scholarly journals Increased oxidation of uroporphyrinogen by an inducible liver microsomal system. Possible relevance to drug-induced uroporphyria

1988 ◽  
Vol 250 (1) ◽  
pp. 161-169 ◽  
Author(s):  
F De Matteis ◽  
C Harvey ◽  
C Reed ◽  
R Hempenius
Keyword(s):  
Chick Embryo ◽  
Drug Metabolism ◽  
Microsomal Fraction ◽  
Marked Decrease ◽  
Rat Hepatocytes ◽  
Drug Induced ◽  
Dependent Mechanism ◽  
Stimulation Of

1. The hypothesis that uroporphyria-inducing drugs stimulate the oxidation of uroporphyrinogen by a microsomal NADPH-dependent mechanism was tested. 2. 3,4,3′,4′-Tetrachlorobiphenyl, a very effective inducer of uroporphyria in chick-embryo hepatocyte cultures, stimulates the NADPH-dependent oxidation of uroporphyrinogen by chick-embryo microsomal fraction in vitro. 3. Two different actions of 3,4,3′,4′-tetrachlorobiphenyl are apparently required for this effect: (a) induction of a microsomal system by treatment in vivo and (b) interaction with the induced microsomal fraction in vitro, producing an oxidizing species. 4. The analogue 2,4,2′,4′-tetrachlorobiphenyl is relatively ineffective in both the production of porphyria in culture and the stimulation of porphyrinogen oxidation in vitro. 5. Rat hepatocytes do not develop uroporphyria when treated with polychlorinated biphenyls in culture, yet they respond to these drugs with typical induction of cytochrome P-448-dependent drug metabolism. 6. These data provide support for the hypothesis of an increased oxidation of uroporphyrinogen in drug-induced uroporphyria, but also suggest that induction of cytochrome P-448 is not the only factor involved. 7. Both I and III isomers of uroporphyrin and heptacarboxylate porphyrin accumulate when chicken hepatocytes are made uroporphyric by drugs; treatment with desferrioxamine causes a marked decrease in both isomers, suggesting that iron may be involved in the accumulation of both.

scholarly journals Stimulation of collagen galactosyltransferase and glucosyltransferase activities by lysophosphatidylcholine

1976 ◽  
Vol 160 (1) ◽  
pp. 29-35 ◽  
Author(s):  
H Anttinen
Keyword(s):  
Chick Embryo ◽  
High Speed ◽  
Membrane Structures ◽  
Embryo Extract ◽  
Chick Embryo Extract ◽  
Triton X ◽  
Regulatory Significance ◽  
Stimulation Of

Lysophosphatidylcholine stimulated the activities of collagen galactosyl- and glucosyl-transferases in chick-embryo extract and its particulate fractions in vitro, whereas essentially no stimulation was noted in the high-speed supernatant, where the enzymes are soluble and membrane-free. The stimulatory effect of lysophosphatidylcholine was masked by 0.1% Triton X-100. In kinetic experiments lysophosphatidylcholine raised the maximum velocities with respect to the substrates and co-substrates, whereas no changes were observed in the apparant Km values. Phospholipase A preincubation of the chick-embryo extract resulted in stimulation of both transferase activities, probably gy generating lysophosphatides from endogenous phospholipids. No stimulation by lysophosphatidylcholine was found when tested with 500-fold-purified glycosyltransferase. The results suggest that collagen glycosyltransferases must be associated with the membrane structures of the cell in order to be stimulated by lysophosphatidylcholine. Lysophosphatidylcholine could have some regulatory significance in vivo, since its concentration in the cell is comparable with that which produced marked stimulation in vitro.

scholarly journals Activation of protein kinase B β and γ isoforms by insulin in vivo and by 3-phosphoinositide-dependent protein kinase-1 in vitro: comparison with protein kinase B α

1998 ◽  
Vol 331 (1) ◽  
pp. 299-308 ◽  
Author(s):  
Kay S. WALKER ◽  
Maria DEAK ◽  
Andrew PATERSON ◽  
Kevin HUDSON ◽  
Philip COHEN ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Protein Kinase ◽  
Protein Kinase B ◽  
Rat Hepatocytes ◽  
Dependent Protein Kinase ◽  
L6 Myotubes ◽  
Dependent Protein ◽  
Stimulation Of

The regulatory and catalytic properties of the three mammalian isoforms of protein kinase B (PKB) have been compared. All three isoforms (PKBα, PKBβ and PKBγ) were phosphorylated at similar rates and activated to similar extents by 3-phosphoinositide-dependent protein kinase-1 (PDK1). Phosphorylation and activation of each enzyme required the presence of PtdIns(3,4,5)P3 or PtdIns(3,4)P2, as well as PDK1. The activation of PKBβ and PKBγ by PDK1 was accompanied by the phosphorylation of the residues equivalent to Thr308 in PKBα, namely Thr309 (PKBβ) and Thr305 (PKBγ). PKBγ which had been activated by PDK1 possessed a substrate specificity identical with that of PKBα and PKBβ towards a range of peptides. The activation of PKBγ and its phosphorylation at Thr305 was triggered by insulin-like growth factor-1 in 293 cells. Stimulation of rat adipocytes or rat hepatocytes with insulin induced the activation of PKBα and PKBβ with similar kinetics. After stimulation of adipocytes, the activity of PKBβ was twice that of PKBα, but in hepatocytes PKBα activity was four-fold higher than PKBβ. Insulin induced the activation of PKBα in rat skeletal muscle in vivo, with little activation of PKBβ. Insulin did not induce PKBγ activity in adipocytes, hepatocytes or skeletal muscle, but PKBγ was the major isoform activated by insulin in rat L6 myotubes (a skeletal-muscle cell line).

scholarly journals Flutamide-Induced Cytotoxicity and Oxidative Stress in anIn VitroRat Hepatocyte System

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Abdullah Al Maruf ◽  
Peter O’Brien
Keyword(s):  
Oxidative Stress ◽  
Tissue Injury ◽  
Rat Hepatocytes ◽  
Experimental Models ◽  
Drug Induced ◽  
Isolated Rat Hepatocytes ◽  
Ros Scavenger ◽  
Generating System

Flutamide (FLU) is a competitive antagonist of the androgen receptor which has been reported to induce severe liver injury in some patients. Several experimental models suggested that an episode of inflammation during drug treatment predisposes animals to tissue injury. The molecular cytotoxic mechanisms of FLU in isolated rat hepatocytes using anin vitrooxidative stress inflammation system were investigated in this study. When a nontoxic hydrogen peroxide (H2O2) generating system (glucose/glucose oxidase) with peroxidase or iron(II) [Fe(II)] (to partly simulatein vivoinflammation) was added to the hepatocytes prior to the addition of FLU, increases in FLU-induced cytotoxicity and lipid peroxidation (LPO) were observed that were decreased by 6-N-propyl-2-thiouracil or deferoxamine, respectively.N-Acetylcysteine decreased FLU-induced cytotoxicity in this system. Potent antioxidants, for example, Trolox ((±)-6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), resveratrol (3,5,4′-trihydroxy-trans-stilbene), and DPPD (N,N′-diphenyl-1,4-phenylenediamine) also significantly decreased FLU-induced cytotoxicity and LPO and increased mitochondrial membrane potential (MMP) and glutathione (GSH) levels in the H2O2generating system with peroxidase. TEMPOL (4-hydroxy-2,2,6,6-tetramethylpiperidin-1-oxyl), a known reactive oxygen species (ROS) scavenger and superoxide dismutase mimetic, also significantly decreased toxicity caused by FLU in this system. These results raise the possibility that the presence or absence of inflammation may be another susceptibility factor for drug-induced hepatotoxicity.

Prostaglandins E2 and F2α are not involved in the monocytic-product (interleukin-1)-induced stimulation of hepatic fibrinogen synthesis in rats

1988 ◽  
Vol 74 (5) ◽  
pp. 477-483 ◽  
Author(s):  
J. C. W. M. Holtslag ◽  
H. J. Moshage ◽  
J. F. van Pelt ◽  
J. A. G. M. Kleuskens ◽  
F. W. J. Gribnau ◽  
...  
Keyword(s):  
Acute Phase ◽  
Acute Phase Response ◽  
Second Messengers ◽  
Interleukin 1 ◽  
Rat Hepatocytes ◽  
Phase Response ◽  
Mrna Content ◽  
Stimulation Of

1. Monocytic products, especially interleukin-1 (IL-1), play an important role in the acute-phase response. Prostaglandins have been shown to act as second messengers in several physiological alterations of the acute-phase response, such as fever, muscle wasting and immunoregulation. The present study was undertaken to determine the role of prostaglandins in the monocytic-product-induced stimulation of the hepatic synthesis of fibrinogen, a well-known acute-phase protein. 2. Prostaglandin (PG) E2, PGF2α and 16,16-dimethyl-PGE2 did not stimulate fibrinogen synthesis and fibrinogen polypeptide mRNA content when administered intraperitoneally to rats or when added to monolayer cultures of rat hepatocytes. 3. Cyclo-oxygenase inhibitors did not abolish the stimulation of fibrinogen synthesis and its mRNA content induced by monocytic products in vivo or in vitro. 4. These findings indicate that the enhanced synthesis of fibrinogen induced by monocytic products (including IL-1) during the acute-phase response is not mediated by prostaglandins or other products of the cyclo-oxygenase pathway of arachidonic acid.

scholarly journals Development of Scaffolds with Adjusted Stiffness for Mimicking Disease-Related Alterations of Liver Rigidity

2020 ◽  
Vol 11 (1) ◽  
pp. 17 ◽  
Author(s):  
Marc Ruoß ◽  
Silas Rebholz ◽  
Marina Weimer ◽  
Carl Grom-Baumgarten ◽  
Kiriaki Athanasopulu ◽  
...  
Keyword(s):  
Drug Metabolism ◽  
Liver Toxicity ◽  
Calf Serum ◽  
Liver Stiffness ◽  
In Vivo Studies ◽  
Drug Induced ◽  
Fibrotic Liver ◽  
Vitro Model

Drug-induced liver toxicity is one of the most common reasons for the failure of drugs in clinical trials and frequent withdrawal from the market. Reasons for such failures include the low predictive power of in vivo studies, that is mainly caused by metabolic differences between humans and animals, and intraspecific variances. In addition to factors such as age and genetic background, changes in drug metabolism can also be caused by disease-related changes in the liver. Such metabolic changes have also been observed in clinical settings, for example, in association with a change in liver stiffness, a major characteristic of an altered fibrotic liver. For mimicking these changes in an in vitro model, this study aimed to develop scaffolds that represent the rigidity of healthy and fibrotic liver tissue. We observed that liver cells plated on scaffolds representing the stiffness of healthy livers showed a higher metabolic activity compared to cells plated on stiffer scaffolds. Additionally, we detected a positive effect of a scaffold pre-coated with fetal calf serum (FCS)-containing media. This pre-incubation resulted in increased cell adherence during cell seeding onto the scaffolds. In summary, we developed a scaffold-based 3D model that mimics liver stiffness-dependent changes in drug metabolism that may more easily predict drug interaction in diseased livers.

ACTION OF THYROTROPHIN (TSH) ON THYROID PROTEIN SYNTHESIS IN VIVO AND IN VITRO

1973 ◽  
Vol 72 (3) ◽  
pp. 453-463 ◽  
Author(s):  
Gustav Wägar ◽  
Ragnar Ekholm ◽  
Ulla Björkman
Keyword(s):  
Amino Acids ◽  
Protein Synthesis ◽  
Free Amino ◽  
Microsomal Fraction ◽  
Inhibitory Effect ◽  
Leucine Incorporation ◽  
Bovine Thyroid ◽  
Stimulation Of

ABSTRACT The effect of TSH on the incorporation of L-14C-leucine into thyroid proteins was studied in vivo in rats as well as in vitro on bovine thyroid slices and a microsomal subfraction. It was found that TSH reduced the incorporation of radio-leucine into the proteins of slices during the first 2 hours when the concentration of non-labelled leucine in the incubation medium was low. When cold leucine was added to the medium this inhibitory effect was no longer observed. After 6 hours a stimulatory effect on the radio-leucine incorporation by TSH was obvious at both low and high leucine concentrations. The incorporation of 14C-leucine into proteins by the microsomal fraction incubated with a pH 5-fraction was reduced by TSH but this inhibitory effect of TSH disapperaed when post-microsomal supernatant, containing free amino acids, was added to the incubation mixture. It is suggested that the apparent inhibitory effect of TSH on protein synthesis in thyroid slices is due to an altered ratio labelled/non-labelled leucine, caused by stimulation of proteolysis by TSH. This explanation does not seem applicable, however, to the similar apparently inhibitory effect of TSH on protein synthesis observed in the microsomal fraction. In the in vivo experiments a stimulation of the incorporation of labelled leucine could not be observed until 4 hours after the TSH administration. It is suggested that this apparently slow effect of TSH on protein synthesis might be explained either by an indirect effect of TSH on protein synthesis or by a TSH-induced change of the ratio labelled/non-labelled leucine.

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