l-carnitine protects rat hepatocytes from oxidative stress induced by T-2 toxin.

CONTEXT: Oxidative stress and mitochondrial dysfunction are thought to be the main mechanism of T-2 toxin toxicity. T-2 toxin is the most potent trichothecene mycotoxin which is present in agricultural products. L-carnitine, besides its anti-oxidative properties, facilitates the transportation of long-chain fatty acids in to mitochondrial matrix. OBJECTIVE: In this study we tested whether L-carnitine, an antioxidant and a facilitator for long-chain fatty acid transportation across mitochondrial membranes, could protect rat hepatocytes against toxicity induced by T-2 toxin. MATERIALS AND METHODS: L-carnitine in low and high doses (50 and 500 mg/kg) was administered for five consecutive days to male Wistar rats. Hepatocytes were isolated and freshly exposed to appropriate concentration of T-2 toxin for 2 h followed by oxidative stress and cell death evaluations. RESULTS: Glutathione depletion, ROS overproduction and mitochondrial membrane potential collapse were determined under T-2 toxin exposure. Pretreatment with L-carnitine particularly at high-dose reduced toxicity and prevented the hepatocytes from abnormal caspase-3 activity and apoptosis. CONCLUSION: Low toxicity of L-carnitine and its mitochondrial protective effects promises an effective way to reduce or prevent the toxicity induced by certain environmental pollutants, including T-2 toxin.

Determination of the mutagenicity potential of dillsun herbal medicine by single cell gel electrophoresis in rat hepatocytes.

BACKGROUND: Traditional medicines are among the oldest medicines and their extensive use in the recent years reflects the public's interest in alternatives to conventional medicine. OBJECTIVES: The aim of this study was to investigate the genotoxicity of Dillsun herbal medicine in DNA damage of rat hepatocytes compared to sodium dichromate using a comet assay technique. MATERIALS AND METHODS: Male Wistar rats were caught and their liver was washed with a perfusion buffer, followed by another wash with collagenase buffer. Hepatocytes were isolated and transferred on to a petri dish which contained a washing buffer. Hepatocytes were then separated and the cells were filtered and centrifuged at 1500 rpm for 3 minutes. The hepatocytes were counted using neubauer slides and kept in a bioreactor for 30 minutes. Cells were then exposed to different doses of Dillsun such 0.2, 1, 2.5, 5 and 10 mg/mL. Sodium dichromate was the positive control and incubated buffer was used as a negative control. Cell suspensions were placed on slides pre-coated with low melting point agarose and were covered with agarose gel. Agarose gels were then lysed and electrophoresis was done, followed by neutralization and staining. Slides were analyzed by fluorescence microscopy. The size and extent of DNA damage visualized by this technique was evaluated by examining cells. Migration behavior was classified according to the Kobayashi pattern. RESULTS: The results indicated that with an increase of Dillsun dose, the mutagenicity index slightly increased but compared to the positive control, there were significant differences, which suggests that the crude extract of Dillsun in vitro did not have mutagenic effects. CONCLUSIONS: In conclusion the results showed that Dillsun has no mutagenic effects when compared to the positive control. Although by increasing the Dillsun dose, DNA damage also increased but this increase was not significant.

Determination of the Mutagenicity Potential of Sankol Herbal Medicine by Single Cell Gel Electrophoresis in Rat Hepatocytes in Comparison With H2O2.

BACKGROUND: The increasing use of herbal drugs and their ease of accessibility and availability have necessitated the use of mutagenicity test to analyze their toxicity and safety. OBJECTIVES: This study aimed to evaluate the genotoxicity of Sankol herbal medicine in DNA breakage of rat hepatocytes in comparison with H2O2 by single cell gel electrophoresis technique or comet assay. MATERIALS AND METHODS: In the current study hepatocytes were prepared from male wistar rats. Hepatocytes cells were counted and kept in a bioreactor for 30 minutes,then cells were exposed to Sankol herbal medicine at doses of 100, 200 and 400 µl/ml. Buffer 4 (incubation buffer) and H2O2 were used for one hour as negative and positive control respectively. After 30 minutes cell suspension with low melting point agarose was put on precoated slides and covered with agarose gel. Then lysing, electrophoresis, neutralization and staining were carried out. Finally the slides were analyzed by fluorescence microscope. The parameter under this analysis was the type of migration which was determined according to Kobayashi pattern. RESULTS: Results of the study indicated that by increasing the dose of Sankol herbal medicine, the DNA damage slightly increased (P < 0001). CONCLUSIONS: In overall compared to the positive control, significant differences were observed which indicated that the crude extract of Sankol in vitro did not have mutagenic effect.

Determination of the mutagenicity potential of supermint herbal medicine by single cell gel electrophoresis in rat hepatocytes.

PURPOSE: The increasing use of herbal drugs and their easy availability have necessitated the use of mutagenicity test to analyze their toxicity and safety. The aim of this study was to evaluate the genotoxicity of Supermint herbal medicine in DNA breakage of rat hepatocytes in comparison with sodium dichromate by single cell gel electrophoresis technique or comet assay. METHODS: Hepatocytes were prepared from male wistar rats and were counted and kept in a bioreactor for 30 minutes. Then cells were exposed to the Supermint herbal medicine at doses of 125, 250 and 500 µl/ml. Buffer 4 (incubation buffer) and sodium dichromate were used as negative and positive control for one hour respectively. Then cell suspension with low melting point agarose were put on precoated slides and covered with agarose gel. Then lysing, electrophoresis, neutralization and staining were carried out. Finally the slides were analyzed with fluorescence microscope. The parameter under this analysis was the type of migration which was determined according to Kobayashi pattern. RESULTS: With increased dose of Supermint herbal medicine the DNA damage was slightly increased (P<0001). Conlusion: In overall compared to the positive control significant differences is observed which convinced that the crude extract of Supermint in vitro did not have mutagenic effect. Conlusion: In overall compared to the positive control significant differences is observed which convinced that the crude extract of Supermint in vitro did not have mutagenic effect.