Protective effect of Propolfenol® on induced oxidative stress in human spermatozoa.

The propolis extract was shown to possess the capacity to protect sperm membrane from the deleterious action of oxidative attack. Oxidative stress can induce propagation of a lipid peroxidation (LPO) chain reaction because spermatozoa contain high concentration of unsaturated fatty acids. This study aimed at evaluating in vitro the possible toxicity and/or the antioxidant properties of Propolfenol® in ejaculated human spermatozoa. A colorimetric assay determined the total flavonoid content by spectrophotometry and a high-performance liquid chromatography-diode array detection analysis the quantity of galangin, pinocembrin and caffeic acid phenylethilic ester (CAPE). Sperm parameters such as motility, vitality and DNA integrity were assessed utilising optical microscopy. The antioxidant properties Propolfenol® against LPO induced by tert-Butyl Hydroperoxide were evaluated using the C11-BODIPY581/591 probe. Chemical analysis of Propolfenol® revealed low quantities of galangin, pinocembrin and CAPE; cyclic voltammetry experiments showed that Propolfenol® may exert an antioxidant activity. A protective action of Propolfenol® (20 and 100 µg/ml) on induced LPO in human spermatozoa was detected. Propolfenol® may be proposed as the supplement in media for sperm preparation techniques or cryopreservation to counteract the increased presence of reactive oxygen species generated by these methods.

Evaluation of enzymatic and non-enzymatic antioxidants in seminal plasma of men with genitourinary infections, varicocele and idiopathic infertility.

UNLABELLED: This study was aimed to assess the antioxidant enzymatic and non-enzymatic compounds in semen of infertile men. Seventy-four infertile patients were grouped according to their clinical diagnosis: genitourinary infection, varicocele, idiopathic infertility. Semen samples of fertile men represent the control. Semen characteristics were evaluated by light and transmission electron microscopy (TEM). TEM data was quantified with a mathematical formula, which provides numerical scores. Spectrophotometric and HPLC methods were used to measure the amount of reduced (GSH), oxidised glutathione (GSSG), ascorbic acid (AA) and malondialdehyde (MDA, marker of lipid peroxidation) and the activity of glutathione reductase, catalase (CAT), glutathione peroxidase. Infertile groups showed significantly decreased values of sperm parameters vs. CONTROLS: In infection and varicocele groups, the seminal MDA levels were significantly increased when compared to controls (p < 0.001), indicating an alteration of oxidative status and a peroxidative damage. In infection and varicocele groups, AA levels were reduced (p < 0.05) vs. control; in the varicocele group, the GSH levels were also decreased (p < 0.05). Significantly higher CAT activity was observed in infection and varicocele groups vs. fertile men (p < 0.001 and p < 0.05 respectively). The GSH/GSSG ratio was significantly decreased in varicocele and idiopathic infertility groups vs. control (p < 0.01). The study of the alteration of a single parameter of oxidative stress or of the antioxidant system may not have a relevant clinical value to estimate male fertilising potential and the background of infertility causes, since complex and multifactorial mechanisms are involved in different pathologies. In our study, each pathology is characterised by a definite pattern of markers such as MDA and enzymatic and non-enzymatic antioxidant compounds. In the different pathologies related to infertility, the identification of the complex of involved parameters could be useful in the diagnosis, prognosis and in the choice of a possible treatment such as specific antioxidant supplements.

Thermal water of Vetriolo, Trentino, inhibits the negative effect of interleukin-1β on nitric oxide production and apoptosis in human osteoarthritic chondrocyte.

The thermal water of Vetriolo in Trentino, Italy (VW) has been known over 150 years for its therapeutic properties in the treatment of osteoarthritis (OA). This is a highly mineralized water, strongly acidic sulfate, rich in calcium, magnesium and iron and used for balneotherapy after dilution. The aim of our study was to investigate the possible in vitro effects of the VW in human OA chondrocytes cultivated in the presence or in the absence of Interleukin-1 beta (IL-1beta). OA chondrocytes were cultivated in Deionized Water (DW) (DW-DMEM, controls), or in one of three different VW-DMEM media, in which DW had been totally (100 percent) or in part (25 or 50 percent) substituted with VW. All samples were analyzed before and after treatment with IL-1beta at a concentration of 5 ng/ml. After 48 h, we evaluated the cell viability, the release of nitric oxide (NO) in culture medium, the inducible nitric oxide synthase (iNOS) expression, and the percentage of apoptosis and necrosis. Finally, we carried out a morphological assessment using a transmission electron microscope (TEM). Our data showed that VW alone at 25 or 50 percent concentration did not affect the viability of cultured OA chondrocytes, and determined a significant survival recovery rate in cultures stimulated with IL-1beta. On the contrary, the VW alone at 100 percent of concentration reduced, in a significant (P less than 0.05) manner, the cells viability. NO levels were low both in DW-DMEM cultures and in those reconstituted with 25 or 50 percent of VW, and were significantly (P less than 0.05) increased in cultures with 100 percent of VW. VW at 25 or 50 percent concentration significantly (P less than 0.001) reduced the NO production induced by IL-1beta. The data of the NO levels were confirmed by the immunocytochemistry assay for iNOS. Our experiments confirmed the pro-apoptotic effect of IL-1beta and demonstrated a protective effect of VW at 25 or 50 percent concentration. These findings were confirmed by TEM. In conclusion, our study demonstrated that VW alone at 25 or 50 percent concentration modifies neither morphology nor NO production and neither iNOS expression nor apoptosis, but it inhibits the negative effects of IL-1beta in chondrocytes cultures.

Distribution of alpha-, gamma (+beta)- and delta-tocopherol in the seminal plasma, spermatozoa and seminal vesicles of rabbit.

Dietary vitamin E supplementation plays a key role in animal reproduction by protecting germ cells from oxidative damage. Recently, alpha-tocopherol homologues (namely, beta-, gamma- and delta-tocopherol) have been the object of increasing research because of their peculiar nonantioxidant properties. We found that these tocol-derived compounds are not homogeneously distributed among semen components. Alpha-T was the major vitamin E homologue found in all semen fractions. Half of the total gamma (+beta)-T was found in germ cells, while more than 50% of total delta-T was preferentially accumulated in seminal plasma. The concentration of various tocol-derived compounds depended on their relative amounts in diet and the competition for saturable enzymes implicated in their metabolism. A higher concentration of delta-T in seminal plasma may be related to its more polar nature. However, the biological function of this compound in semen remains to be cleared. To our knowledge, this is the first study aimed at identifying alpha-tocopherol homologues in rabbit semen fractions.

Evaluation of the antifertility activity of the broom Spartium junceum in the mammalian male.

Male adult rabbits and rats treated with Spartium junceum showed a significant decrease in fertility, demonstrated by a lower number of pregnancies. The target of the drug seems to be the acrosomal protease system, the activity of which appears greatly reduced, while the morphology of testicular cells and epididymal spermatozoa is only partially affected. The antifertility effect is completely reversible.