Different effects of St John's wort on the pharmacokinetics of simvastatin and pravastatin.

OBJECTIVE: St John's Wort, a widely used herbal product, is an inducer of CYP3A4 and it decreases blood concentrations of CYP3A4 substrates. The effects of St John's Wort on the pharmacokinetics of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors simvastatin (an inactive lactone pro-drug) and pravastatin were determined in this study. METHODS: Sixteen healthy male subjects (n = 8 in group 1 and n = 8 in group 2) took a St John's Wort caplet (300 mg) or matching placebo three times a day for 14 days in a double-blind, crossover study. On day 14, a single oral dose of 10 mg simvastatin and 20 mg pravastatin was given to subjects in group 1 and group 2, respectively. Blood samples were obtained during a 24-hour period after the administration of each drug. RESULTS: Repeated St John's Wort treatment tended to lower plasma simvastatin concentration and significantly (P <.05) lowered concentrations of simvastatin hydroxy acid, its active metabolite. The peak concentration in plasma (ratio, 0.72 of placebo) of simvastatin hydroxy acid tended to be decreased and its area under the plasma concentration-time curve between time zero and 24 hours after administration (ratio, 0.48 of placebo) was significantly decreased (P <.05) by St John's Wort. On the other hand, St John's Wort did not influence plasma pravastatin concentration. No significant differences were observed in the elimination half-life of simvastatin or pravastatin between the placebo and St John's Wort trials. CONCLUSIONS: This study showed that St John's Wort decreases plasma concentrations of simvastatin but not of pravastatin. Because simvastatin is extensively metabolized by CYP3A4 in the intestinal wall and liver, which are induced by St John's Wort, it is likely that this interaction is partly caused by the enhancement of the CYP3A4-mediated first-pass metabolism of simvastatin in the small intestine and liver.

Effects of repeated stress on expression of interleukin-6 (IL-6) and IL-6 receptor mRNAs in rat hypothalamus and midbrain.

We examined the effects of single and repeated stress on the expression of interleukin-6 (IL-6) and IL-6 receptor (IL-6R) mRNAs in the rat midbrain and hypothalamus using reverse transcriptase-polymerase chain reaction (RT-PCR). Following a single episode of restraint stress for 4 hours (1R) or 4 hours per day on two (2R) or three (3R) consecutive days, the hypothalamus and midbrain were removed immediately and the levels of IL-6 and IL-6R mRNAs in both regions were determined. Regional differences in stress-related changes in mRNA levels were noted. The expression of IL-6 mRNA in the hypothalamus did not change in 1R group but decreased in 2R and 3R groups. The expression of IL-6R mRNA in the same region significantly diminished in all groups. In the midbrain, the expression of IL-6 mRNA increased in 1R group and decreased in 2R and 3R, while the expression of IL-6R mRNA significantly diminished in 1R and 3R groups but was not different from control in 2R group. Our findings indicate that repeated stress in rats produce changes in IL-6 and IL-6R mRNAs in the midbrain and hypothalamus that are different than those of a single stress episode.

Chronotherapy of high-dose 1,25-dihydroxyvitamin D3 in hemodialysis patients with secondary hyperparathyroidism: a single-dose study.

BACKGROUND: A high-dose oral intermittent vitamin D (pulse therapy) is widely used for the treatment of secondary hyperparathyroidism associated with kidney failure. However, hypercalcemia by vitamin D sometimes interrupts this treatment. Because serum calcium concentration possesses a circadian rhythm, a chronopharmacologic approach of vitamin D may have merit for avoidance of adverse reactions. METHODS: Six female secondary hyperparathyroidism patients receiving maintenance hemodialysis received a single oral dose of 2 microg 1,25-dihydroxyvitamin D3 at either 8 AM or 8 PM in a crossover design. Serum concentrations of ionized and total calcium, phosphate, and vitamin D3 were determined for a 48-hour period after administration. We also measured serum intact parathyroid hormone before and 48 hours after dosing as an index for efficacy. RESULTS: A single oral administration of the drug caused an increase in concentration of ionized calcium, serum calcium, and phosphate. However, the area under concentration-time curve from zero to 48 hours [AUC(0-48)] and peak concentration of these variables were markedly lower after dosing at 8 PM. Pre-dose concentrations of these variables were lower at night. The AUC(0-48) of serum vitamin D3 of the morning and night trials did not differ significantly. Reduction of intact parathyroid hormone concentration was also similar between the two trials. CONCLUSION: The administration of vitamin D3 at night may reduce the occurrence of hypercalcemia and hyperphosphatemia in patients with secondary hyperparathyroidism, whereas the pharmacokinetics and intact parathyroid hormone-lowering effect of the drug does not vary with dosing time.

Effects of restraint stress on alpha(1) adrenoceptor mRNA expression in the hypothalamus and midbrain of the rat.

We examined the effects of restraint stress on alpha(1) adrenoceptor mRNA expression in the rat brain using reverse transcriptase-polymerase chain reaction (RT-PCR). After rats had been restrained for 10, 30, 60, 120 or 240 min, the hypothalamus and midbrain were removed immediately and alpha(1) adrenoceptor mRNA levels in these regions were determined by RT-PCR. Blood samples were also collected for simultaneous measurement of serum adrenocorticotropic hormone (ACTH) and corticosterone. Restraint stress resulted in a variety of changes in the hypothalamus and midbrain. In the hypothalamus, 30 and 60 min of stress resulted in a significant fall in the level of alpha(1) adrenoceptor mRNA relative to the control. This was associated with a rise in serum ACTH and corticosterone. In the midbrain, significant elevation of alpha(1) adrenoceptor mRNA was noted after 60, 120 and 240 min of restraint stress. Our findings indicated that the influence of restraint stress on alpha(1) adrenoceptor mRNA level in the hypothalamus is different to that of the midbrain region in rats.

Effects of single and repeated stresses on the expression of mRNA for alpha1-adrenoceptors in the rat hypothalamus and midbrain.

We examined the effects of single or repeated stress on the expression of mRNA for alpha1-adrenoceptors in the rat hypothalamus and midbrain using the reverse transcriptase-polymerase chain reaction (RT-PCR). Single stress significantly increased the mRNA level for alpha1-adrenoceptors in the midbrain, but had no effect on mRNA levels in the hypothalamus. Repeated stress significantly decreased mRNA levels for alpha1-adrenoceptors in both regions.

Effects of a long-term inhalation of fragrances on the stress-induced immunosuppression in mice.

The aim of this study was to determine the effects of the long-term application of various fragrances on the suppression of immune response induced by high-pressure stress in mice. The immune response was analyzed based on plaque-forming cell (PFC) count, using mice sensitized with sheep red blood cells. The decreased PFC involving thymic involution induced by high-pressure stress in mice was restored by exposing the stressed mice to tuberose, lemon, oakmoss and labdanum for 24 h following exposure to stress. The decreased PFC and thymic involution from stress were restored by exposure to lemon and oakmoss, but not to tuberose and labdanum when the mice were exposed to those fragrances continuously for 3 weeks before the stress was given, followed by exposure to the same fragrances for 24 h after the stress. The decreased PFC and thymic involution from stress were restored by exposure to lemon and labdanum for 24 h after the stress, but not to tuberose over 3 weeks before the stress was given. These data suggest that the neuroimmunomodulatory effects of fragrances may be affected by tolerance depending on the kinds of fragrances in the case of a long-term application.

The expressions of mRNAs for interleukin-6 (IL-6) and the IL-6 receptor (IL-6R) in the rat hypothalamus and midbrain during restraint stress.

Over the past few years, it has been reported that physical and psychological stress elevate plasma interleukin-6 (IL-6), and that neural cells can produce IL-6 and have receptors for IL-6 (IL-6R). However, it is unknown whether IL-6 plays a role in regulating the functions of neural cells in response to stress. We demonstrated recently, using the reverse transcriptase-polymerase chain reaction (RT-PCR), that the levels of mRNAs for IL-6 and IL-6R in the rat brain are changed by restraint stress for four hours. In the present study, we investigated the expression of mRNAs for IL-6 and the IL-6R in the rat hypothalamus and midbrain during restraint stress. After rats had been restrained for 10, 30, 60, 120 or 240 min, the hypothalamus and midbrain were removed immediately and levels of IL-6 mRNA and of IL-6R mRNA in these regions were determined by RT-PCR. The expression of mRNAs for IL-6 and IL-6R in both regions was reduced after short-term (30-60 min) restraint stress and tended to return toward the control level after 120 min restraint stress. After long-term (240 min) restraint stress, the level of IL-6 mRNA was significantly increased in the midbrain, while the level of IL-6R mRNA was significantly reduced in both regions. These findings suggest that the need for IL-6 might decline after short-term restraint stress and, moreover, that the synthesis and secretion of IL-6 might be enhanced and IL-6 might be needed as a neurotrophic factor in the midbrain after long-term stress.

The influence of restraint stress on the expression of mRNAs for IL-6 and the IL-6 receptor in the hypothalamus and midbrain of the rat.

Using the reverse transcriptase-polymerase chain reaction (RT-PCR), we investigated the influence of restraint stress on the expression of the mRNA for interleukin-6 (IL-6) and the mRNA for the IL-6 receptor (IL-6R) in the rat brain. After rats had been restrained for 4 hours, the hypothalamus and midbrain were removed at fixed intervals up to 24 hours, and levels of IL-6 mRNA and of IL-6R mRNA in these regions were determined by RT-PCR. Restraint stress significantly enhanced the expression of IL-6 mRNA and reduced that of IL-6R mRNA in the midbrain, whereas the stress caused the reduced expression of IL-6R mRNA without any change in the level of IL-6 mRNA in the hypothalamus. After the stress, the expression of mRNAs for IL-6 and IL-6R continued to diminish in both regions. These findings indicate that the levels of mRNAs for both of IL-6 and IL-6R in the rat brain can be influenced by restraint stress.

Hepatotoxicity of gardenia yellow color in rats.

Acute toxicity by gardenia yellow color was studied in rats. Oral administration of the colorant at doses of 800 mg/kg up to 5000 mg/kg caused diarrhea and increases in serum alanine aminotransferase and aspartate aminotransferase activities in a dose-dependent manner. After 24 h of oral treatment with 2000 mg/kg of the colorant, liver showed partially hemorrhagic changes and the intestinal tract, especially the duodenum, appeared blue. The toxicity induced by the colorant was stronger by oral administration than by intraperitoneal administration. The content of geniposide, an iridoid compound, was estimated to be 28% of the colorant, and this iridoid accounted for almost all the hepatotoxic activity of the colorant.

Isolation and identification of 1 alpha- and 23-hydroxylated metabolites of 25-hydroxy-24-oxovitamin D3 from in vitro incubates of chick kidney homogenates.

Five major metabolites (peaks I-V) of 25-hydroxy-24-oxovitamin D3 (25(OH)24-oxo-D3) have been isolated in pure form from in vitro incubates containing kidney homogenates of vitamin D-deficient chicks and chicks given 65 nmol of vitamin D3; peaks II, III, and V are from vitamin D-deficient chicks and peaks I, II, and IV are from vitamin D-supplemented birds. The structures of the metabolites were unequivocally identified as 23,25-dihydroxy-24-oxo-vitamin D3 (peak I), 24,25-dihydroxyvitamin D3 (24,25(OH)2D3) (peak II), 1 alpha, 25-dihydroxy-24-oxovitamin D3 (peak III), 23,24,25-trihydroxyvitamin D3 (peak IV), and 1 alpha,24,25-trihydroxyvitamin D3 (peak V) by means of ultraviolet absorption spectrometry, mass spectrometry, and specific chemical reactions. It is concluded that 25(OH)24-oxo-D3 is further hydroxylated at the 1 alpha-position in the kidney of vitamin D-deficient chicks and at the 23-position in that of vitamin D-supplemented animals. Formation of 24,25(OH)2D3 from 25(OH)24-oxo-D3 in both vitamin D-deficient and -supplemented animals provides evidence for the presence of an enzyme to reduce the 24-oxo group irrespective of the vitamin D status.

Chemical synthesis, biological activity, and metabolism of 25-hydroxy-24-oxovitamin D3.

25-Hydroxy-24-oxovitamin D3 (25(OH)24-oxo-D3), a metabolite of 25-hydroxyvitamin D3, has been chemically synthesized. The ultraviolet, mass, infrared, and proton nuclear magnetic resonance spectra of the 25(OH)24-oxo-D3 were identical with those of the natural product isolated from chick kidney incubates. The oxo compound showed biological activity similar to 24,25-dihydroxyvitamin D3 (24,25(OH)2D3) in vitamin D-deficient chicks in enhancing intestinal calcium transport and bone calcium mobilization activities. Although 25(OH)24-oxo-D3 partially restored the impaired eggshell weights of Japanese quails fed a vitamin D-deficient diet, it was much less potent than 25-hydroxyvitamin D3 or 1 alpha, 25-dihydroxyvitamin D3. In addition, there was no effect on the calcification of medullary bone. When 25(OH)24-oxo[3H]D3 was incubated with kidney homogenates from vitamin D-deficient chicks, it was metabolized to [3H]1 alpha, 24,25-trihydroxyvitamin D3 and a metabolite which was eluted in a region between authentic 24R,25(OH)2D3 and 1 alpha, 25-dihydroxyvitamin D3 on high pressure liquid chromatography. In the incubates of kidney homogenates from vitamin D-supplemented chicks, those metabolites were not detected. In vitamin D-supplemented chicks, the recovery of radioactivity in the chloroform phase extracted by the method of Bligh and Dyer was only 50%, while that in vitamin D-deficient chicks was 87%. Moreover, the radioactivity eluted in the 25(OH)24-oxo-D3 fraction from vitamin D-supplemented chicks was only one-fifth of that from vitamin D-deficient birds. The present results indicate that the 24-oxidation of 24,25(OH)2D3 may be a route of inactivation of vitamin D3.

An examination of the importance of 24-hydroxylation to the function of vitamin D during early development.

To investigate the biological importance of 24R-hydroxylation of 25-hydroxyvitamin D to the early development of rats, the potency of 24,24-difluoro-25-hydroxyvitamin D3 had been compared to that of 25-hydroxyvitamin D3 in young rat pups born to vitamin D-deficient mothers. 24,24-Difluoro-25-hydroxyvitamin D3 and 25-hydroxyvitamin D3 were equally active in stimulating active calcium transport in the intestine, maintaining normal concentrations of calcium and phosphorus in the plasma and promoting bone growth and mineralization. These results provide strong evidence that the presence of a hydroxyl group at the 24 position of vitamin D3 is not required for the maintenance of calcium-phosphate homeostasis during growth and in the development and mineralization of bone.