John A. Timbrell

The in vivo and in vitro protective properties of taurine

1. Taurine is a ubiquitous, free amino acid found in mammalian systems. 2. The biological functions of taurine are unclear. 3. Various in vivo data suggest that taurine has a variety of protective functions and deficiency leads to pathological changes. 4. Depletion in rats of taurine increases susceptibility to liver damage from carbon tetrachloride. 5. Susceptibility to a variety of hepatotoxicants correlates with the estimated hepatic taurine level. 6. In vitro data suggest that taurine can protect cells against toxic damage. 7. Taurine protects isolated hepatocytes against carbon tetrachloride, hydrazine and 1,4-naphthoquinone but not against allyl alcohol, alpha-naphthylisothiocyanate (ANIT) or diaminodiphenyl methane (DAPM) cytotoxicity. 8. The mechanisms of protection are unclear but may include modulation of calcium levels, osmoregulation and membrane stabilization.

Biomarkers in toxicology

The use of biomarkers in toxicology is becoming increasingly important. This article briefly reviews some of the aspects in an attempt to give an overall view of the field. Some of the new developments, particularly in relation to biomarkers of exposure and response, are mentioned. Specific DNA and protein adducts can now be used as biomarkers of the effective exposure so incorporating variations in environmental levels and individual disposition. Analysis of urinary metabolite profiles by NMR can highlight novel markers and allow recognition of patterns of metabolite changes as biomarkers of a toxic response. Novel urinary markers for liver and testicular dysfunction are discussed. Finally, the acetylator phenotype as a biomarker of susceptibility is described.

Introduction to toxicology

Disposition of Toxic Compunds Types of Exposure and Response Drugs as Toxic Substances Industrial Chemicals Food Additives and Contaminants Pesticides Environmental Pollutants Natural Products Household Products Toxicity Testing and Risk Assessment.

Biochemical characterisation of para-aminophenol-induced nephrotoxic lesions in the F344 rat.

The acute biochemical effects of the nephrotoxin p-aminophenol (PAP) were studied in detail using a combination of conventional bioanalytical and 1H-NMR spectroscopic methods. Dosing PAP (25–100 mg/kg) to male F344 rats resulted in a dose-related proximal nephropathy with consequent elevations in urinary enzymes, glucose, and urine total protein as shown by conventional methodology. 1H-NMR spectroscopy at 400 MHz of urine from PAP-treated rats also revealed a characteristic glycosuria, with concomitant amino aciduria. The increased excretion of these compounds indicates functional defects in the proximal tubule and reduced solute reabsorption efficiency. In addition, 1H-NMR urinalysis and conventional enzymatic analysis showed a dose-related lactic aciduria. Other changes detected by 1H-NMR included a dose-related reduction in the excretion of citrate (confirmed by a conventional biochemical method) and an increase in the excretion of acetate. The degree of abnormalities shown by 1H-NMR urinalysis agreed well with histopathological observations and conventional biochemical indices of nephrotoxicity. 1H-NMR urinalysis therefore serves to highlight changes in the excretion of low MW urine components not routinely studied by conventional biochemical analysis.

Taurine: protective properties against ethanol-induced hepatic steatosis and lipid peroxidation during chronic ethanol consumption in rats.

SummaryAlcohol was administered chronically to female Sprague Dawley rats in a nutritionally adequate totally liquid diet for 28 days. This resulted in hepatic steatosis and lipid peroxidation. Taurine, when co-administered with alcohol, reduced the hepatic steatosis and completely prevented lipid peroxidation. The protective properties of taurine in preventing fatty liver were also demonstrated histologically. Although alcohol was found not to affect the urinary excretion of taurine (a non-invasive marker of liver damage), levels of serum and liver taurine were markedly raised in animals receiving alcohol + taurine compared to animals given taurine alone. The ethanol-inducible form of cytochrome P-450 (CYP2E1) was significantly induced by alcohol; the activity was significantly lower than controls and barely detectable in animals fed the liquid alcohol diet containing taurine. In addition, alcohol significantly increased homocysteine excretion into urine throughout the 28 day period of ethanol administration; however, taurine did not prevent this increase. There was evidence of slight cholestasis in animals treated with alcohol and alcohol + taurine, as indicated by raised serum bile acids and alkaline phosphatase (ALP). The protective effects of taurine were attributed to the potential of bile acids, especially taurine conjugated bile acids (taurocholic acid) to inhibit the activity of some microsomal enzymes (CYP2E1). Thesein vivo findings demonstrate for the first time that hepatic steatosis and lipid peroxidation, occurring as a result of chronic alcohol consumption, can be ameliorated by administration of taurine to rats.

Studies on the relationship between acute testicular damage and urinary and plasma creatine concentration

A single dose of cadmium chloride (3.23 μmol Cd2+/kg) causing acute testicular damage in male rats also caused significant creatinuria and creatinaemia at 48 h after dosing. Doses of cadmium which did not cause testicular necrosis did not cause creatinuria or creatinaemia. Surgical ligation of the pampiniform plexus also caused ischaemic necrosis of the testis and this was followed by significant creatinuria and creatinaemia. However, neither orchidectomy followed by a toxic dose of cadmium, orchidectomy alone nor sham operation caused significant creatinuria or creatinaemia. Cadmium dosing induced a temporary loss of body weight which was less than that caused by food restriction. Food restriction did not cause significant creatinuria but did cause significant creatinaemia. These data suggest that the creatine is derived from the damaged testis and that measurement of urinary creatine may be a useful non-invasive means of detecting acute testicular damage caused by exposure to chemicals or mechanical impairment of blood flow.

Effect of various non-hepatotoxic compounds on urinary and liver taurine levels in rats

Administration of compounds which alter protein synthesis or sulphur amino acid metabolism in rats results in changes in the excretion of urinary taurine. Treatment with diethylmaleate (DEM) or phorone, which will deplete glutathione (GSH), reduces taurine excretion, whereas treatment with buthionine sulphoximine (BSO), which will inhibit glutathione synthesis, increases taurine excretion. Treatment with cycloheximide, an inhibitor of protein synthesis, increases taurine excretion, whereas pretreatment with phenobarbital, which will increase protein synthesis, decreases taurine excretion. Administration of agents which damage organs other than the liver such as the kidney, heart and testes, does not increase urinary taurine.

The effect of age on paracetamol hepatotoxicity in mice

Abstract The hepatotoxicity of paracetamol to mice was investigated in relation to age. Measurement of hepatic necrosis, covalent binding of radiolabelled paracetamol to liver protein and hepatic glutathione depletion indicated that paracetamol was less toxic to neonatal mice than to the adult animal. Although the neonatal hepatic glutathione level is considerably less than that of the adult (26 per cent) the levels of P-450 in the neonate are also less (9 per cent). This suggests that in this case the development of the ability to detoxify the toxic reactive metabolite of paracetamol precedes the development of the enzyme system producing it.

Methionine synthase activity and sulphur amino acid levels in the rat liver tumour cells HTC and Phi-1

Methionine dependence has been reported in tumour cells and suggested as a possible target for chemotherapeutic drugs. The underlying defect has not been extensively researched, nor have levels of sulphur amino acids been examined in these cells. This study compared two rat liver tumour cell lines. One was found to be methionine dependent (HTC) and the other found to be methionine independent (Phi-1). The methionine-dependent cell line (HTC) was discovered to contain markedly less methionine synthase activity, the enzyme activity being less responsive to methionine concentration than in the methionine-independent cells (Phi-1). HTC cells had lower cysteine requirements and contained larger concentrations of reduced glutathione (GSH) and taurine than the Phi-1 cells. Also, in contrast to Phi-1 cells, no glutathione was found in the media of the HTC cells, although large quantities of cysteinylglycine were detected. These results suggested that differences in methionine synthase activity might be partly responsible for methionine dependence and that methionine-dependent cells may have different metabolic requirements for other sulphur amino acids.

The correlation between urinary and liver taurine levels and between pre-dose urinary taurine and liver damage

Analysis of data from several studies has shown that urinary taurine levels are highly significantly correlated with liver taurine concentration in control rats. Furthermore, urinary taurine levels measured before dosing with various hepatotoxic agents are significantly correlated with serum AST and ALT values measured after dosing with hepatotoxicants. That is, animals with low urinary taurine values and therefore low liver taurine concentrations tend to show greater hepatic damage. These data suggest that taurine may have a protective function in the liver.