Biomedical subjects

H W Chen

Publications and source records attributed to H W Chen.

At least 73 records · Page 4Linked to original sources

PubMed1998

In vitro maturation of follicular oocytes of the Giant Panda (Ailuropoda melanoleuca): a case report.

The Giant Panda is an endangered species that would benefit from biotechnological assistance in reproduction. However, because there are only a few of these animals left in the world, scientists hesitate to use them for research procedures. We were fortunate to obtain ovaries from a Giant Panda that died of hepatic cirrhosis during the nonbreeding season. Oocytes were harvested within 4 h of death by dissecting the ovarian cortex in physiological saline and collecting the cumulus-oocyte complexes from the fluid, and then were classified into large (> 125 microns) and small (100 to 124 microns) follicular oocytes and placed in TCM199 supplemented with FSH (10 micrograms/mL) and LH (20 micrograms/mL). After culture for 22 h at 37 degrees C in air with 5% CO2, response was evaluated by growth of oocytes and presence of the first polar body. Of the 26 large follicular oocytes that were harvested, 12 were considered suitable for IVM, and 14 were degenerated, had a broken zona pellucida or had lost some cytoplasm. Of the 12 cultured oocytes, all grew to a mean diameter of 141.1(SD = +/- 6.7, n = 12), and 4 released the first polar body. None of the small follicular oocytes showed growth or other signs of maturation. We conclude from our preliminary results that it is possible to obtain functional Giant Panda oocytes from ovaries obtained post mortem during the nonbreeding season.

Animals↗

PubMed1998

Effect of dietary lipid on gamma-glutamyl transferase-positive foci during hepatocarcinogenesis in rats.

The effect of dietary lipid on gamma-glutamyl transferase-positive (GGT-positive) foci was investigated. Female Sprague-Dawley rats were dosed with diethylnitrosamine (15 mg/kg) at 24 h of age. After weaning, they were fed nutritionally complete semipurified diets for 3 months. Rats fed 15% corn oil had significantly lower hepatic phospholipid eicosapentaenoate and docosahexaenoate than rats fed 7.5% corn oil plus 7.5% fish oil, 5% corn oil plus 10% fish oil (P < 0.05). However, rats fed 15% corn oil had significantly greater hepatic phospholipid arachidonate than rats fed the other two diets (P < 0.05), suggesting that n-3 polyunsaturated fatty acids were incorporated into hepatic phospholipid at the expense of n-6 polyunsaturated fatty acids. Hepatic PGF2alpha content was significantly greater in rats fed 15% corn oil than in rats fed the other two diets (P < 0.05). Rats fed fish oil had significantly lower hepatic vitamin E content than rats fed corn oil (P < 0.05). Hepatic lipid peroxidation (TBARS) tended to increase with increased dietary fish oil (P < 0.05). Dietary lipid did not influence GGT-positive foci area or number. In conclusion, dietary lipid affected hepatic PGF2alpha production, however, showed no effect on GGT-positive foci area and number. This may suggest that PGF2alpha is not the underlying mechanism for GGT-positive foci during hepatocarcinogenesis.

Animals↗

PubMed1998

The possible mechanisms of the antiproliferative effect of fullerenol, polyhydroxylated C60, on vascular smooth muscle cells.

1. The possible mechanisms of the antiproliferative effect of polyhydroxylated fullerene (fullerenol), a novel free radical trapper, were studied in rat vascular smooth muscle cells (A7r5 cells) and compared with the effect of ascorbic acid. 2. Fullerenol-1 and ascorbic acid inhibited the proliferative responses in a number of cells, including rat aortic smooth muscle cells (A7r5 cells), human coronary artery smooth muscle cells, and human CEM lymphocytes (CEM cells) in a concentration dependent manner. 3. At the concentration range of 10(-6) to 10(-2) M, fullerenol-1 and ascorbic acid concentration-dependently inhibited the proliferative responses stimulated by serum in A7r5 cells. Fullerenol-1 was more potent than ascorbic acid. 4. The production of O2- induced by alloxan, a diabetogenic compound, was reduced by fullerenol-1 (10(-4) M) in the presence of A7r5 cells. 5. The cytosolic protein kinase C activity of A7r5 cells stimulated by phorbol ester was reduced by 10(-3) M fullerenol-1, but not ascorbic acid (10(-4)-10(-2) M) and fullerenol-1 at lower concentrations (10(-6)-10(-4) M). 6. In contrast, the membraneous protein tyrosine kinase activity of A7r5 cells stimulated by foetal calf serum was significantly reduced by fullerenol-1 (10(-6)-10(-3) M) and ascorbic acid (10(-4)-10(-2) M). Again, the inhibitory activity of fullerenol-1 was greater than that of ascorbic acid. 7. Our results demonstrate that fullerenol-1 and ascorbic acid exhibit inhibitory effects on transduction signals in addition to their antioxidative property. It is suggested that the antiproliferative effect of fullerenol-1 on vascular smooth muscle cells may partly be mediated through the inhibition of protein tyrosine kinase.

Animals↗

PubMed1998

Effect of curcumin on cell cycle progression and apoptosis in vascular smooth muscle cells.

1. The possible mechanisms of the antiproliferative and apoptotic effects of curcumin (diferuloylmethane), a polyphenol in the spice turmeric, on vascular smooth muscle cells were studied in rat aortic smooth muscle cell line (A7r5). 2. The proliferative response was determined from the uptake of [3H]-thymidine. Curcumin (10(-6)-10(-4) M) inhibited serum-stimulated [3H]-thymidine incorporation of both A7r5 cells and rabbit cultured vascular smooth muscle cells in a concentration-dependent manner. Cell viability, as determined by the trypan blue dye exclusion method, was unaffected by curcumin at the concentration range 10(-6) to 10(-5) M in A7r5 cells. However, the number of viable cells after 10(-4) M curcumin treatment was less than the basal value (2 x 10(5) cells). 3. To analyse the various stages of the cell cycle, [3H]-thymidine incorporation into DNA was determined every 3 h. After stimulation with foetal calf serum, quiescent A7r5 cells started DNA synthesis in 9 to 12 h (G1/S phase), then reached a maximum at 15 to 18 h (S phase). Curcumin (10(-6)-10(-4) M) added during either the G1/S phase or S phase significantly inhibited [3H]-thymidine incorporation. 4. Following curcumin (10(-6)-10(-4) M) treatment, cell cycle analysis utilizing flow cytometry of propidium iodide stained cells revealed a G0/G1 arrest and a reduction in the percentage of cells in S phase. Curcumin at 10(-4) M also induced cell apoptosis. It is suggested that curcumin arrested cell proliferation and induced cell apoptosis, and hence reduced the [3H]-thymidine incorporation. 5. The apoptotic effect of 10(-4) M curcumin was also demonstrated by haematoxylin-eosin staining, TdT-mediated dUTP nick end labelling (TUNEL), and DNA laddering. Curcumin (10(-4) M) induced cell shrinkage, chromatin condensation, and DNA fragmentation. 6. The membranous protein tyrosine kinase activity stimulated by serum in A7r5 cells was significantly reduced by curcumin at the concentration range 10(-5) to 10(-4) M. On the other hand, the cytosolic protein kinase C activity stimulated by phorbol ester was reduced by 10(-4) M curcumin, but unaffected by lower concentrations (10(-6)-10(-5) M). 7. The levels of c-myc, p53 and bcl-2 mRNA were analysed using a reverse transcription-polymerase chain reaction (RT-PCR) technique. The level of c-myc mRNA was significantly reduced by curcumin (10(-5)-10(-4) M) treatment. And, the level of bcl-2 mRNA was significantly reduced by 10(-4) M curcumin. However, the alteration of the p53 mRNA level by curcumin (10(-5)-10(-4) M) treatment did not achieve significance. The effects of curcumin on the levels of c-myc and bcl-2 mRNA were then confirmed by Northern blotting. 8. Our results demonstrate that curcumin inhibited cell proliferation, arrested the cell cycle progression and induced cell apoptosis in vascular smooth muscle cells. Curcumin may be useful as a template for the development of drugs to prevent the pathological changes of atherosclerosis and post-angioplasty restenosis. Our results suggest that the antiproliferative effect of curcumin may partly be mediated through inhibition of protein tyrosine kinase activity and c-myc mRNA expression. And, the apoptotic effect may partly be mediated through inhibition of protein tyrosine kinase activity, protein kinase C activity, c-myc mRNA expression and bcl-2 mRNA expression.

Animals↗

PubMed1998

Effect of vitamin E on rat hepatic cytochrome P-450 activity.

Hepatic cytochrome P-450 activity has been shown to be affected by various dietary factors including vitamin E. However, reports of the effect of dietary vitamin E on cytochrome P-450 activity have been inconsistent. The aim of the present study was to investigate the influence of dietary vitamin E on rat hepatic cytochrome P-450 activity. Three groups of six male weanling Sprague-Dawley rats were fed semipurified diets containing 0, 100, or 1,500 ppm vitamin E for eight weeks. Vitamin E was given in the form of alpha-tocopheryl acetate. Dietary vitamin E significantly affected liver vitamin E content (p < 0.05) but had no effect on rat hepatic total P-450 content, N-nitrosodimethylamine demethylase, and NADPH-cytochrome-P-450 reductase activities. Hepatic pentoxyresorufin O-dealkylase and glutathione S-transferase activities were significantly greater in rats fed 100 and 1,500 ppm vitamin E than in rats fed no vitamin E (p < 0.05). Dietary vitamin E induced changes in hepatic phospholipid fatty acid composition. Hepatic phospholipid linoleate was significantly greater in rats fed 0 and 1,500 ppm vitamin E than in rats fed 100 ppm vitamin E (p < 0.05). Hepatic phospholipid eicosapentaenoate was increased significantly by dietary vitamin E (p < 0.05). Hepatic thiobarbituric acid-reactive substance was significantly greater in rats fed no vitamin E than in rats fed 100 and 1,500 ppm vitamin E (p < 0.05). The results suggest that vitamin E may influence cytochrome P-450 IIB1 enzyme activity and may affect hepatic phospholipid fatty acid composition.

Animals↗

PubMed1998

alpha-Tocopherol acetate supplementation enhances rat hepatic cytochrome PROD activity in the presence of phenobarbital induction.

Hepatic cytochrome P-450 enzymes play important roles in bioactivation of chemical carcinogens, biotransformation of many endogenous compounds, and detoxification of numerous xenobiotics. These enzyme activities have been shown to be regulated by various dietary factors. In our previous study, hepatic cytochrome pentoxyresorufin O-dealkylase (PROD) activity was decreased in rats fed an alpha-tocopherol acetate-deficient diet compared with rats fed alpha-tocopherol acetate-adequate or -supplemented diets. The objective of the present study was to investigate whether the modulatory effect of dietary alpha-tocopherol acetate on hepatic cytochrome PROD activity is influenced by the presence of phenobarbital. Weanling male Sprague-Dawley rats were fed the AIN-76 diet for four days, fasted for two days, then fed semipurified diets that were alpha-tocopherol acetate deficient, adequate, or supplemented with 5 and 15 g/kg alpha-tocopherol acetate for four days. Liver and plasma alpha-tocopherol concentrations were dose dependently regulated by dietary alpha-tocopherol acetate level. Inhibition of lipid peroxidation by dietary alpha-tocopherol acetate was dose dependent. Hepatic total cytochrome P-450 content was significantly greater in rats fed diets supplemented with 5 and 15 g/kg alpha-tocopherol acetate than in rats fed an alpha-tocopherol-adequate diet (p < 0.05). Hepatic cytochrome PROD activity was significantly greater in rats fed diets supplemented with 5 and 15 g/kg alpha-tocopherol acetate than in rats fed alpha-tocopherol acetate-deficient and -adequate diets (p < 0.05). These results suggest that, in the presence of phenobarbital, dietary alpha-tocopherol acetate efficiently affects tissue alpha-tocopherol levels and inhibits lipid peroxidation and that diets supplemented with 5 or 15 g/kg alpha-tocopherol acetate enhance hepatic cytochrome PROD activity compared with alpha-tocopherol acetate-deficient or -adequate diets.

Animals↗

PubMed1998

Effect of dietary vitamin E on antioxidant status and antioxidant enzyme activities in Sprague-Dawley rats.

The effect of dietary vitamin E on plasma, red blood cells (RBC), hepatic antioxidant status, and antioxidant enzyme activities was investigated. Three groups of six Sprague-Dawley rats were fed 0, 100, or 1,500 ppm vitamin E for eight weeks. Plasma alpha-tocopherol level was increased significantly by increasing dietary vitamin E (p < 0.05). Plasma lipid peroxidation (thiobarbituric acid-reactive substances) stimulation by 1 mM t-butyl hydroperoxide was correlated with dietary vitamin E level and was significantly greater in rats fed no vitamin E than in rats fed 100 or 1,500 ppm vitamin E (p < 0.05). RBC reduced glutathione (GSH) level was positively correlated with dietary vitamin E and was significantly greater in rats fed 1,500 ppm vitamin E than in rats fed 0 or 100 ppm vitamin E (p < 0.05). RBC oxidized glutathione was negatively correlated with dietary vitamin E. GSH redox status was expressed as the GSH-to-total GSH ratio; the ratio was also positively correlated with dietary vitamin E and was significantly greater in rats fed 1,500 ppm vitamin E than in rats fed no vitamin E (p < 0.05). For antioxidant enzymes, superoxide dismutase activity in hepatic cytosolic fraction was significantly greater in rats fed 1,500 ppm vitamin E than in rats fed 100 ppm vitamin E. Hepatic GSH reductase activity was significantly greater in rats fed 100 ppm vitamin E than in rats fed no vitamin E (p < 0.05). Dietary vitamin E had no effect on plasma vitamin C and protein thiol levels. In the systems studied, results indicated that dietary vitamin E selectively influences plasma vitamin E level, RBC GSH status, and hepatic cytosolic superoxide dismutase and GSH reductase activities.

Animals↗

PubMed1998

[Studies on late replication bands of giant panda (Ailuropoda melanoleuca) chromosomes].

Using the cultured giant panda (Ailuropoda melanoleuca) lymphocytes as experimental material, we carried out the terminal marking on the chromosomes which were in replication by adding BrdU (a final concentration: 10 micrograms/ml) about four hours before harvesting the cells. The chromosomes marked by BrdU were proceeded by staining with acridine orange solution (0.05%), irradiated by ultraviolet and counter-stained by Giemsa, we obtained clear chromosomes replication patterns. According to the different replication bands, every chromosome's characteristics in late replication behavior could be identified. In the two X chromosomes of female individual, one X chromosome is obviously much later than the other one. Especially in the large area near centromere on the long arm of late replicating X chromosome. In the male individual, there is also a large area on the long arm of chromosome Y which replicates very late, but the end of long arm of chromosome Y replicates much earlier.

Animals↗

PubMed1998

Adrenal hemangioma: two cases report.

Adrenal hemangiomas are rare, nonfunctioning benign tumors. They are well circumscribed and comprise of closely adjacent vascular channels of varying sizes that are lined with a single layer of endothelium. When they occur, they are frequently located in the skin and liver. There are no characteristic symptoms of adrenal hemangioma unless the tumor reaches a size large enough to exert pressure. To our knowledge, there have been 14 clinical cases of adrenal hemangiomas reported, all with similar pathologic features. We present 2 additional cases, which were identified incidentally after non-urologic complaints (epigastric fullness and low back pain in patient 1 and patient 2, respectively). In these 2 patients, the tumors were surgically removed and diagnosed postoperatively as adrenal hemangiomas. Preoperative radiologic findings on plain film and abdominal computerized tomography showed the characteristic round calcifications with translucent centers, typical of phleboliths, to be pathognomonic of adrenal gland hemangiomas. Unfortunately, this characteristic feature of adrenal hemangioma existed in only one case report. In conclusion, the preoperative diagnosis of adrenal hemangioma is difficult but should be kept in mind as being part of the differential diagnosis of adrenal tumors.

Adrenal Gland Neoplasms↗

PubMed1998

Heat shock treatment decreases the mortality of sepsis in rats.

Sepsis is an increasingly common and lethal diagnosis in hospitalized patients. In spite of the advances in antibiotics and medical equipment, the mortality rate has not been improved in the last decade. Recently, heat shock proteins (Hsps) have been well documented to play a self-protective role in almost all living cells under various pathological and physiological stresses through a mechanism known as thermotolerance or cross tolerance. The present study was designed to investigate the expression of Hsp72 and the protective role of pre-induction of Hsps in the mortality during different phases of sepsis. Adult male Sprague-Dawley rats were employed in the study. Sepsis was induced by cecal ligation and puncture (CLP). Heat shock treatment was performed 16 hrs before sepsis induction by heating the rats whole-bodily with an electric heating pad under general anesthesia. The mortality rates with time in both control and preheated groups were monitored. Hsp72 was detected by SDS-PAGE, Western blotting and immunostaining in the brain, heart, liver, kidney, lung, adrenal gland, muscle and lymphocytes. The results show that both early (9 hrs post-CLP) and late (18 hrs post-CLP) sepsis failed to increase the synthesis of Hsp72. Previous induction of Hsps by heat shock treatment significantly decreased the mortality rate of late sepsis. Applying a sufficient inducer to lymphocytes of late sepsis reversed the synthesis of Hsp72 from inactive state into an over-expressive situation in vitro. These results suggest that Hsps are involved in the pathogenesis of sepsis and the involvement of Hsps during the progression of sepsis could add to a first line of host defense against invasive pathogens. Searching for an acceptable agent or less invasive method that leads to increased Hsps expression may provide a means for better treatment of severe infection.

Animals↗

PubMed1997

Retinoid X and retinoic acid receptors interact with transcription factor II-B by distinct mechanisms.

Nuclear hormone receptors are believed to modulate target gene expression by interacting with the general transcriptional machinery of the cell. We demonstrate here that two otherwise closely related members of the nuclear hormone receptor family, retinoid acid receptors (RARs) and retinoid X receptors (RXRs), exhibit significant differences in their interactions with the transcriptional machinery. RARs display a strong constitutive interaction with transcription factor II-B (TFIIB) that requires the TFIIB C-terminus, whereas RXR exhibits a weaker, hormone-stimulated interaction with the TFIIB that maps outside of the TFIIB C-terminus. Use of a dominant-negative mutant of TFIIB suggests that the TFIIB interaction is essential for full transcriptional activation by RXR.

Animals↗

PubMed1997

The RecD subunit of the RecBCD enzyme from Escherichia coli is a single-stranded DNA-dependent ATPase.

We have expressed the RecD subunit of the RecBCD enzyme from Escherichia coli as a fusion protein with a 31-amino acid NH2-terminal extension including 6 consecutive histidine residues (HisRecD). The overexpressed fusion protein can be purified in urea-denatured form by metal chelate affinity chromatography. The mixture of renatured HisRecD protein and the RecB and RecC proteins has a high level of ATP-dependent nuclease activity with either single- or double-stranded DNA, enhanced DNA unwinding activity, enhanced ATP hydrolysis activity in the presence of a small DNA oligomer cosubstrate, and chi-cutting activity. These are all characteristics of the RecBCD holoenzyme. The HisRecD protein by itself hydrolyzes ATP in the presence of high concentrations of single-stranded DNA (polydeoxythymidine). The activity is unstable at 37 degrees C, but is measurable at room temperature (about 23 degrees C). The HisRecD has very little ATPase activity in the presence of a much shorter single-stranded DNA (oligodeoxy(thymidine)12). HisRecD hydrolyzes ATP more efficiently than GTP and UTP, and has very little activity with CTP. We also purified a fusion protein containing a Lys to Gln mutation in the putative ATP-binding site of RecD. This mutant protein has no ATPase activity, indicating that the observed ATP hydrolysis activity is intrinsic to the RecD protein itself.

Adenosine Triphosphatases↗

PubMed1997

Plasticity of tetramer formation by retinoid X receptors. An alternative paradigm for DNA recognition.

Retinoid X receptors (RXRs) are transcription factors that traditionally have been thought to bind DNA as protein dimers. Recently, however, it has been recognized that RXRs can also bind to DNA as protein tetramers. Receptor tetramers form cooperatively on response elements containing suitably reiterated half-sites, and play an important role in determining the specificity of DNA recognition by different nuclear receptors. We report here that RXR tetramers exhibit significant functional plasticity, and form on response elements possessing diverse half-site orientations and spacings. This ability of RXRs to form tetramers and related oligomers appears to contribute to the synergistic transcriptional activation observed when multiple, spatially separated response elements are introduced into a single promoter. Oligomerization may therefore be a common paradigm for DNA recognition and combinatorial regulation by several different classes of transcription factors.

Animals↗

PubMed1997

Vitamin E protection of cell morphology under oxidative stress is related to cytoskeletal proteins in rat hepatocytes.

A significant change in cell morphology was observed in hepatocytes treated with t-butyl hydroperoxide (t-BH). This morphological change of multiple bleb formation on cell plasma membranes was related to cell damage, and the subsequent rupture of these blebs resulted in cell death. In cells incubated with alpha-tocopherol before t-BH treatment, bleb formation was significantly inhibited. Using fluorescence microscopy, actin organization was shown to be related to alpha-tocopherol status as demonstrated by early changes in the actin network of cells in the absence of alpha-tocopherol. Results from SDS-polyacrylamide gel electrophoresis further indicated that, under oxidative stress, actin molecules (45 kDa) decreased in amount and were accompanied by the formation of high molecular weight molecules. In the presence of the thiol reducing agent, dithiothreitol, both the decrease in monomeric actin and formation of high molecular weight molecules disappeared. The loss of actin showed a time-dependent response and could be observed after 15 min with t-BH treatment either in the presence or absence of alpha-tocopherol; however the extent was much more significant in cells with no alpha-tocopherol. Depletion of total membrane protein thiols was also related to vitamin E and was greater in cells with no alpha-tocopherol. The amount of cell damage, as determined by lactate dehydrogenase (LDH) leakage in cells with t-BH treatment over 120 min was decreased in the presence of alpha-tocopherol compared with the rapid increase of LDH leakage in the absence of alpha-tocopherol. These results indicate that vitamin E protection of cell morphology under oxidative stress is related to actin, with thiol groups in actin probably playing a key role.

Actins↗

PubMed1997

No inhibition of gamma-glutamyl transpeptidase-positive foci by vitamin E with or without phenobarbital.

The effect of vitamin E on gamma-glutamyl transpeptidase-positive foci, with or without phenobarbital, was investigated. Groups of six female Sprague-Dawley rats were initiated with diethylnitrosamine (15 mg/kg) at 24 hours of age. After weaning, they were fed diets with 10% (wt/wt) fish oil; the diets contained 0, 5,000 or 15,000 ppm vitamin E supplementation with or without phenobarbital (500 ppm) for six months. Phenobarbital significantly increased liver weight and liver weight as a percentage of body weight (p < 0.05), suggesting a liver hypertrophic effect of phenobarbital. Phenobarbital significantly decreased hepatic phospholipid arachidonate, eicosapentaenoate, and docosahexaenoate (p < 0.05); this may indicate that phenobarbital stimulates phospholipase A2 activity and results in the increased release of polyunsaturated fatty acids from phospholipids and the decrease of hepatic phospholipid polyunsaturated-to-saturated fat ratio. In rats fed phenobarbital, hepatic vitamin E content was lower than in rats fed no phenobarbital; this suggests that phenobarbital causes oxidative stress or induces enzymes that metabolize the vitamin. Phenobarbital exposure significantly increased hepatic prostaglandin F2 alpha and glutathione S-transferase activity (p < 0.05). Vitamin E did not influence hepatic gamma-glutamyl transpeptidase-positive foci area and number with or without phenobarbital, and phenobarbital showed a strong promoting action on enzyme-altered hepatic foci.

Animals↗

PubMed1997

Corn oil enhances gamma-glutamyl transpeptidase-positive foci development in the presence of phenobarbital during hepatocarcinogenesis in rats.

We investigated the effects of type of dietary fat and phenobarbital on gamma-glutamyl transpeptidase-positive foci development. Four groups of six female Sprague-Dawley rats were initiated with diethylnitrosamine (15 mg/kg) at 24 hours of age. After weaning, they were fed nutritionally complete semipurified diets containing 15% corn oil or 5% corn oil + 10% fish oil and supplemented with 5,000 ppm vitamin E with or without phenobarbital (500 ppm) for three months. Dietary fish oil significantly increased hepatic phospholipid eicosapentaenoate and docosahexaenoate concentrations and decreased arachidonate concentration compared with 15% corn oil (p < 0.05). Corn oil (15%) significantly increased hepatic prostaglandin F2 alpha concentration compared with 10% fish oil (p < 0.05). Phenobarbital significantly stimulated glutathione S-transferase activity in both dietary fat groups (p < 0.05). In the absence of phenobarbital, type of dietary fat showed no effect on hepatic gamma-glutamyl transpeptidase-positive foci development. However, in the presence of phenobarbital, 15% corn oil significantly enhanced gamma-glutamyl transpeptidase-positive foci development compared with 10% fish oil (p < 0.05). Phenobarbital showed a strong tumor-promoting action in both dietary groups. In conclusion, there was an interaction between type of dietary fat and phenobarbital on gamma-glutamyl transpeptidase-positive foci development during hepatocarcinogenesis in rats.

Animals↗

PubMed1997

Amount and type of dietary lipid modulate rat hepatic cytochrome P-450 activity.

The influence of the amount and type of dietary lipid on rat hepatic cytochrome P-450 activities in the presence and absence of inducer administration was investigated. Weanling male Sprague-Dawley rats were fed fat-free or 20% beef tallow, olive oil, corn oil, linseed oil, or menhaden oil diets in combination with one of the following three treatments: no inducer, intraperitoneal injection of phenobarbital (75 mg/kg body wt) for three consecutive days before they were killed, or intragastric administration of acetone (5 ml/kg) one day before they were killed. Twenty percent linseed oil and menhaden oil diets induced the highest level of activity among the different fat types in the presence of phenobarbital and acetone. Cytochrome P-450IIB1 activity was induced to a significantly greater extent by acetone administration in conjunction with the 20% menhaden oil diet than in conjunction with the other dietary oils (p < 0.05). In the presence of acetone, 20% beef tallow, 20% linseed oil, and 20% menhaden oil diets significantly induced cytochrome P-450IIE1 activity compared with the fat-free diet (p < 0.05). In conclusion, cytochrome P-450IIB1 and P-450IIE1 activities in rats were significantly increased by specific inducers, and dietary lipid was necessary for this effect. Diets supplemented with linseed and menhaden oils were most effective in inducing this activity.

Animal Nutritional Physiological Phenomena↗

PubMed1997

Methionine and cysteine affect glutathione level, glutathione-related enzyme activities and the expression of glutathione S-transferase isozymes in rat hepatocytes.

Methionine and cysteine are constituents of glutathione. To understand the effects of these two sulfur amino acids on the glutathione (GSH)-dependent detoxification defense system, intracellular GSH and GSH-related enzyme activities, including GSH peroxidase, GSH reductase, GSH S-transferase (GST) and gamma-glutamylcysteine synthetase, were determined. In addition, the expression of three GST isozymes and carbonic anhydrase III (CA III) was examined. Hepatocytes isolated from male Sprague-Dawley rats were cultured with 0.1, 0.3, 0.5 or 1.0 mmol/L each of L-methionine and L-cysteine, for up to 7 d. Cells incubated with 0.5 or 1.0 mmol/L methionine and cysteine had increased intracellular GSH. A twofold increase was observed on d 6 compared with freshly isolated hepatocytes (P < 0.05). However, intracellular GSH was lower in cells treated with 0.3 or 0.1 mmol/L each of methionine and cysteine than in cells tested with 0.5 or 1.0 mmol/L. Although the GSH level differed significantly between cells cultured with 0.3 or 1.0 mmol/L of methionine and cysteine, GSH-related enzymes did not differ at these two concentrations. The activity generally remained constant for the first 24 h, then increased up to d 4. Immunodetection analysis revealed no difference in the level of CA III and GST isoforms, Ya, Yb and Yp, with amino acids each at a concentration of at least 0.3 mmol/L. Yp expression steadily increased up to d 7. Most proteins decreased rapidly after 48 h when cultured with 0.1 mmol/L of methionine and cysteine; however, the Yp level increased up to d 6. In conclusion, results indicate that a twofold increase of intracellular GSH is reached by adding methionine and cysteine at a concentration >0.5 mmol/L to the culture medium. The concentrations of methionine and cysteine for maintaining hepatic GSH are higher than for GSH-related enzyme activity and for GST isoform expression.

Amino Acids, Sulfur↗