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Biomedical subjects

H Tokuda

Publications and source records attributed to H Tokuda.

At least 235 records · Page 13Linked to original sources
PubMed1993

Vasopressin induces arachidonic acid release through pertussis toxin-sensitive GTP-binding protein in aortic smooth muscle cells: independence from phosphoinositide hydrolysis.

We previously reported that pertussis toxin (PTX) had little effect on arginine vasopressin-induced formation of inositol trisphosphate (IP3) in rat aortic smooth muscle cells [Kondo et al.: Biochemical and Biophysical Research Communications 161:677-682, 1989]. In the present study, we investigated the mechanism of vasopressin-induced arachidonic acid release in rat aortic smooth muscle cells. Vasopressin stimulated both the release of arachidonic acid and the formation of IP3 dose dependently in the range between 10 pM and 1 microM. The effect of vasopressin on arachidonic acid release was more potent than that on the formation of IP3. Quinacrine, a phospholipase A2 inhibitor, significantly suppressed the vasopressin-induced arachidonic acid release but had little effect on the formation of inositol phosphates. NaF, a GTP-binding protein activator, mimicked vasopressin by stimulating the arachidonic acid release. The arachidonic acid release stimulated by a combination of vasopressin and NaF was not additive. PTX partially but significantly suppressed the vasopressin-induced arachidonic acid release. In the cell membranes, PTX catalyzed ADP-ribosylation of a protein with an M(r) of about 40,000. Pretreatment of membranes with 0.1 microM vasopressin in the presence of 2.5 mM MgCl2 and 100 microM GTP markedly attenuated this PTX-catalyzed ADP-ribosylation of the protein in a time-dependent manner. These results strongly suggest that PTX-sensitive GTP-binding protein is involved in the coupling of vasopressin receptor to phospholipase A2 in primary cultured rat aortic smooth muscle cells.

Adenosine Diphosphate Ribose↗
PubMed1993

Inhibitory effects of triptogelin A-1 on 12-O-tetradecanoylphorbol-13- acetate-induced skin tumor promotion.

Dihydroagarofuran sesquiterpenes, which were isolated from Tripterygium wilfordii Hook fil. var. regelii Makino and Euonymus sieboldianus Blume, showed inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by 12-O-tetradecanoylphorbol-13-acetate (TPA). Triptogelin A-1, one of the active dihydroagarofuran sesquiterpenes, exhibited remarkable inhibitory effects on mouse skin tumor promotion in an in vivo two-stage carcinogenesis test. Interestingly, triptogelin A-1 was more effective when applied after TPA-treatment (post-treatment) than before TPA-treatment (pretreatment). The findings indicate that these dihydroagarofuran sesquiterpenes might be valuable antitumor promoters.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1993

Effects of retinoic acid on signalling by prostaglandin E2 in osteoblast-like cells.

We investigated the effects of retinoic acid (RA) on the signalling pathways by prostaglandin E2 (PGE2) in osteoblast-like MC3T3-E1 cells. The pretreatment with RA significantly inhibited the formation of inositol phosphates induced by 10 microM PGE2 in a dose-dependent manner in the range between 0.1 nM and 0.1 microM, without affecting protein contents in the cultured cells. This effect of RA was dependent on the time of pretreatment up to 8 h. However, RA had little effect on the formation of inositol phosphates induced by NaF, a GTP-binding protein activator. On the other hand, RA significantly inhibited PGE2-induced cAMP accumulation in a dose-dependent manner between 0.1 nM and 0.1 microM. This effect of RA was dependent on the time of pretreatment up to 8 h. RA also inhibited the cAMP accumulation induced by NaF or forskolin which directly activates adenylate cyclase. These results strongly suggest that RA modulates the signalling by PGE2 in osteoblast-like cells as follows: the inhibitory effect on the phosphoinositide hydrolysis is exerted at the point between PGE2 receptor and GTP-binding protein, and the inhibitory effect on the cAMP production is exerted at a point downstream from adenylate cyclase.

3T3 Cells↗
PubMed1993

Modulation of prostaglandin E2-induced Ca2+ influx by steroid hormones in osteoblast-like cells.

In osteoblast-like MC3T3-E1 cells, we previously reported that prostaglandin E2 (PGE2), a potent bone resorbing agent, stimulates Ca2+ influx (H. Tokuda, M. Miwa, Y. Oiso and O. Kozawa, Cell Signal 1992; 4: 261-266). In this study, we examined the effects of various hormones belonging to the steroid hormone superfamily on PGE2-induced Ca2+ influx in MC3T-E1 cells. 1,25-Dihydroxyvitamin D3 [1,25-(OH)2D3], an active form of vitamin D3, dexamethasone and retinoic acid significantly inhibited the PGE2-induced Ca2+ influx in a dose-dependent manner in these cells. The effects of these hormones were dependent on the time of pretreatment and submaximum inhibitions were observed at 6 h. In contrast, 24,25-dihydroxyvitamin D3 [24,25-(OH)2D3], an inactive form of vitamin D3, 17 beta-estradiol, progesterone, testosterone and triiodothyronine had little effect on the PGE2-induced Ca2+ influx in these cells. These results suggest that, in the steroid hormone superfamily, 1,25-dihydroxyvitamin D3, glucocorticoid and retinoic acid modulate bone metabolism through the inhibition of Ca2+ influx induced by PGE2 in osteoblast-like cells.

Biological Transport↗
PubMed1993

Effects of glucocorticoid on signalling by prostaglandin E2 in osteoblast-like cells.

It is well-known that osteoporosis is a common complication of patients with glucocorticoid excess. We previously reported that dexamethasone inhibits Ca2+ influx induced by prostaglandin E2 (PGE2), a potent bone resorbing agent, in osteoblast-like MC3T3-E1 cells (O. Kozawa, H. Tokuda, J. Kotoyori, A. Suzuki, Y. Ito and Y. Oiso, Prostagland Leuk Essent Fatty Acids, in press). In the present study, we examined the effects of dexamethasone on cyclic adenosine monophosphate (cAMP) accumulation and phosphoinositide hydrolysis by PGE2 in MC3T3-E1 cells. The pretreatment with dexamethasone significantly inhibited cAMP accumulation stimulated by PGE2 in a dose-dependent manner in the range between 10 pM and 1 nM in MC3T3-E1 cells. This effect of dexamethasone was dependent on the time of pretreatment up to 8 h. Dexamethasone also inhibited the cAMP accumulation induced by NaF, an activator of guanosine triphosphate (GTP)-binding protein, or forskolin which directly activates adenylate cyclase. In contrast, dexamethasone had little effect on the formation of inositol phosphates stimulated by PGE2 in MC3T3-E1 cells. These results strongly suggest that glucocorticoid modulates the signal transduction by PGE2 in osteoblast-like cells and that it inhibits PGE2-induced cAMP production without affecting PGE2-induced phosphoinositide hydrolysis.

1-Methyl-3-isobutylxanthine↗
PubMed1993

Structures of sesquiterpene polyol esters from Celastrus stephanotiifolius with potential tumor-promotion inhibitor activity.

Esters of eight new ([1],[4-6], and [8-11]) and five known sesquiterpenenoid polyalcohols have been isolated from Celastrus stephanotiifolius. Their structures were established on the basis of chemical reactions and spectral analysis. The structural elucidation indicated that the structures of some related compounds should be revised. The isolated sesquiterpenes were observed to inhibit Epstein-Barr virus early antigen activation significantly at low doses.

Anticarcinogenic Agents↗
PubMed1993

Studies on inhibitors of skin tumor promotion, XII. Rotenoids from Amorpha fruticosa.

As a part of screening studies for chemopreventive agents (anti-tumor-promoters), six North American plants belonging to the Amorpha genus were tested using an in vitro assay system. Of these plants, Amorpha fruticosa exhibited strong inhibitory effects on Epstein-Barr virus early antigen (EBA-EA) activation induced by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA). Also six rotenoids, amorphispironone [1], tephrosin [2], amorphigenin [3], 12a-hydroxyamorphigenin [4], 12a-hydroxydalpanol [5], and 6'-O-D-glucopyranosyldalpanol [6], were isolated from the leaves of A. fruticosa. Among these retenoids, 1 and 2 exhibited remarkable inhibitory effects of EBV-EA activation induced by TPA. Further, 1 and 2 exhibited significant anti-tumor-promotion effects on mouse skin tumor promotion in an in vivo two-stage carcinogenesis test. These investigations suggested that these rotenoids might be valuable anti-tumor-promoters.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1993

Cross-talk between Ca2+ influx and phosphoinositide hydrolysis by extracellular adenosine triphosphate in rat PC12 pheochromocytoma cells.

In rat PC12 pheochromocytoma cells, extracellular ATP stimulated 45Ca2+ influx even in the presence of nifedipine, a Ca2+ antagonist that inhibits the voltage-dependent L-type Ca2+ channel, and stimulated the formation of inositol trisphosphate (IP3). The effect of ATP on 45Ca2+ influx was more potent than that on the formation of IP3 at a dose lower than 0.1 mM. 12-O-Tetradecanoylphorbol-13-acetate, a protein kinase-C (PKC)-activating phorbol ester, which by itself had little effect on 45Ca2+ influx, significantly reduced the ATP-induced 45Ca2+ influx in a dose-dependent manner in the range between 1 nM and 0.1 microM. However, 4 alpha-phorbol 12, 13-didecanoate, a PKC-nonactivating phorbol ester, had little effect on the 45Ca2+ influx induced by ATP. Staurosporine, an inhibitor for PKC, significantly enhanced the ATP-induced 45Ca2+ influx. Pertussis toxin inhibited the ATP-induced formation of IP3 in a dose-dependent manner in the range between 0.1 ng/ml and 1 microgram/ml. On the other hand, pertussis toxin significantly enhanced the ATP-induced 45Ca2+ influx. These results strongly suggest that extracellular ATP-induced Ca2+ influx is autoregulated due to the activation of PKC resulting from pertussis toxin-sensitive GTP-binding protein-coupled phosphoinositide hydrolysis in PC12 pheochromocytoma cells.

Adenosine Triphosphate↗
PubMed1993

Isolation and identification of anti-tumor-promoting principles from the fresh-water cyanobacterium Phormidium tenue.

Bioassay-directed fractionation of the extract of the cyanobacterium P. tenue led to the isolation of the three classes of glycolipids, viz., monogalactosyl diacylglycerol (MGDG), digalactosyl diacylglycerol (DGDG), and sulfoquinovosyl diacylglycerol (SQDG) as anti-tumor-promoters. In comparing the anti-tumor-promoting effect of the three classes of glycolipids with various acyl residues, MGDG and DGDG inhibited tumor promotion more intensely than SQDG. With respect to MGDG and DGDG, the inhibitory activity of each glycolipid possessing a single pair of acyl residues was also investigated after HPLC separation. Among the 17 tested MGDGs (1a--h) and DGDGs (2a--i), three of the DGDGs (2a, b, e) showed more potent inhibitory activity toward tumor promotion than the others.

Anticarcinogenic Agents↗
PubMed1992

Amiloride at pH 7.0 inhibits the Na(+)-driven flagellar motors of Vibrio alginolyticus but allows cell growth.

Amiloride, a specific inhibitor for the Na(+)-driven flagellar motors of alkalophilic Bacillus, is known to inhibit secondarily the growth of alkalophiles. The motility of a marine Vibrio, V. alginolyticus, was almost completely inhibited by 2 mM amiloride either at pH 7.0 or 8.5. We found that this concentration of amiloride inhibited the cell growth completely at pH 8.5 but only slightly at pH 7.0. Kinetic analysis of the inhibition of motility by amiloride at pH 7.0 showed that the inhibition was competitive with Na+ in the medium. Thus, amiloride at pH 7.0 is really a specific and useful tool for the analysis of the Na(+)-driven flagellar motors of Vibrio.

Amiloride↗
PubMed1992

Inhibition by squalene of the tumor-promoting activity of 12-O-tetradecanoylphorbol-13-acetate in mouse-skin carcinogenesis.

Squalene inhibited the effect of tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA), such as increased 32Pi incorporation into phospholipids of HeLa cell membrane, induction of Epstein-Barr-virus early antigen in Raji cell and induction of ornithine decarboxylase in mouse skin. Squalene also markedly suppressed the promoting activity of TPA on skin carcinogenesis in 7,12-dimethylbenz[a]anthracene-initiated mice.

Animals↗
PubMed1992

Potent preventive action of alpha-carotene against carcinogenesis: spontaneous liver carcinogenesis and promoting stage of lung and skin carcinogenesis in mice are suppressed more effectively by alpha-carotene than by beta-carotene.

Although beta-carotene has been considered to be a key cancer preventive agent in green and yellow vegetables, other types of carotenoids, such as alpha-carotene, may also contribute to anticarcinogenic action, since these carotenoids usually coexist with beta-carotene and are detectable in human blood and tissues. In this study, we compared the inhibitory effect of natural alpha-carotene, obtained from palm oil, with that of beta-carotene on spontaneous liver carcinogenesis in C3H/He male mice. The mean number of hepatomas per mouse was significantly decreased by alpha-carotene supplementation (per os administration in drinking water at a concentration of 0.05%, ad libitum) as compared with that in the control group (P < 0.001, Student's t test). On the other hand, beta-carotene, at the same dose as alpha-carotene, did not show any such significant difference from the control group. Furthermore, we also compared the antitumor-promoting activity of alpha-carotene with that of beta-carotene against two-stage mouse lung carcinogenesis (initiator, 4-nitroquinoline 1-oxide; promoter, glycerol). alpha-Carotene, but not beta-carotene, reduced the number of lung tumors per mouse to about 30% of that in the control group (P < 0.001, Student's t test). The higher potency of the antitumor-promoting action of alpha-carotene compared to beta-carotene was confirmed in other experimental systems; e.g., alpha-carotene was also found to have a stronger effect than beta-carotene in suppressing the promoting activity of 12-O-tetradecanoylphorbol-13-acetate on skin carcinogenesis in 7,12-dimethylbenz[a]anthracene-initiated mice. These results suggest that not only beta-carotene, but also other types of carotenoids, such as alpha-carotene, may play an important role in cancer prevention.

4-Nitroquinoline-1-oxide↗
PubMed1992

Cleavage of hexokinase II to two domains by trypsin without significant change in catalytic activity.

Hexokinase II prepared from Ehrlich-Lettre hyperdiploid tumor cells (ELD cells) was subjected to a limited digestion by trypsin. After 60 min digestion, hexokinase II (100 kDa) was completely cleaved to two fragments with the molecular weight of about 60 kDa and 40 kDa as manifested in SDS-PAGE. It was noteworthy that the enzyme activity was observed even at the time when the native enzyme molecule was no more detectable. These fragments were separated by SDS-PAGE irrespective of the presence of a reducing agent, but neither by native PAGE nor by cellulose acetate membrane electrophoresis under the nondenaturing conditions. Neither kinetic parameters such as Km values for ATP and glucose nor an ability of binding to mitochondria were changed significantly by the tryptic digestion. These results indicate that an essential conformation of hexokinase II can be restored by the self-association of two fragments produced as a result of the cleavage by trypsin at the middle of the molecule. Affinity labeling with 2',3'-dialdehyde ATP followed by the trypsin digestion showed that ATP binding site resided in the 40 kDa fragment. Furthermore, the mode of the response in the incorporation of this ATP analog to hexose phosphate, moreover, was similar to that in the catalytic activity.

Animals↗
PubMed1992

Inhibitory effects of dihydroagarofuran sesquiterpenes on Epstein-Barr virus activation.

To search for possible antitumor promoters, we carried out a primary screening of thirty-seven dihydroagarofuran sesquiterpenes from Tripterygium wilfordii Hook fil. var. regelii Makino and Euonymus sieboldianus Blume, using their possible inhibitory effects on the Epstein-Barr virus early antigen (EBV-EA) activation which is induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) in Raji cells. Some of these sesquiterpenes, triptofordin F-2 (Takaishi et al., 1988), 1,2,6,8,15-pentaacetoxy-9-benzoyloxy-4-hydroxy-beta-dihydroagarofuran and triptogelin A-1 (Takaishi et al., 1990) were observed to significantly inhibit the EBV-EA activation at low doses. Based on the results, the structural requirements for the activity of these compounds were discussed [corrected].

Anticarcinogenic Agents↗
PubMed1992

Barbiturates suppress glucose utilization by inhibition of hexokinase in neuroblastoma cells.

Exposure to the pentobarbital potently inhibited the 2-deoxy glucose uptake in cultured neuroblastoma cells. The inhibition was assumed to be due to saturation of the uptake in the early stage where the incorporation was linear in the nontreated cells. On the contrary, the incorporation of 3-O-methyl glucose, another glucose analog which is not phosphorylated by hexokinase, was not altered by the treatment with pentobarbital. These results suggest that the suppression of hexokinase is involved in the above-mentioned effect of pentobarbital.

3-O-Methylglucose↗
PubMed1992

The carboxyl-terminal region of SecE interacts with SecY and is functional in the reconstitution of protein translocation activity in Escherichia coli.

Genes encoding the C- and N-terminal regions of SecE were constructed and placed under the control of the tac promoter on plasmids. The C-terminal region of SecE (SecE-C) was sufficient for suppression of the secEcs phenotype, confirming the results of Schatz et al. (Schatz, P. J., Bieker, K. L., Ottemann, K. M., Silhavy, T. J., and Beckwith, J. (1991) EMBO J. 10, 1749-1757). SecE-C allowed the overproduction of SecY, and its overproduction was achieved when the tac-secY gene, on a plasmid, was induced, indicating that the C-terminal region is the site of interaction of SecE with SecY and that the interaction makes the two Sec proteins stable. SecE-C was purified and used with SecY for the reconstitution of protein translocation activity. SecE-C was active in the functional reconstitution. The SecE-C/SecY-dependent protein translocation absolutely required SecA and ATP as the native translocation reaction did. Quantitative analysis revealed that SecE-C was 50% as active as intact SecE. The N-terminal region of SecE (SecE-N) also suppressed in vivo the defect caused by the secEcs mutation. SecE-N was, however, inactive in the overproduction of SecY. A possible oligomeric structure of SecE is discussed.

Adenosine Triphosphate↗