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991.
Antioxidative Activity of 5,6,7,8-Tetrahydrobiopterin and its Inhibitory Effect on Paraquat-Induced Cell Toxicity in Cultured Rat Hepatocytes 总被引:3,自引:0,他引:3
The in vitro antioxidative activity of 5,6,7,8-tetrahydrobiopterin (BPH4) was measured and the ability of BPH4 to prevent paraquat-induced cell damage was examined in cultured hepatocytes. The scavenging activity of BPH4 against superoxide anion radicals was assayed in two systems, i.e., xanthine/xanthine oxidase (X/XOD) and rat macrophage/phorbol myristate acetate (MξPMA) radical-generating systems. BPH4 showed an extremely strong superoxide anion radical-scavenging activity in both assay systems. Biopterin (BP) itself did not show any activity in the X/XOD system, but was effective in the MξPMA system. The antioxidative activities of BPH4 against both superoxide anion and hydroxyl radicals were confirmed by spin trapping-ESR spectrometry. BPH4 also protected rat brain homogenate against auto-oxidation. We further examined the effect of BPH4 on paraquat-induced cell toxicity in cultured rat hepatocytes. The paraquat-induced elevation of the release of lactate dehydrogenase (LDH), a marker enzyme for cytotoxicity from cultured hepatocytes was suppressed by BPH4 in a dose-dependent manner. The elevation of lipid peroxides simultaneously induced by paraquat was also inhibited by BPH4 in the same manner. These results suggest that BPH4 might be useful in the treatment of various diseases whose pathogenesis is active oxygen-related. 相似文献
992.
M. A. Lelu K. Klimaszewska G. Pflaum C. Bastien 《In vitro cellular & developmental biology. Plant》1995,31(1):15-20
Summary Cotyledonary somatic embryos ofLarix × leptoeuropaea that developed after various maturation times on media containing abscisic acid showed different frequencies of conversion
into plants. Drying of these somatic embryos under high relative humidity (RH) before germination improved plantlet recovery
and eliminated differences in the performance of somatic embryos matured for different times. However, dehydration of somatic
embryos under 98% RH to a water content below that of zygotic embryos excised from mature seeds (0.97 and 1.36 g H2O/g dry weight, respectively) showed a strong positive correlation between longer maturation time and desiccation tolerance.
Drying somatic embryos at 4° C under 59% RH for 1 wk resulted in desiccation to a water content of 0.30 g H2O/g dry weight, which was the closest to the hydration state of zygotic embryos in dried, stored seeds (0.20 g H2O/g dry weight). Under this condition, only somatic embryos matured for 5 wk germinated and produced plantlets at a relatively
high frequency (73 and 41%, respectively). 相似文献
993.
D. M. Stresser G. S. Bailey D. E. Williams L. F. Bjeldanes 《Journal of biochemical and molecular toxicology》1995,10(4):191-201
Dietary indole-3-carbinol inhibits carcinogenesis in rodents and trout. Several mechanisms of inhibition may exist. We reported previously that 3,3′-diindolylmethane, an in vivo derivative of indole-3-carbinol, is a potent noncompetitive inhibitor of trout cytochrome P450 (CYP) 1A-dependent ethoxyresorufin O-deethylase with Ki values in the low micromolar range. We now report a similar potent inhibition by 3,3′-diindolylmethane of rat and human CYP1A1, human CYP1A2, and rat CYP2B1 using various CYP-specific or preferential activity assays. 3,3′-Diindolylmethane also inhibited in vitro CYP-mediated metabolism of the ubiquitous food contaminant and potent hepatocarcinogen, aflatoxin B1. There was no inhibition of cytochrome c reductase. In addition, we found 3,3′-diindolylmethane to be a substrate for rat hepatic microsomal monooxygenase(s) and tentatively identified a monohydroxylated metabolite. These observations indicate that 3,3′-diindolylmethane can inhibit the catalytic activities of a range of CYP isoforms from lower and higher vertebrates in vitro. This broadly based inhibition of CYP-mediated activation of procarcinogens may be an indole-3-carbinol anticarcinogenic mechanism applicable to all species, including humans. © 1995 John Wiley & Sons, Inc. 相似文献
994.
E. Fabbri A. Capuzzo A. Gambarotta T.W. Moon 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》1995,112(4):643-651
Epinephrine (EPI) is thought to act by stimulating adenylyl cyclase (ACase) and cAMP production through β-adrenoceptors in the liver of more primitive vertebrates. Recent observations, however, point to an involvement of α1-adrenoceptors in EPI action, at least in some fish species. The role of the α1- and β-adrenergic transduction pathways has been investigated in rainbow trout (Oncorhynchus mykiss) hepatic tissue. Radioligand-binding assays with the β-adrenergic antagonist 3H-CGP-12177 using hepatic membranes purified on a discontinuous sucrose gradient confirmed the presence of β-adrenoceptors (Kd0.36 nM, Bmax 8.61 fmol · mg−1 protein). We provide the first demonstration of α1-adrenoceptors in these same membranes; analysis of binding data with the α1-adrenergic antagonist 3H-prazosin demonstrated a single class of binding sites with a Kdof 15.4 nM and a Bmax of 75.2 fmol · mg−1 protein. There is a straight correlation between β-adrenoceptor occupancy, ACase activation and cAMP production. On the contrary, the role of inositol 1,4,5-trisphosphate (IP3) has to be elucidated; in fact, despite the presence of specific microsomal binding sites for IP3 (Kd 6.03 nM, Bmax 90.2 fmol · mg−1 protein), its cytosolic concentration was not modulated by EPI. On the other hand, we have previously shown in American eel and bullhead hepatocytes that α1-adrenergic agonists are able to increase intracellular concentrations of IP3 and Ca2+ and to activate glycogenolysis. These data suggest a marked variation in the liver of different fish both in terms of α1-binding sites affinity and of α1-adrenoceptor/IP3/Ca2+ transduction systems. 相似文献
995.
Stuart L. Myers Richard Turnage Kevin Kadesky Lori Bartula Angela Riva Barbara Kalley-Taylor 《Prostaglandins & other lipid mediators》1995,50(1)
This study examines the hypothesis that PAF stimulates release of PGI2 from inflamed rabbit gallbladder explant cell cultures. New Zealand white rabbits underwent bile duct ligation for 72 h (72 h BDL), or sham operation, Sham and 72 h BDL gallbladder explants were placed in culture, and the cells grown to 75% confluence. The cells were exposed to increasing concentrations of PAF for 60 min. The media analyzed for eicosanoid release by EIA and the cells analyzed for cyclooxygenase and prostacyclin synthase content by immunoblot analysis. PAF increased release of 6-keto-PGF1α from the 72 h BDL gallbladder cell cultures in a dose-related manner which was inhibited by indomethacin preincubation by 90%. The increased 72 h BDL cell release of 6-keto-PGF1α was not associated with changes in the content of cyclooxygenase or prostacyclin synthase. PAF did not alter eicosanoid release from sham control cell cultures. These data suggest that PAF can only up-regulate endogenous 6-keto-PGF1α release from the 72 h BDL cells that had been previously stimulated by inflammation. PAF may thus contribute to gallbladder distention and injury by chronic stimulation of inflamed gallbladder PGI2 release. 相似文献
996.
A chlorine-containing non-protein amino acid which was recently discovered from the fruit bodies ofAmanita gymnopus (2S)-2-amino-5-chloro-4-hydroxy-5-hexenoic acid, was isolated and crystallized for the first time from the fruit bodies of an unknown member ofAmanita belonging to the sectionRoanokenses, subsectionSolitariae. The results of elementary analyses, determination of optical rotations,1H- and13C-NMR-spectra, and some chemical reactions supported an earlier proposed structure.Part 24 in the series Biochemical studies of nitrogen compounds in fungi. for Part 23, see Hatanaka, S. I. et al. 1994. this journal35: 391–394. 相似文献
997.
Plant regeneration from protoplasts of Lilium × formolongi cv. Azusa was achieved by utilizing suspension cultures of meristematic nodular cell clumps with a high plant regeneration ability. Creamy-white calli with embryogenic potential were initially induced from the seeds on Murashige and Skoog (MS) medium containing 4.1 μM picloram. The calli were then transferred into a liquid medium of the same composition, in which they turned into compact cell clumps which consisted of meristematic nodules. Protoplasts were readily isolated from these meristematic nodular cell clumps. Colonies were successfully formed from the protoplasts by embedding in 0.1% gellan gum-solidified MS medium containing 4.1 μM picloram and 0.5 M glucose. They regenerated shoots and roots on MS medium containing 2.2 μM benzylaminopurine (BAP). The plants thus obtained produced flowers with normal fertile pollen 8 months after successful transfer into soil. These plants had normal chromosome numbers (2n = 24) but had shorter leaves than original plants. They set seeds after as well as cross pollination. 相似文献
998.
The temperature dependence of high voltage activated Ca2+ channels has been investigated in cultured dorsal root ganglion neurones from chick embryos, using the cell-attached patch-clamp technique. The dihydropyridine sensitive L-type Ca2+ channel had a conductance of 23 pS, with 110 mM Ba2+ as charge carrier and in the presence of 3 M Bay K 8644. When the temperature was raised from 15 to 30 °C, the unitary channel current amplitude increased, with Q10 value equal to 1.4. The rising phase of the averaged single-channel current became faster, with Q10 value 2.7, whereas the decay phase showed a lower temperature sensitivity. Channel open probability decreased according to an exponential distribution of open and closed times. A second type of Ca2+ channel was identified, which was DHP-insensitive and had a lower conductance with a mean value equal to 13 pS. For the current amplitude, the Q10 value was 1.3. Both activation and inactivation kinetics were strongly accelerated by an increase in temperature. The corresponding time constants gave Q10 values equal to 5.9 for activation, and 2.0 for inactivation. Peak channel open probability was highly sensitive to a change in temperature, with a Q10 value of 1.6. Finally, in -conotoxin GVIA pre-treated neurones, a non-inactivating DHP-insensitive Ca2+ channel with the lowest unitary conductance (10 pS) and a much lower temperature dependence was recorded. Single-channel current was increased by heating, with Q10 value 1.3, whereas the channel kinetics were almost unaffected by temperature. Our data are consistent with the assumption that the different temperature dependence of the Ca2+ channel behaviours may be explained by separate gating processes of three types of Ca2+ channels. 相似文献
999.
Summary 1. We examined the actions of mercury (Hg2+) and zinc (Zn2+) on voltage-activated calcium channel currents of cultured rat dorsal root ganglion (DRG) neurons, using the whole-cell patch clamp technique.2. Micromolar concentrations of both cations reduced voltage-activated calcium channel currents. Calcium channel currents elicited by voltage jumps from a holding potential of –80 to 0 mV (mainly L- and N-currents) were reduced by Hg2+ and Zn2+. The threshold concentration for Hg2+ effects was 0.1 µM and that for Zn2+ was 10µM. Voltage-activated calcium channel currents were abolished (>80%) with 5µM Hg2+ or 200µM Zn2+. The peak calcium current was reduced to 50% (IC50) by 1.1µM Hg2+ or 69µM Zn2+. While Zn2+ was much more effective in reducing the T-type calcium channel current—activated by jumping from –80 to –35 mV—Hg2+ showed some increased effectiveness in reducing this current.3. The effects of both cations occurred rapidly and a steady state was reached within 1–3 min. While the action of Zn2+ was not dependent on an open channel state, Hg2+ effects depended partially on channel activation.4. While both metal cations reduced the calcium channel currents over the whole voltage range, some charge screening effects were detected with Hg2+ and with higher concentrations (>100µM) of Zn2+.5. As Zn2+ in the concentration range used had no influence on resting membrane currents, Hg2+ caused a clear inward current at concentrations µM.6. In the present study we discuss whether the actions of both metals on voltage-activated calcium channel currents are mediated through the same binding site and how they may be related to their neurotoxic effects. 相似文献
1000.
A simple procedure is described for the elimination ofO-linked glycans from bovine submaxillary mucin under non-reducing conditions, using triethylamine in aqueous hydrazine. The glycans were isolated as the hydrazones, which were converted to the reducing glycans by exchange with acetone in neutral aqueous solution. The glycan alditols obtained after reduction corresponded to those obtained by the reductive -elimination ofO-glycans. 相似文献