is known to have potent immunomodulatory capacity. Here, we aimed at clarifying whether PDE4 is involved in allergen-induced activation of immunocompetent cells. To address this issue, human peripheral blood-derived mononuclear cells (PBMC) were cultured with mite antigen, dermatophagoides farinae (Der-f), in the presence or the absence of a specific PDE4 inhibitor, rolipram or a non-selective PDE inhibitor theophylline. Semiquantitative RT-PCR analysis demonstrated that stimulation of cells with Der-f led to upregulated expression of IL-5 mRNA, which was clearly downregulated by both inhibitors. Downregulation of Der-f-induced IL5 mRNA was similarly observed when cells were treated with U0126, a specific MEK/ERK inhibitor. This suggests that ERK signaling pathways are involved in this event. This was further supported by the observation that rolipram as well as theophyllin interfered with phospholylation of ERK1/2 induced by Der-f as determined by Western blot analysis using an antibody directed against phospholylated ERK1/2. Finally, immunostaining showed that stimulation to PBMC with Der-f led to nuclear translocation of AP-1 components (JunD, Fra-2), whose migration was inhibited by rolipram, indicating the interference with activation of AP-1 transcription factors. Together, the present study indicates that allergen-induced activation of immunocompetent cells was prevented by rolipram through interference with ERK1/2 signaling.
  H. Higashino , M. Tabuchi , S. Yamagata , T. Kurita , H. Miya , H. Mukai and Y. Miya
  Since Nitric Oxide (NO) is produced by three types of Nitric Oxide Synthases (NOSs), rapid changes in stable oxidized metabolites (nitrite and nitrate, NOx) in the tissues and blood should be represented by the amount of stable forms in serum and may reflect vascular activities and circulatory or inflammatory changes in the body. Therefore, serum NOx levels in patients with various diseases were measured and compared to healthy controls. Four hundred and sixty five in- and outpatients aged 14 to 96 years were included in this study and 49 healthy hospital workers were included in the control group. The NOx levels of both groups were measured at rest in the morning using an ozone chemiluminescence method. When compared with the control group, serum NOx levels were higher in patients consist of around 40 or more in numbers diagnosed with cardiovascular diseases including myocardial infarction, hyperlipaemia, gastrointestinal diseases including acute enteritis, chronic liver diseases including viral B and C type hepatitis and liver cirrhosis, diabetes in males, hypertension with comorbid diseases. In addition, patient groups with renal disorders, hyperuricemia, osteoporosis, untreated cancers, autoimmune diseases such as Rheumatoid Arthritis (RA) and Systemic Lupus Erythematosus (SLE) were also higher, although the each number might be not enough to compare between the groups. Patient groups with acute and chronic inflammation had significantly higher serum NOx levels. Therefore, measurement of serum NOx in patients may be useful for understanding the status and pathophysiology of inflammatory diseases and those in which inflammation is a component.
  H. Endo , M. Tabuchi , M.S. Ashenagar , K. Ooshima , H. Chen and H. Higashino
  To clarify the role of adrenal glands in hypertensive animals, levels of catecholamines and corticosteroids in plasma and the mRNA expression of the associated enzymes were measured in 6 and 9-week-old stroke-prone spontaneously hypertensive rats (SHRSP) and normotensive Wistar Kyoto rats (WKY) following cold stress. With and without cold stress immersing in cold water at 4°C for 15 sec, catecholamines, adrenocorticotropic hormone (ACTH) and corticosteroids in plasma and mRNA expression in adrenal glands were measured using high performance liquid chromatography (HPLC), enzyme immunoassay (EIA) and DNA microarray, respectively. L-dopa, dopamine and adrenaline in plasma increased more in SHRSP than WKY at 6 and 9 weeks of age after cold stress. Th, Ddc and Dbh mRNAs were upregulated in the adrenal glands of SHRSP after cold stress, more apparent at 6 weeks than at 9 weeks of age. Corticosterone and aldosterone in plasma increased in both SHRSP and WKY, but this effect was more apparent in SHRSP after elevation of ACTH evoked by cold stress. Expressions of cyp11a1 and cyp21a1 mRNAs were upregulated in both SHRSP and WKY at 6 weeks of age after cold stress. We conclude that l-dopa, dopamine, and adrenaline were synthesized following induction of Th, Ddc and Dbh mRNAs. Corticosterone and aldosterone in plasma increased following the induction of cyp11a1 and cyp21a1 mRNAs which are stimulated along with ACTH elevation following cold stress in young SHRSP more than WKY. This difference may be related to the initiation and/or development of hypertension in SHRSP in normal condition and/or during stress.
  T. Kurita , K. Nakamura , M. Tabuchi , M. Orita , K. Ooshima and H. Higashino
  The details of pharmacological mechanisms of Gorei-san, a traditional Japanese Kampo medicine, remains to be clarified, although it has been used for diuretic and hydrostatic purposes. From these circumstances, the effects of this medicine on the expressions of aquaporin (AQP) 1, 2, 3, 4 and vasopressin 2 receptor (V2R) mRNAs were investigated in relation to diuresis and water balance regulation in the kidney and brain. Gorei-san extract decocted with hot water was given to rats loaded with 50 mL kg-1 volume of physiological saline and AQP1, 2, 3, 4 and V2R mRNAs were measured with real-time polymerase chain reaction (PCR) in the cortex and the medulla of kidney and the forebrain. A low dose of Gorei-san extract (100 mg kg-1) led to an increase in urine excretion and lower AQP3 mRNA expression in the cortex as well as lower expression of AQP2 and AQP3 mRNAs in the medulla of kidney, whereas no change in V2R mRNA expression was observed. AQP1 mRNA expression decreased in the forebrain of rats loaded with an excess volume of physiological saline compared with rats not loaded with excess saline and given no agent. A low dose of Gorei-san extract increased urine excretion volume, probably due to the downregulation of AQP3 mRNA in the cortex and downregulation of AQP2 and AQP3 mRNAs in the medulla of the kidney, in which changes were not related to V2R mRNA expression. An excess volume of physiological saline given to rats caused an inhibition of AQP1 mRNA expression in the forebrain, which probably functioned to maintain the water balance in a hyper-hydrous state.
  T.P. Markova , Y. Aragane , A. Niwa , M. Tabuchi , M. Orita , K. Ooshima and H. Higashino
  Phosphodiesterases (PDE) catalyze the breakdown of cAMP and thus, may have potent immunomodulatory activity. Theophyllin is a non-selective PDE inhibitor and has long been used to treat atopic diseases such as asthma, based on its well-known effects of the bronchodilative and anti-inflammatory effects, although the immune modulatory activities are not completely clear yet. Because it is believed that Th2-immnuity plays a pathogenic role in asthma, we were interested to study the mechanisms of how it modulates mite-allergen-induced activation of Th2-like immunity. To address this issue, peripheral blood mononuclear cells (PBMC) were prepared from human healthy volunteers, stimulated with mite-allergen dermatophagoides farinae (Der-f), RNA prepared and semi-quantitative RT-PCR was performed using primers specific for IL-5, a representative Th2 cytokine. Consequently, it was found that theophylline at a concentration of 10-6 to 10-5 M significantly downregulated Der-f-induced expression of IL-5 mRNA. Intriguingly, theophylline (10-5 M) inhibited the expression of IL-5 mRNA of PBMCs from patients allergic to house dust mite. To further explore the mechanisms involved, a PKA type I inhibitor Rp-8-Br-cAMPS was added, resulting in restoration of theophylline-downregulated expression of Der-f-induced IL-5 mRNA, indicating the involvement of PKA in this event. Together, the present study demonstrates that theophylline prevents allergen-induced PKA activation and thus may, at least in part, inhibit Th2-like immune responses.
 
 
 
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