ABSTRACT
Title
Effects of Lisosan G on antioxidant and xenobiotic-metabolising enzymes in primary rat hepatocytes cultures
Authors
M. La Marca
Doctorate School in Cellular and Molecular Physiology, Pharmacology and Toxicology
Section of Pharmacology and Molecular Toxicology (2nd year)– University of Siena, Italy
Institute of Agricultural Biotechnology, CNR, via Moruzzi 1, 56100 Pisa, Italy
Doctorate School in Cellular and Molecular Physiology, Pharmacology and Toxicology
Section of Pharmacology and Molecular Toxicology (2nd year)– University of Siena, Italy
Institute of Agricultural Biotechnology, CNR, via Moruzzi 1, 56100 Pisa, Italy
Abstract
In recent years several plants, including grain extracts, exhibit antioxidant properties that could be useful in the prevention of oxidative stress reactions, such as those mediated by the formation of free radical species in different pathological situations.It has been recently shown that Lisosan G, a powder of grain of Triticum Sativum registered to the Italian Minister of Health as an integrator, did not interfer with cytochrome P450 and phase 2 enzymes, had an antioxidative properties and protective effects versus the carbon-tetrachloride-induced hepatotoxicity in rats (Longo 2007). In 2011, Longo et al. has also shown a protective effect in vivo of Lisosan G against the cisplatin induced toxicity. The authors suggested that the protective effect of Lisosan G could be associated mainly with the attenuation of the oxidative stress and the preservation in antioxidant enzymes.
In the present study, by using primary rat hepatocytes, we have investigated the effects of Lisosan G on the antioxidant and drug-metabolising enzymes at transcriptional, catalytic and immunoblotting levels. The isolation of hepatocytes was made by De Smet method (De Smet et al. 1998) using two layers of rat-tail collagene (type 1). Cells were treated for 24 hours with Lisosan G at the dose of 0,7 mg/ml. Then, hepatocytes were analyzed by PCR for the expression of NAD(P)H: quinone oxidoreductase-1 and heme oxygenase-1 genes. On the other hand, in the hepatocyte microsomes and 100000 x g supernatants, it was investigated the enzymatic activity of NAD(P)H: quinone oxidoreductase-1, glutathione-S-transferase, catalase, heme oxygenase-1, lipid peroxidation.Lisosan G was able to activate the transcription of NAD(P)H: quinone oxidoreductase-1, heme oxygenase-1 genes and the corresponding activities. In addition, it was also observed a significant induction of glutathione-S-transferase, catalase antioxidant activities. By western blotting analysis it was ascertained that heme oxygenase-1 levels protein was increased after the treatment. In principle, we observed an induction of antioxidant and xenobiotic-metabolising enzymes after treatment with Lisosan G.
In the present study, by using primary rat hepatocytes, we have investigated the effects of Lisosan G on the antioxidant and drug-metabolising enzymes at transcriptional, catalytic and immunoblotting levels. The isolation of hepatocytes was made by De Smet method (De Smet et al. 1998) using two layers of rat-tail collagene (type 1). Cells were treated for 24 hours with Lisosan G at the dose of 0,7 mg/ml. Then, hepatocytes were analyzed by PCR for the expression of NAD(P)H: quinone oxidoreductase-1 and heme oxygenase-1 genes. On the other hand, in the hepatocyte microsomes and 100000 x g supernatants, it was investigated the enzymatic activity of NAD(P)H: quinone oxidoreductase-1, glutathione-S-transferase, catalase, heme oxygenase-1, lipid peroxidation.Lisosan G was able to activate the transcription of NAD(P)H: quinone oxidoreductase-1, heme oxygenase-1 genes and the corresponding activities. In addition, it was also observed a significant induction of glutathione-S-transferase, catalase antioxidant activities. By western blotting analysis it was ascertained that heme oxygenase-1 levels protein was increased after the treatment. In principle, we observed an induction of antioxidant and xenobiotic-metabolising enzymes after treatment with Lisosan G.