Melatonin ameliorates H2O2-induced oxidative stress through modulation of Erk/Akt/NFkB pathway

BACKGROUND: Improper control on reactive oxygen species (ROS) elimination process and formation of free radicals causes tissue dysfunction. Pineal hormone melatonin is considered a potent regulator of such oxidative damage in different vertebrates. Aim of the current communication is to evaluate the levels of oxidative stress and ROS induced damage, and amelioration of oxidative status through melatonin induced activation of signaling pathways. Hepatocytes were isolated from adult Labeo rohita and exposed to H2O2 at three different doses (12.5, 25 and 50 µM) to observe peroxide induced damage in fish hepatocytes. Melatonin (25, 50 and 100 µg/ml) was administered against the highest dose of H2O2. Enzymatic and non-enzymatic antioxidants such as malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) was measured spectrophotometrically. Expression level of heat shock proteins (HSP70 and HSP90), HSPs-associated signaling molecules (Akt, ERK, cytosolic and nuclear NFkB), and melatonin receptor was also measured by western blotting analysis. RESULTS: H2O2 induced oxidative stress significantly altered (P < 0.05) MDA and GSH level, SOD and CAT activity, and up regulated HSP70 and HSP90 expression in carp hepatocytes. Signaling proteins exhibited differential modulation as revealed from their expression patterns in H2O2-exposed fish hepatocytes, in comparison with control hepatocytes. Melatonin treatment of H2O2-stressed fish hepatocytes restored basal cellular oxidative status in a dose dependent manner. Melatonin was observed to be inducer of signaling process by modulation of signaling molecules and melatonin receptor. CONCLUSIONS: The results suggest that exogenous melatonin at the concentration of 100 µg/ml is required to improve oxidative status of the H2O2-stressed fish hepatocytes. In H2O2 exposed hepatocytes, melatonin modulates expression of HSP70 and HSP90 that enable the hepatocytes to become stress tolerant and survive by altering the actions of ERK, Akt, cytosolic and nuclear NFkB in the signal transduction pathways. Study also confirms that melatonin could act through melatonin receptor coupled to ERK/Akt signaling pathways. This understanding of the mechanism by which melatonin regulates oxidative status in the stressed hepatocytes may initiate the development of novel strategies for hepatic disease therapy in future.

Efficacy of Basella alba and Tribulus terrestris extracts for production of monosex Nile tilapia, Oreochromis niloticus.

The present study was aimed to evaluate the efficacy of Basella alba and Tribulus terrestris for induction of masculinisation in Nile tilpia. B. alba leaves and T. terrestris seeds were extracted with water, ethanol, methanol, dichloromethane, hexane and successive methanol and mixed sex juveniles of Nile tilapia were subjected to dietary treatment with the extracts at the concentration of 0.5, 1.0 and 1.5 gm/kg feed. Treatment with both the plants showed no adverse effect on general fish health. There was no significant interaction effects (P>0.05) of solvent and concentration, and solvent and plant material for percentage of males. But, significant interaction effect (P<0.05) of concentration and plant material was observed for percentage of males. Also, there was significant interaction effect (P<0.05) of solvent, concentration and plant material for percentage of males. For dietary administration of B. alba leaves, the highest percentage of males (83.2±0.7) was obtained by treatment with ethanol extract at the concentration of 1.0 gm/kg feed. For all the solvents, the highest percentage of males was observed at the concentration of 1.0 gm/kg. But, in treatment with T. terrestris seeds, the highest percentage of males (88.9±1.1) was obtained with ethanol extract at the concentration of 1.5 gm/kg feed, which was also the highest percentage of males for all the treatment categories.

Effects of methyltestosterone, tamoxifen, genistein and Basella alba extract on masculinization of guppy (Poecilia reticulata).

The effects of different anti-estrogenic chemicals were evaluated in guppy, Poecilia reticulata on sex differentiation and survival. New-born fry of the fish were fed diets containing genistein (1 gm kg-1), tamoxifen (100 mg kg-1), methanol extract of Basella alba (1 gm kg-1) and 17α-methyltestosterone (60 mg kg-1) for 30 days. The treated groups showed no significant difference (P>0.05) in survival percentage while a significant increase (P<0.05) in percentage of males were observed in all the treatment groups compared to that in control. Control fish showed the lowest percentage (44.49%) of males while 17α-methyltestosterone treatment provided maximum (81.90%) males, followed by tamoxifen (80.09%), genistein (70.59%) and B. alba (63.55%). Intersex fish with male like colouration and gonopodium development, but female gonad structure was observed in all the treated groups. The nonsteroidal compounds evaluated in this study showed potential for affecting sex ratios although their potency was lower compared to 17α-methyltestosterone. Further studies are needed to determine an optimum treatment regime with these agents for induction of 100% sex reversal in guppy.