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Biomedical subjects

P Linko

Publications and source records attributed to P Linko.

At least 37 records · Page 2Linked to original sources
PubMed1991

Glutathione S-transferase mu in human lymphocyte and liver: role in modulating formation of carcinogen-derived DNA adducts.

Glutathione transferase (GT) activity towards trans-stilbene oxide (tSBO), benzo[a]pyrene-4,5-oxide (B[a]PO) and 1-chloro-2,4-dinitrobenzene (CDNB) was measured in human liver and lymphocytes. GT-tSBO activity is catalyzed by GT mu which has polymorphic expression in human lymphocytes. Our results show that activity of GT-tSBO in lymphocytes correlates with its activity in liver (r = 0.7, P less than 0.001). GT activity towards BPO (GT-BPO) also correlated with GT-tSBO in lymphocytes and liver. However, interindividual variation of GT-BPO is less than that of GT-tSBO, suggesting that BPO may not be as specific a substrate for GT mu and therefore other GT isozymes may contribute to BPO conjugation. Conjugation of CDNB by GT was not different using cytosols from either high or low GT mu individuals. The functional significance of the GT-mu polymorphism was evaluated by measuring its effect on benzo[a]pyrene (B[a]P)- and aflatoxin B1 (AFB1)-DNA adduct formation in vitro. Human liver cytosols prepared from persons having low or high GT-tSBO activity were incubated with human liver microsomes, calf thymus DNA and B[a]P or AFB1. HPLC analysis revealed that the major B[a]P adduct was dG(N2)-7 beta, 8 alpha-trihydroxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BPDE-dG). BPDE-dG adducts were decreased equally by cytosols from either low or high conjugators. In contrast, AFB1-DNA binding was inhibited to a greater extent in high conjugators than low conjugators. HPLC analysis demonstrates that adducts formed were AFB1-FAPyr and AFB1-N7-Gua. The correlation between AFB1-DNA adduct concentrations and GT mu activity was highly significant with a correlation coefficient of r = 0.88 at P less than 0.001. These results suggest that GT mu plays an important role in detoxifying DNA reactive metabolites of AFB1 and this enzyme may be a susceptibility marker for AFB1 related liver cancer. Moreover, our data demonstrate that lymphocytes are a reliable surrogate tissue for detecting liver GT mu polymorphisms.

Adolescent↗
PubMed1989

Hormonal regulation of male-specific rat hepatic cytochrome P-450g (P-450IIC13) by androgens and the pituitary.

The present study examines the hormonal regulation male-specific cytochrome P-450g (IIC13) and its mRNA. Neonatal gonadectomy of male rats abolished hepatic expression of P-450g and its mRNA in adulthood, while ovariectomy had little effect. Neonatal administration of testosterone to neonatally gonadectomized male or female rats partially masculinized expression of P-450g and its mRNA, and postpubertal administration of testosterone (testosterone capsules implanted at 5 weeks) completely masculinized their expression. However, castration of male rats at puberty (5 weeks) had no effect on P-450g or its mRNA at 10 weeks. Male-specific development of P-450g and P-450 M-1 (IIC11) mRNA were imprinted similarly by testosterone. However, hypophysectomy experiments demonstrated that the two male-specific forms of P-450 are regulated quite differently. Hypophysectomy of male rats decreased hepatic content of P-450 M-1 mRNA by approximately 50%, and intermittent injections of growth hormone completely restored this mRNA. In contrast, hypophysectomy of male rats increased P-450g and its mRNA by approximately 50%, while intermittent injections of growth hormone produced a slight decrease. Hypophysectomy of female rats increased P-450g and its mRNA to adult male levels, but produced only a small increase in P-450 M-1 mRNA. Continuous infusion of growth hormone into sham hypophysectomized male rats (to mimic the female growth hormone pattern) resulted in a complete loss of P-450g and its mRNA. These results indicate that the expression of P-450g is not dependent on the male pulsatile growth hormone pattern, but suggest instead that the continuous secretion of growth hormone suppresses P-450g in the female rat.

Animals↗
PubMed1989

Interaction of hexachlorobenzene with the receptor for 2,3,7,8-tetrachlorodibenzo-p-dioxin in vitro and in vivo. Evidence that hexachlorobenzene is a weak Ah receptor agonist.

Hexachlorobenzene (HCB) produces hepatic porphyria and induces the hepatic cytochrome P450 isozymes P450c (P450IA1) and P450d (P450IA2) in rodents. These and other effects of HCB resemble those of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), which acts via its binding to the aromatic hydrocarbon (Ah) receptor. We therefore examined the ability of HCB to interact with this receptor in vitro and in vivo. HCB, at concentrations of 1 microM or higher, inhibited the specific binding of [3H]TCDD (0.3 nM) to the Ah receptor in vitro, whereas the solubility of [3H]TCDD was affected only at 100 microM HCB. The inhibition was competitive, with a KI of approximately 2.1 microM. In rats fed a diet containing 3000 ppm HCB for varying times (4 h to 7 days), the specific binding of [3H]TCDD in hepatic cytosol was reduced by up to 40%, as observed previously for known Ah receptor agonists. The decrease in [3H]TCDD specific binding in cytosol of HCB-treated rats was due principally to a decrease in the number of binding sites for [3H]TCDD rather than competition from residual HCB. As shown by immunoblotting and radioimmunoassay, HCB induced the cytochrome P450 isozymes P450c and P450d, which are regulated by the Ah receptor, as well as the phenobarbital-inducible isozymes P450b and P450e. Together these results indicate that HCB is a weak agonist for the Ah receptor, and suggest that some of its effects may be mediated by its interaction with this gene-regulatory protein.

Animals↗
PubMed1989

Suppression of male-specific cytochrome P450 2c and its mRNA by 3,4,5,3',4',5'-hexachlorobiphenyl in rat liver is not causally related to changes in serum testosterone.

Rat cytochrome P450 2c (P450 gene IIC11) is a constitutive, male-specific hepatic enzyme which is suppressed greater than 90% by treatment with 3,4,5,3',4',5'-hexachlorobiphenyl (HCB) [H. N. Yeowell et al. (1987) Mol. Pharmacol. 32, 340-347]. HCB also decreases serum testosterone levels in adult male rats (greater than 98% loss). The present study assesses whether the suppression of P450 2c by HCB is a direct result of its effects on serum testosterone levels. Further, the site along the hypothalamic-pituitary-testicular axis at which HCB acts to depress testosterone secretion was examined. Administration of the synthetic androgen methyltrienolone to HCB-treated rats failed to prevent the suppression of P450 2c mRNA and its associated microsomal steroid 16 alpha-hydroxylase activity under conditions where it effectively reversed the large decrease in P450 2c mRNA and steroid 16 alpha-hydroxylase activity produced by castration. Hepatic steroid 6 beta-hydroxylase activity, which is catalyzed primarily by P450 2a (P450 gene IIIA2), was also suppressed by HCB and was not protected by methyltrienolone. Administration of either human chorionic gonadotropin, an analog of pituitary-derived luteinizing hormone, or the hypothalamic luteinizing hormone releasing hormone elevated serum testosterone levels to a much smaller extent in HCB-treated rats than in control rats. These results indicate that the effects of HCB on serum testosterone levels reflect its effects on testicular function rather than the pituitary or hypothalamus. However, the present study demonstrates that the consequential reduction in serum testosterone levels in HCB-treated rats is not causally related to the reduction in hepatic P450 2c levels. Thus, HCB must also act on some other regulatory mechanism involved in the expression of this protein.

Animals↗
PubMed1988

The role of the Ah locus in hexachlorobenzene-induced porphyria. Studies in congenic C57BL/6J mice.

The role of the Ah locus in hexachlorobenzene (HCB)-induced porphyria and the possible involvement of P-450 cytochromes P(1)450 and P(3)450 in the pathogenesis of this disease were investigated in two congenic strains of C57BL/6J mice that differ only at this locus. Female B6-Ahb mice (Ah receptor: approximately 30-70 fmol/mg of cytosolic protein) and B6-Ahd mice (Ah receptor: undetectable) were pretreated with iron (500 mg/kg) and then fed a diet containing 0 or 200 p.p.m. of HCB for up to 17 weeks. Mice from the two strains consumed similar amounts of HCB. Urinary excretion of porphyrins was increased after 7 weeks of HCB treatment in B6-Ahb mice, and after 15 weeks was over 200 times greater than that of mice given iron only. In B6-Ahd mice, porphyrin excretion did not begin to increase until after 13 weeks, and after 15 weeks was only six times greater than that of controls. Similar differences were seen in the 15-week hepatic porphyrin concentrations (B6-Ahb: 1110 +/- 393; B6-Ahd: 17.6 +/- 14.5; controls: approximately 0.20 nmol/g). Uroporphyrinogen decarboxylase (EC 4.1.1.37) activity was diminished by 70 and 20% in B6-Ahb B6-Ahd mice respectively after 15 weeks of treatment with HCB. Cytochromes P(1)450 and P(3)450 were measured in hepatic microsomes (microsomal fractions) by radioimmunoassay and immunoblotting, using antisera raised against the orthologous rat isoenzymes P450c and P450d. HCB induced small amounts of a protein recognized by anti-P450c (P(1)450) in B6-Ahd mice, but not in B6-Ahd mice. Relatively large amounts of a protein recognized by anti-P450d (P(3)450) were induced in both strains, but to a somewhat greater extent in the B6-Ahb mice. The hepatic accumulation of HCB at 15 weeks was greater in B6-Ahb than in B6-Ahd mice, in association with elevated hepatic lipid levels in the former strain. The results of this experiment indicate that the Ah locus influences the susceptibility of C57BL/6J mice to HCB-induced porphyria and are consistent with the suggestion that the sustained induction of P(3)450 and/or P(1)450 may be a causative factor in the development of this disease.

Animals↗
PubMed1988

Increases in cytochrome P-450 mediated 17 beta-estradiol 2-hydroxylase activity in rat liver microsomes after both acute administration and subchronic administration of 2,3,7,8-tetrachlorodibenzo-p-dioxin in a two-stage hepatocarcinogenesis model.

Administration of single doses of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) (10 micrograms/kg) increased estradiol 2-hydroxylase (E2OHase) activity approximately 2-fold in liver microsomes of female rats but had no effect on E2OHase activity in hepatic microsomes of male rats. In contrast, TCDD increased P-450d (an enzyme which has a high turnover number for E2OHase in a reconstituted enzyme system) 10- to 20-fold in livers of both male and female rats. The discrepancy between the increases in P-450d and E2OHase activity in liver microsomes of TCDD-induced rats was abolished by the addition of exogenous purified P-450 reductase to the microsomal assays for E2OHase, suggesting that reductase was limiting in the in vitro assays. When E2OHase activity was assayed in the presence of exogenous reductase, TCDD increased E2OHase 2-fold and 4-fold respectively in liver microsomes of male and female rates. Antibody to P-450d completely inhibited the increase in E2OHase activity in liver microsomes of TCDD-treated male and female rats, but had little effect on E2OHase activity in liver microsomes of untreated male or female rats. These data indicate that P-450d is responsible for the increase in E2OHase activity in TCDD-treated rats, but other P-450 isozymes are responsible for constitutive E2OHase activity. Biweekly administration of 1.4 micrograms/kg of TCDD for 30 weeks as a potential promoter of hepatocarcinogenesis increased the volume of the liver occupied by gamma-glutamyl transpeptidase (GGT)-positive foci in livers of female rats given a necrogenic dose of diethylnitrosamine (DEN) (200 mg/kg) as the initiator. Biweekly doses of 0.14-1.4 micrograms/kg TCDD in this model also increased P-450d (7-fold) and E2OHase activity maximally (4-fold) in DEN-initiated rats. Moreover, initiation with DEN substantially enhanced the effects of the low dose of TCDD on both hepatic microsomal P-450d and E2OHase activity.

Animals↗
PubMed1988

Induction of rat cytochrome P-450 3 and its mRNA by 3,4,5,3',4',5'-hexachlorobiphenyl.

Rat cytochrome P-450 3 (P-450 3) is a constitutive hepatic steroid hormone 7 alpha-hydroxylase which is relatively unresponsive to a number of monooxygenase-inducing agents. The present study demonstrates that a polyhalogenated aromatic hydrocarbon inducer, 3,4,5,3',4',5'-hexachlorobiphenyl (HCB), induces P-450 3 in livers of adult male rats, and that the increase is the result of an increase in the mRNA for this enzyme. Cytochrome P-450 3 and its mRNA were increased more slowly and to a lesser extent than cytochrome P-450c (P-450c) and its mRNA, indicating that these enzymes are not regulated coordinately in liver. The maximum increase in P-450 3 and P-450 3-dependent androstenedione 7 alpha-hydroxylase activity (2- to 3-fold) occurred 7 days after administration of HCB, in contrast to the increase in P-450c (greater than 200-fold) which was maximal by 3-5 days. The rate of induction of P-450 3 mRNA was also slower [maximum increase (9-fold) at 5 days after HCB administration] than that of P-450c mRNA [maximum increase (30-fold) at 2-3 days]. Moreover, a higher dose of HCB was required to produce maximum induction of P-450 3 (50 mg/kg) than that required to produce maximum induction of P-450c (10 mg/kg). P-450 3 was not detected on Western blots of lung, kidney, or prostate microsomes isolated from control or HCB-treated rats (less than or equal to 2% of that found in livers of HCB-treated rats). Moreover, P-450 3-dependent steroid 7 alpha-hydroxylase activity was not detected in these extrahepatic tissues of control or HCB-treated rats (less than or equal to 1% of that found in the corresponding liver microsomes of untreated or HCB-treated rats). In contrast, P-450c was increased dramatically by HCB in lung, kidney, and prostate tissues, indicating differential expression of P-450c and P-450 3 in extrahepatic tissues.

Animals↗
PubMed1986

Induction of cytochrome P-450 isozymes by hexachlorobenzene in rats and aromatic hydrocarbon (Ah)-responsive mice.

Hexachlorobenzene (HCB) differs markedly from other chlorinated benzenes (CBs) as an inducer of cytochrome P-450 (P-450) isozymes as determined by radioimmunoassay and immunoblotting. At greater than 99% pure, HCB induced both the phenobarbital-inducible forms, cytochromes P-450b + e (70 chi), and the 3-methylcholanthrene-inducible forms, cytochromes P-450c (58 chi) and P-450d (8 chi), in rat liver microsomes. The concentration of P-450d was considerably greater than that of P-450c in HCB-induced rat liver. In contrast to HCB, all lower chlorinated benzenes tested were PB-type inducers. Hexachlorobenzene increased the amounts of translatable messenger RNAs (mRNAs) for P-450b, P-450c, and P-450d in rat liver polysomes, suggesting that it increases the synthesis of these proteins. Evidence that HCB interacted with the putative Ah receptor for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) was equivocal. Western blots of liver microsomes from Ah-responsive C57BL/6J (B6) and nonresponsive DBA/2J (D2) mice demonstrated that HCB produced a large increase in P3-450 and a very small increase in P1-450 in the responsive strain. The increase in P1-450 was not observed after HCB administration to nonresponsive mice, but a small increase in P3-450 was noted. These findings suggested that HCB may act through the Ah receptor. However, HCB was at best a very weak competitor for specific binding of [3H]-TCDD to the putative receptor in rat or mouse hepatic cytosol in vitro, producing decreases in binding of [3H]-TCDD only at very high concentrations (10(-6) to 10(-5) M).

Animals↗
PubMed1986

Structure-activity relationships of chlorinated benzenes as inducers of hepatic cytochrome P-450 isozymes in the rat.

This study compared the ability of hexachlorobenzene (HCB) and of other chlorinated benzenes to induce cytochrome P-450 isozymes in rat liver. HCB (greater than 99% pure) induced both the phenobarbital-inducible forms (cytochrome P-450b and P-450e) and the 3-methylcholanthrene (3-MC)-inducible forms (P-450c and P-450d) of cytochrome P-450. However, HCB differed from many 3-MC-type inducers by inducing P-450d preferentially over P-450c. In contrast to HCB, the lower chlorinated benzenes did not induce significant amounts of P-450c or P-450d in the rat, but were phenobarbital-type inducers, inducing P-450b and P-450e. These data indicate that the hepatic effects of HCB differ markedly from those of other chlorinated benzenes. However, chlorinated dibenzodioxins also induce P-450c and P-450d in the rat, and although chlorinated dibenzodioxins and dibenzofurans contaminate certain commercial products, none were detected by gas chromatography/mass spectrometry (detection limit 0.5 ppm) in the HCB used in this study. The evidence that HCB interacted with the receptor for 2,3,7,8-tetrachlorodibenzo-para-dioxin (TCDD) was equivocal. At a concentration of 10(-6) M, HCB produced a slight decrease (18%) in the binding of 3H-TCDD to this protein in vitro, but had no effect at lower concentrations. However, as an inducer of two 3-MC-inducible isozymes of P-450, HCB was clearly more effective in aromatic-hydrocarbon-responsive mice (C57Bl/6J) than in non-responsive mice (DBA/2J), suggesting that HCB may act through the Ah receptor.

Animals↗
PubMed1985

Induction of specific cytochrome P-450 isozymes by methylenedioxyphenyl compounds and antagonism by 3-methylcholanthrene.

Two methylenedioxyphenyl compounds, isosafrole (5-propenyl-1,3-benzodioxole) and an analog, 5-t-butyl-1,3-benzodioxole (BD), differ markedly as inducers of cytochrome P-450 isozymes in rat liver microsomes. Isosafrole is a mixed-type inducer, inducing P-450b, P-450c, and P-450d. In contrast, BD is a phenobarbital-type inducer, increasing P-450b, but producing little or no increase in P-450c or P-450d. Similarly, isosafrole increases the amount of translatable mRNA for P-450b, c and d, while BD induces only the mRNA for P-450b. Dimethylation of the methylene bridge carbon of BD to give 2,2-dimethyl-5-t-butyl-1,3-benzodioxole (DBD) blocks the formation of NADPH-reduced Type III metabolite-P-450 complexes in vitro, and diminishes but does not abolish the ability of the compound to induce P-450b. Western blots of microsomes from isosafrole and BD-treated rat livers confirm that in contrast to isosafrole, BD does not induce P-450d or P-450c. However, the antibody to P-450d recognizes two new polypeptides (approximately 50K Mr) from sodium dodecyl sulfate-polyacrylamide gels of liver microsomes from BD-treated rats. These polypeptides are not observed in control, isosafrole, 3-methylcholanthrene (3-MC), or DBD-treated rats. They are intensified by coadministration of 3-MC with BD and may represent either modified isozyme-metabolite adducts or degradation products of P-450d. However, the polypeptides could not be generated in vitro by addition of BD to 3-MC-induced microsomes with NADPH under conditions which produced spectral metabolite complexes, or in a reconstituted system with P-450d. The two methylenedioxyphenyl compounds do not form stable metabolite complexes with the same P-450 isozymes. BD formed distinct spectral metabolite complexes in vitro with both P-450b and P-450c but not with P-450d in a reconstituted system. In contrast, isosafrole forms metabolite complexes with all three isozymes. Coadministration of 3-MC with BD blocked induction of P-450b by 80% and produced a similar repression of its translatable mRNA. This finding indicates that 3-MC type inducers not only induce certain cytochrome P-450 isozymes, but also repress synthesis of other isozymes.

Animals↗
PubMed1985

Fuels and industrial chemicals through biotechnology. 2.

An overview of research on the biotechnical production of fuels and industrial chemicals during the two-year period of 1983-1984 is presented. Ethanol fermentation has continued to be the subject of major interest. A considerable amount of work has been directed to alternative feedstocks such as pentose sugars and lactose, and to bacterial fermentations. Reports on extrusion cooking as a continuous pretreatment method for subsequent ethanol fermentation, and on novel alternative downstream processing techniques have been published. In addition to ethanol fermentation, much attention has been paid to the biotechnical production of 2,3-butanediol, and of a number of organic and amino acids. In general, there appears to be a growing interest in the application of biocatalysis for the production of specialty chemicals, although only a few examples will be discussed in this paper. The construction of a demonstration plant to produce ethanol from molasses by a two 10 kL bed-volume immobilized yeast bioreactors at the Kyowa Hakko Kogyo Company Hofu plant, the announcement by Nitto Chemical Industries Company to begin the biotechnical production of acrylamide, and the French decision to construct pilot plants for the biotechnical production of acetone-butanol-ethanol cosolvent and of ethanol from renewable resources represent major scale-up developments.

Journal Article↗
PubMed1985

Dose response for induction of two cytochrome P-450 isozymes and their mRNAs by 3,4,5,3'4'5'-hexachlorobiphenyl indicating coordinate regulation in rat liver.

The present study compares the time course and dose-response curves for induction of the two major 3-methylcholanthrene (3-MC)-inducible isozymes of cytochrome P-450 and their mRNAs in livers of male rats after administration of 3,4,5,3'4'5'-hexachlorobiphenyl (HCB). Isozyme concentrations were measured by radioimmunoassay. The corresponding translatable mRNAs were measured by translation of polysomes in a cell-free translational system followed by immunoprecipitation and electrophoretic analysis of the translational products. The time course for induction of the two isozymes by HCB indicated that cytochrome P-448MC (P-450c) peaked sooner than P-448HCB (P450d). However, the time course for induction of the two mRNAs was identical. The dose-response curves for induction of the two isozymes and their mRNAs demonstrated that the ED50 for induction of P-448MC was identical to that of P-448HCB, suggesting that the two proteins are induced coordinately by this compound in liver. HCB did not induce P-450PB (the major phenobarbital-inducible isozyme) or affect mRNA levels for this isozyme. Although cytochrome P-448HCB is the predominant cytochrome in liver microsomes from HCB-induced rats, the magnitude of the induction of this isozyme (40-fold) is lower than that of P-448MC (600-fold), because cytochrome P-448HCB is present in higher concentrations in livers of untreated rats than P-448MC (90 versus 3 pmol/mg). Polysomes from control rats also contain more translationally active P-448HCB mRNA than P-448MC mRNA (0.009 versus 0.003% of the total translational products). The increase in the translatable mRNAs (12-fold for P-448HCB mRNA and 40-fold for P-448MC mRNA) was less than the increase in the isozymes. The discrepancy between the magnitude of the induction of the isozymes and their respective mRNAs suggests that factors other than an increase in mRNA influence the magnitude of the increase of the isozymes by HCB. However, HCB did not affect translational efficiency of total mRNA as measured in vitro in the present study. Differences in half-lives of the proteins or effects of HCB on the stability of the proteins might account for the magnitude of the increase in the isozymes after HCB treatment.

Animals↗
PubMed1984

Differential induction of two 2,3,7,8-tetrachlorodibenzo-p-dioxin-inducible forms of cytochrome P-450 in extrahepatic versus hepatic tissues.

The present study examines the induction of two isozymes of cytochrome P-450, P-448HCB and P-448MC, by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in hepatic and a number of extrahepatic tissues of male rats. These isozymes were quantitated by radioimmunoassay (RIA). TCDD induces both forms of cytochrome P-448 markedly in liver. In extrahepatic tissues, TCDD induces cytochrome P-448MC but not cytochrome P-448HCB. Induction of cytochrome P-448MC is greatest in liver greater than kidney greater than lung greater than intestine greater than spleen greater than testes greater than brain (no significantly increased). The results in liver, kidney, and lung were confirmed by a technique that depends on both electrophoretic mobility and immunological characteristics (sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by transfer to nitrocellulose paper and immunostaining of the protein). Cytochrome P-448HCB is a minor constitutive form in livers of control male rats (5% of the total cytochrome P-450). In contrast, cytochrome P-448MC is below the level of detection in control livers using the immunostaining technique (less than 0.6% by RIA). These results indicate that induction of cytochrome P-450 isozymes by TCDD is tissue-dependent in the rat. The response of extrahepatic tissues to TCDD is more limited than that of liver.

Animals↗
PubMed1983

Fuels and industrial chemicals.

The favorite subject of recent literature on biotechnical processes has been ethanol fermentation. This review covers a number of new technics developed, including immobilized biocatalyst technology and bacterial fermentations. Reference is also made to recent work on whey, starch, inulin, and cellulosic materials as substrates for ethanol production. Renewed interest in acetonebutanol fermentation for solvent and liquid fuel production has also been clearly evident during the last two years. Biotechnical production of organic acids has been considered as an alternative route to chemical feedstocks. New developments in amino acid, methane, hydrogen, and hydrocarbon production, and on hydrocarbon oxidation are also briefly covered.

Journal Article↗
PubMed1983

Immunochemical evidence for two 3-methylcholanthrene-inducible forms of cytochrome P-448 in rat liver microsomes using a double-antibody radioimmunoassay procedure.

Double-antibody radioimmunoassay (RIA) procedures were developed for the determination of two forms of cytochrome P-448 in crude liver microsomes at the picogram level. The antisera used in the RIAs were produced against two forms of cytochrome P-448 isolated from livers of rats treated with 3,4,5,3',4',5'-hexachlorobiphenyl (3,4,5-HCB) and 3-methylcholanthrene (3-MC), referred to as cytochromes P-448HCB and P-448MC, respectively. The results obtained with RIA procedures were consistent with results of radial immunodiffusion analysis, sodium dodecyl sulfate/polyacrylamide gel electrophoresis, and estimation of aryl hydrocarbon hydroxylase (AHH) activity. Utilizing the RIAs, both forms of cytochrome P-448 were found in significant quantities in liver microsomes of rats treated with 3-MC, 3,4,5-HCB, isosafrole, and Aroclor 1254 but only in minute concentrations in untreated and phenobarbital-treated rats.

Animals↗
PubMed1982

Induction of porphyria in the rat by chronic versus acute exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Chronic oral administration of 1 microgram . kg-1 . week-1 of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) to female rats for 16 weeks resulted in hepatic porphyria. In contrast, administration of single oral doses as high as 30 micrograms/kg did not produce porphyria, either acutely or 16 weeks later. Activities of hepatic drug-metabolizing enzymes [aryl hydrocarbon hydroxylase (AHH) and glucuronyl transferase] were increased by chronic oral doses of TCDD as low as 0.01 microgram . kg-1 . week-1. When animals were dosed with TCDD chronically and then allowed to recover for 6 months, AHH and glucuronyl transferase activities returned toward normal (98 and 86% recovery). However, animals showed only partial recovery from TCDD-induced porphyria. Hepatic porphyrin levels did decrease during this period, but urinary porphyrins and the rate-limiting enzyme in porphyrin synthesis, delta-aminolevulinic acid synthetase, remained maximally elevated during the 6-month recovery period. It is concluded that single doses of TCDD do not produce porphyria in the rat, but that TCDD is porphyrogenic when given chronically. Moreover, when TCDD administration is stopped, recovery from the porphyrogenic effects of TCDD is very slow and does not correlate with the biological half-life of TCDD in the rat.

5-Aminolevulinate Synthetase↗