A carnosine analog with therapeutic potentials in the treatment of disorders related to oxidative stress.

Interactive relationships among metabolism, mitochondrial dysfunction and inflammation at skeletal muscle level play a key role in the pathogenesis of disorders related to oxidative stress. Mitochondrial dysfunction and oxidative stress result in cellular energy deficiency, inflammation and cell death inducing a vicious cycle that promotes muscle wasting. The histidine-containing dipeptides, carnosine and anserine, are carbonyl scavengers whose cytoprotective contributions extend beyond the antioxidant defence, but the physiological meaning of these capacities is actually limited. In the present study, we compared and investigated the potential protective effects of three different histidine-containing dipeptides: carnosine, anserine and carnosinol, a carnosine-mimetic new compound, against oxidative stress induction in rat L6 skeletal muscle cells. The hydrogen peroxide induced-oxidative stress significantly altered cell morphology, induced apoptosis, oxidative stress and inflammation, decreased mitochondrial peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α)/sirtuin3 pathway and the antioxidant system. Notably, all three investigated dipeptides in the present study, with a different extent and in a concentration-dependent manner, reduced myotube oxidative stress, apoptosis and inflammation. The present study underlined that carnosinol, maintaining the safety condition of carnosine and anserine, was the more efficient studied dipeptide in the preservation of mitochondrial environment mediated by PGC-1α and sirtuin3 expression and thereby in the reduction of oxidative stress-related alterations in this in vitro skeletal muscle model. Furthermore, we observed that carnosinol's antioxidant effects are not blocked inhibiting sirtuin3, but are maintained with almost the same extend, indicating its multiple capacities of reactive carbonyl species-scavenging and of mitochondrial modulation through PGC-1α. In conclusion, carnosinol retained and surpassed the efficacy of the well-known investigated histidine-containing dipeptides improving oxidative stress, inflammation and also cell metabolism and so becoming a greatly promising therapeutic carnosine derivate.

Obesity-related dysfunction of the aorta and prevention by melatonin treatment in ob/ob mice.

In this study, we hypothesized that melatonin administration can minimize alterations in aorta morphology in an animal model of obesity (ob/ob mice). The animals were divided into four groups: (i) control lean mice, (ii) control lean mice treated with melatonin, (iii) ob/ob mice and (iv) ob/ob mice treated with melatonin. The synthetic melatonin was dissolved in 1% ethanol and added to the drinking water from postnatal week 5-13 at a final dose of 100 mg/kg body weight/day. Compared with the obese mice, melatonin intake was associated with a significant decrease in body weight and water consumption. Histological analysis showed that the aortic wall of ob/ob mice had a high Tunica media/lumen ratio and that the elastic fibers in the media layer appeared disrupted and degraded. Moreover, the aorta of ob/ob mice displayed a higher degree of collagen accumulation in the Tunica media compared to the normal aorta. The aorta of ob/ob mice treated with melatonin had a lower Tunica media/lumen ratio and collagen accumulation in comparison with untreated ob/ob mice. Our results showed that whereas melatonin had no apparent histological effects on the aorta in lean mice with normal weight, its administration in ob/ob mice can lead to a reduction in body weight and can ameliorate aorta histopathological dysfunction. This experimental study indicates an apparent protective role for melatonin on the aorta in obesity and melatonin could possibly be an effective tool in the management of obesity-related vascular complications.

Vascular endothelial cells and dysfunctions: role of melatonin.

Several pathological conditions, including hypertension, atherosclerosis, diabetes, ischemia/reperfusion injury and nicotine-induced vasculopathy, are associated with vascular endothelial dysfunction characterized by altered secretory output of endothelial cells. Therefore there is a search for molecules and interventions that could restore endothelial function, in particular augmenting NO production, reducing the generation of free radicals and vasoconstrictors and preventing undesired inflammation. The pineal hormone melatonin exhibits several endothelium protective properties: it scavenges free radicals, activates antioxidant defence enzymes, normalizes lipid and blood pressure profile and increases NO bioavailability. Melatonin improved vascular function in experimental hypertension, reducing intimal infiltration and restoring NO production. Melatonin improved the NO pathway also in animal models for the study of diabetes and prevented NO down-regulation and adhesive molecules up-regulation in nicotine-induced vasculopathy. The protection against endothelial damage, vasoconstriction, platelet aggregation and leukocyte infiltration might contribute to the beneficial effects against ischemia-reperfusion injury by melatonin. Therefore, melatonin administration has endothelium-protective potential in several pathological conditions. Nevertheless, it still needs to be established, whether melatonin is able to revert already established endothelial dysfunction in these conditions.

Peripheral purinergic receptor modulation influences the trigeminal ganglia nitroxidergic system in an experimental murine model of inflammatory orofacial pain.

ATP plays an important role as an endogenous pain mediator generating and/or modulating pain signaling from the periphery to the central nervous system. The aim of this study was to analyze the role of peripheral purinergic receptors in modulation of the nitroxidergic system at a trigeminal ganglia level by monitoring changes in nitric oxide synthase isoforms. We also evaluated Fos-positive neurons in brainstem (spinal trigeminal nucleus) and pain-related behavior. We found that local administration of the P2 purinergic receptor antagonist pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS) decreased face-rubbing activity, nitric oxide synthase isoform expression in trigeminal ganglia, and Fos expression in spinal trigeminal nucleus after subcutaneous injection of formalin. These results suggest a role for peripheral P2 purinergic receptors in orofacial pain transmission through modulation of the nitroxidergic system. .

MAK-5 treatment enhances the nerve growth factor-mediated neurite outgrowth in PC12 cells.

The effects of an ayurvedic compound (MAK-5) alone or together with nerve growth factor (NGF) on the neurite outgrowth of PC12 cells was studied. PC12 cells treated with NGF alone showed a clear neurite outgrowth with a decrease of the proliferation at the dose higher than 5 ng/ml. MAK-5 alone does not induce significant neurite outgrowth in the PC12 cells and does not decrease the proliferation. The PC12 cells treated with NGF supplemented with MAK-5 showed a well-evident morphological differentiation also at low doses of NGF (less than 5 ng/ml), however, the proliferation does not decrease. We suggest that MAK-5 could contain some differentiating agents that are able to potentiate NGF inducing neuronal differentiation in PC12 cells without decreasing the cell proliferation.

Cyclosporine-A treatment inhibits the expression of metabotropic glutamate receptors in rat thymus.

We have evaluated the expression of metabotropic glutamate receptors (mGluR subtypes 2/3, 4 and 5) in rat thymus under normal and experimental conditions after 2 and 21 days of cyclosporine-A treatment. In normal rats, immunohistochemical analysis showed that expression of mGluRs was high in dendritic cells and lymphocytes of the medulla whereas it was weak in lymphocytes of the cortex. However, there were some differences in the expression of mGluRs subtypes. mGluR5 showed strong expression in lymphocytes of medulla and dendritic cells. mGluR2/3 and mGluR4 were moderately expressed in lymphocytes and dendritic cells of the medulla and weakly in cortical lymphocytes. Immunoblotting showed moderate levels of mGluR2/3 and mGluR4 and strong levels of mGluRS. After 2 days of cyclosporine-A treatment, we observed by immunohistochemistry and immunoblotting a distinct decrease in all mGluRs and their expression had almost completely disappeared after 21 days of treatment. The results clearly indicate that: 1) mGluR2/3, 4 and 5 are widely expressed in thymic cells; 2) the mGluR5 subtype is expressed most strongly in medullary cells; and 3) cyclosporine-A rapidly inhibits expression of all mGluR subtypes after 2 days of treatment and their complete disappearance after prolonged treatment. These findings may indicate a possible mechanism by which cyclosporine-A produces its immunosupressive effects.