Specific Activities Of Gamma-glutamyl Transpeptidase Research Articles

In vitro induction of differentiation by ginsenoside Rh2 in SMMC-7721 hepatocarcinoma cell line.

The aim of this study was to investigate the effects of ginsenoside Rh(2) (G-Rh(2)) on differentiation of SMMC-7721 hepatocarcinoma cell line in culture. We studied G-Rh(2)-induced differentiation of SMMC-7721 cells through cell proliferation, cell morphology, ultrastructure, cell cycle, cell function and metabolism. The proliferation of treated cells was inhibited, the morphology and ultrastructure seemed normal, the secretory amount and expression of alpha-foetoprotein, and the specific activity of gamma-glutamyl transpeptidase, and heat-resistant alkaline phosphatase were all significantly decreased, the secretory amount of albumin and alkaline phosphatase activity were remarkably increased, and the cell was arrested at the G(1)/G(0) phase. Furthermore, G-Rh(2) induced elevated expression of the cyclin-dependent kinase inhibitor p21(WAF1) and p16(INK4a), and declined expressions of cyclin D1 and cyclin E. In addition, G-Rh(2) almost completely inhibited telomerase activity, as measured by polymerase chain reaction-based telomeric repeat amplification protocol coupled with enzyme-linked immune sorbent assay, and human telomerase reverse transcriptase mRNA. Based on these data, it is suggested that G-Rh(2) could induce cell differentiation tending to normal and effectively reduce telomerase activity with affecting transcription levels of human telomerase reverse transcriptase, paralleling the induction of cell differentiation.

1,25-dihydroxyvitamin D3 regulates the synthesis of gamma-glutamyl transpeptidase and glutathione levels in rat primary astrocytes.

Astrocytes play a pivotal role in CNS detoxification pathways, where glutathione (GSH) is involved in the elimination of oxygen and nitrogen reactive species such as nitric oxide. We have previously demonstrated that the specific activity of gamma-glutamyl transpeptidase (gamma-GT), an enzyme of central significance in GSH metabolism, is regulated in vivo in astrocytes by 1,25-dihydroxyvitamin D3 (1,25-D3). The aim of the present work was to investigate, in primary cultures of newborn rat astrocytes, the effects of this hormone on gamma-GT synthesis and on GSH and nitrite levels after lipopolysaccharide (LPS) treatment. This study demonstrates that both gamma-GT gene expression and specific activity, induced by LPS, are potentiated by 1,25-D3. In contrast, 1,25-D3 does not regulate the expression of other enzymes involved in astrocyte detoxification processes, such as superoxide dismutase or GSH peroxidase. In parallel, 1,25-D3 enhanced intracellular GSH pools and significantly reduced nitrite production induced by LPS. Taken together, these results suggest that gamma-GT, GSH, and 1,25-D3 play a fundamental role in astrocyte detoxification pathways.

Sex differences in oxidative damage in ddY mouse kidney treated with a renal carcinogen, iron nitrilotriacetate

Iron-induced free radical injuries in male and female ddY mice, especially the sex difference and its mechanisms, were studied after an i.p. injection of a renal carcinogen, ferric nitrilotriacetate. Male mice were much more susceptible to iron-induced free radical injuries than female mice. Oxidative modification of proteins and DNA occurred more strongly in males than in females, as measured by protein carbonyl content and 8-hydroxydeoxyguanosine, respectively. Histochemical detection of 4-hydroxy-2-nonenal-modified proteins using an antibody and DNA fragmentation as detected by the TUNEL method also showed that males are more severely damaged than females, especially in the proximal convoluted tubules. These results could not be explained by the difference in iron status between male and female mice. In fact, the toxic so-called 'free' iron in serum and kidney were not different between male and female mice and storage iron, such as ferritin and hemosiderin, was also comparable in both kidneys. In previous studies we proposed the glutathione cycling hypothesis to explain the sex differences. The half-life of glutathione in the kidney was significantly shorter in males (29 min) than in females (57 min), as determined by the glutathione decrease after buthionine sulfoximine treatment, a specific inhibitor of glutathione synthesis. The specific activity of gamma-glutamyltranspeptidase (EC 2.3.2.2) in female mice was 73% of that in male mice. These results suggest that the faster glutathione turnover in males could account for the higher susceptibility to oxidative injury by supplying the reducing equivalent that reduces Fe(III) to Fe(II), thereby facilitating iron-catalyzed free radical reactions.

γ-Glutamyl-transpeptidase in lymphatic tissues ofMastomys natalensis during an infection withAcanthocheilonema viteae

During Acanthocheilonema viteae infection, the specific activity of gamma-glutamyltranspeptidase (gamma-GT) increased in peritoneal exudate cells and bone marrow and decreased in lymph nodes of Mastomys natalensis throughout the course of infection. However, though there was an increase in specific activity of gamma-GT in thymus and spleen during the prepatent phase of A. viteae infection, the level either returned to normal or decreased during the latent phase of infection. A close correlation was observed between the host's immune status during A. viteae infection and the level of gamma-GT in lymphoid tissues.

Enzyme profile of rat bile ductular epithelial cells in reference to the resistance phenotype in hepatocarcinogenesis

An extensive bile ductular cell hyperplasia with the formation of well-differentiated bile ductules is the most prominent feature of rat liver at 6 to 15 weeks after bile duct ligation. We have improved our previous cell isolation procedure and are now routinely able to obtain from such livers high yields of viable bile ductular epithelial cells. These cells were characterized with respect to their specific activities of gamma-glutamyl transpeptidase and beta-glucuronidase and of select Phase I and Phase II enzymes of biotransformation. At the time of their isolation, only a very small number of the bile ductular epithelial cells were observed to be in DNA synthesis. In addition, in histological sections prepared from intact hyperplastic bile ductular tissue isolates, only the bile ductular epithelial cells exhibited histochemical staining for gamma-glutamyl transpeptidase activity. Typically, greater than 95% of the cells isolated from this tissue were also found to be histochemically positive for gamma-glutamyl transpeptidase activity, and no hepatocytes were seen contaminating this cell population. Biochemically, the isolated bile ductular cells exhibited a gamma-glutamyl transpeptidase specific activity that was 100 times higher than that of hepatocytes isolated at the same time from the bile duct-ligated rats and more than 300 times higher than the specific activity of the enzyme of freshly isolated normal rat hepatocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

Cellular enzymatic differentiation in brain vessels of normal and protein-deprived rats. Biochemical studies.

Brain capillary development was studied in normal and protein-deprived rats using the specific activity of alkaline phosphatase (EC 3.1.3.1) and gamma-glutamyl transpeptidase (EC 2.3.2.1) in whole brain homogenates and microvessels obtained by gradient centrifugation according to Orlowski et al. (1974). Pre- and postnatal protein deprivation was induced by a 50% reduction in the dietary protein content. The density of microvessel fragments changed during development. Most of the early developmental increase in the specific activity of both enzymes in whole brain homogenates of normal rats can probably be explained by a rapid formation of new capillary segments. The increase in specific activity of gamma-glutamyl transpeptidase in microvessels was interpreted as a sign of cellular differentiation. Protein deprivation resulted in reduced specific activity of both enzymes in whole brain homogenates of 30-day-old rats, probably as a result of the decreased length per volume of the cerebral capillary network at this age (Conradi et al. 1979a). Signs of impaired endothelial growth were also present in the protein-deprived rats since the distribution of microvessel fragments in the 30-day-old protein-deprived rats was similar to that in 3-week-old normal rats. The specific activity of alkaline phosphatase was decreased in the microvessel fractions of 30- and 96-day-old protein-deprived rats, apparently signifying an effect of the protein deprivation on the endothelial cells. These effects of protein deprivation on the brain capillary endothelial transport system may have negative consequences for growth and function in the brain.

Glutathione-degrading enzymes of microvillus membranes.

Microvillus membranes from rat kidney, jejunum, and epididymis have been purified by the Ca precipitation method. The membranes exhibit enrichment in specific activities of gamma-glutamyl transpeptidase, aminopeptidase M, and a dipeptidase. The latter has been characterized and shown to be the principal activity responsible for the hydrolysis of S derivatives of Cys-Gly (including cystinyl-bis-glycine (Cys-bis-Gly) and 5-hydroxy-6-S-cysteinylglycyl-1-7,9-trans-11,14-cis-eicosatetraenoic acid (leukotriene D4)). A method is described for the simultaneous purification of papain-solubilized forms of the three enzymes from renal microvilli. Dipeptidase (Mr = 105,000) appears to be a zinc metalloprotein composed of two Mr = 50,000 subunits. The enzyme is severalfold more effective in the hydrolysis of dipeptides than aminopeptidase M. Dipeptidase, in contrast to aminopeptidase M, is inhibited by thiol compounds; Cys-Gly, in particular, is a potent inhibitor (Ki = 20 microM). The inhibition of dipeptidase by thiols has been employed to probe the relative significance of dipeptidase and aminopeptidase M in the metabolism of glutathione and its derivatives at the membrane surface.