Francesco Vizzarri

Ocular tissues and fluids oxidative stress in hares fed on verbascoside supplement

Abstract The influence of a prolonged diet supplemented with the powerful antioxidant verbascoside on the oxidative state of 20 healthy hares eye fluids and tissues has been studied. Verbascoside was dosed at 2, 3, 4 mg/die and the impact on the oxidative state of ocular tissues and fluids was tested by TBARS (thio barbituric acid reactive substances) and TEAC (trolox equivalent antioxidant capacity) assays. The percentage of change in antioxidant activity increased largely in retina and lenses at a daily verbascoside dose of 3 mg, whereas for optic nerve and vitreous humor the higher antioxidant capacity was measured at 4 mg/die verbascoside dose. The present findings demonstrate that verbascoside supplementation is able to protect ocular tissue and fluids from naturally occurring oxidation and that its protective effect depends on the daily dose, being maximum up to 3 mg/die.

Effects of verbascoside-based diet on blood and plasma constituents of rabbits

Objective: Oxidative stress brought on by free radicals can lead to an increased risk of some chronic pathologies. Antioxidants can scavenge free radicals by turning them into nonradical and nontoxic metabolites. The purpose of this study is to investigate the effect of a phenylpropanoid glycosides–based prolonged diet on blood constituents in animals. Methods: Tests were carried out on healthy New Zealand white rabbits and the following parameters were evaluated at baseline and after 90 days’ follow-up: plasma triglycerides, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, aspartate transaminase, alanine aminotransferase, bilirubin, the reactive oxygen metabolites, thiobarbituric acid–reactive substances, vitamin A, and vitamin E. The same parameters were analyzed in an age- and sex-matched animal control group. Results: We first defined the concept of average rate and then used it to calculate, by experimental data fitting, the formation or destruction rate of some blood or plasma constituents as a function of the daily dose. The results indicate that the effects can be categorized into 2 classes. The first includes the effects that produce monotonously continuous changes with daily dose, and the second includes those that exhibit a saturating trend. Conclusions: The experimental results suggest that high doses of verbascoside can potentially cause adverse effects through prooxidative effects. Risk is increased by the use of pharmacological doses of polyphenols in prevention, treatment, and as dietary supplements.

The effects of fermented wheat powder (Lisosan G) on the blood lipids and oxidative status of healthy rabbits

A number of studies have shown that whole grain products have strong antioxidant effects. Lisosan G, a powder produced from the fermentation of Triticum aestivum, has shown hepatoprotective effects. The aim of this paper was to evaluate the effects of Lisosan G on the serum lipid profile and oxidative status of healthy rabbits. Forty rabbits were fed either a normal diet or a diet containing 10 g, 20 g or 30 g Lisosan G/kg of feed for 60 days. The supplementation of Lisosan G did not cause any alteration of the aspartate aminotransferase (AST), alanine aminotransferase (ALT), bilirubin or creatinine concentrations, but caused a reduction in triglycerides, in low density lipoprotein (LDL) cholesterol, and an increase in high density lipoprotein (HDL) cholesterol. The antioxidant activity of Lisosan G has been confirmed on the basis of the reduction in the blood reactive oxygen metabolites (ROMs) and in the level of the thiobarbituric acid reactive substances (TBARS) together with increased vitamin A and E concentrations in the blood. Moreover, Lisosan G also caused the induction of antioxidant enzymes in the liver and kidney of the treated rabbits. This study has shown the ability of Lisosan G to improve the lipid profile and antioxidant status of the blood in healthy rabbits.

Corneal Epithelial Wound Healing Promoted by Verbascoside-Based Liposomal Eyedrops

Different liposomal formulations were prepared to identify those capable of forming eyedrops for corneal diseases. Liposomes with neutral or slightly positive surface charge interact very well with the cornea. Then these formulations were loaded with verbascoside to heal a burn of corneal epithelium induced by alkali. The cornea surface affected involved in wound was monitored as a function of time. Experimental results were modeled by balance equation between the rate of healing, due to the flow of phenylpropanoid, and growth of the wound. The results indicate a latency time of only three hours and furthermore the corneal epithelium heals in 48 hours. Thus, the topical administration of verbascoside appears to reduce the action time of cells, as verified by histochemical and immunofluorescence assays.

Cytogenetic tests reveal no toxicity in lymphocytes of rabbit (Oryctolagus cuniculus, 2n=44) feed in presence of verbascoside and/or lycopene

Phenylpropanoid glycosides (PPG), like other phenolic compounds, are a powerful antioxidants and the Verbascoside (VB) is one of the most active of them. A previous study, by using in vitro exposure of blood human lymphocytes to Verbascoside, reported a significant increasings of chromosome fragility compared to control. In the present study, four homogeneous groups of rabbits were used to test in vivo the VB and/or Lycopene (LP) by feeding the animals without VB and LP (control), in presence of VB or/and LP for 80 days. Lymphocyte cell cultures were performed in three different times: 0, 40 and 80 days of the experiment and the cytogenetic tests that we used [CA-test (Chromosome Abnormalities in terms of chromosome and chromatid breaks) and Sister Chromatid Exchange (SCE-test)] have revealed no mutagenic effects on chromosomes. Indeed, mean values/cell of CA and SCE decreased during the experiment with some difference among and within groups, with significant decreasing value only for some group. The study shows clear evidence that diets rich in Verbascoside (and/or Lycopene) do not originate any mutagenic activity, resulting no cytotoxic for the animals and, suggesting a possible their use in both animal and human diets.

The effect of water-restriction on various physiological variables in intensively reared Lacaune ewes

The effects of water restriction on rectal temperature, respiratory rate and selected blood variables in intensively reared Lacaune ewes were evaluated. The tests were carried out over the course of 28 days in spring on 30 lactating sheep divided into three groups of 10 animals each, matched by lactation status and body weight and allocated into individual boxes. The animal groups were as follows: control group W100 with drinking water for the whole day (24 h/24 h), while the two experimental groups each received 80% (W80 group) and 60% (W60 group) of the water ration of the control group, respectively. The feed, in according with physiological and production needs of the animals, was administered in three daily meals (08:00, 14:00 and 20:00 h). Water intake was recorded three times per day (at 08:00, 14:00 and 20:00 h) and the daily feed intake at 08:00 h, while the rectal temperature, respiratory rate were measured and blood samplings were carried out on Day 0, Day 14 and Day 28. In water-restriction groups (W80 and W60) a lower (P < 0.01) feed intake of meadow hay, and a lower ingestion of alfalfa hay pellet in the W60 group compared to W100 group were observed. In addition, the water restriction regimen resulted in a significant decrease of respiratory rate (P < 0.05) and an increase in serum total protein, urea, creatinine, sodium, chlorine, reactive oxygen metabolites, cortisol, haemoglobin and mean corpuscular haemoglobin concentration (P < 0.05), red blood cell counts, and haematocrit (P < 0.01). These results show the important role of water, as limiting factor for animal breeding in low-water availability environments. Where possible, the management of low-dairy breeds should be well adapted to support the productive performance.

Polyphenolic characterisation of plant mixture (Lisosan® Reduction) and its hypocholesterolaemic effect in high fat diet-fed mice

Abstract Lisosan® Reduction is a plant mixture produced from powder of fermented Triticum aestivum (Lisosan® G), Desmodium adscendens, Malus domestica, Picrorhiza kurroa and Hordeum vulgare. The aim of this study was to characterise the phenolic profile of Lisosan® Reduction and to evaluate the effects of aqueous extract on mice fed a high fat diet (HFD). Syringic acid, trans sinapic acid and neochlorogenic acid were identified by HPLC-DAD to be the dominant polyphenols of Lisosan® Reduction, followed by vitexin, trans p-coumeric acid and trans ferulic acid. Mice treated with aqueous extract of Lisosan® Reduction (60 mg/kg b.w.) showed a significant decrease of serum cholesterol, glucose and triglycerides level and a significant increase of CYP7A1 gene expression, compared to HFD group.

Genotoxicity and cytotoxicity of Aloysia polystachia: an in vivo study in rabbits

In the search of new compounds useful for the treatment of neurological disorders, medicinal plant research has progressed widely in the last decade, particularly because of the hypothesis that natural extracts possess a low or absent toxicity. Aloysia polystachia (Griseb.) Moldenke belonging to the family of Verbenaceae has been used in the traditional medicine for a variety of indications and as a sedative. Recently, some evidence reported that it exhibits some antidepressant- and anxiolytic-like effects involving the modulation of GABA system in preclinical studies in mice (1). However no investigations have been performed on its (geno)toxicity in long-term studies as it would be required for its use in neurological pathologies. In this study, we investigated the genotoxic potential of a water soluble extract of Aloysia polystachia leaves, administered orally every day for 90 days to 20 New Zealand white rabbits homogeneous for weight and age. Subjects were divided into four groups: the control group, and three experimental groups fed with a diet supplemented with 1g/kg; 1.5 g/kg and 2g/kg of Aloysia Polystachia, then peripheral blood was drawn from the vein auricularis marginalis before and after 45 and 90 days of treatment. Cytogenetic analysis was performed on each subject at each time point and chromosome aberrations (structural and numerical) and mitotic index were used as a measure of DNA damage and cytotoxicity (2). Results indicated that Aloysia polystachia extract induces a significant increase in the percentage of aberrant cells as well as in the aberration frequency (mainly chromatid breaks and fragments) associated with a decreasing trend of the mitotic index. The DNA damage was particularly higher after the first 45 days of treatment whereas it remains significantly high but almost unvaried until the end of the treatment. These data suggest that Aloysia polystachia extract has genotoxic and cytotoxic activity, even though further investigations are required to assess which compound of the extract could be responsible for the observed effects.