Incidences de la restriction calorique et du Nickel sur l’aromatase testiculaire du rat

This study was designed to determine whether intermittent fasting induces malnutrition that, according to many authors, accentuates the cytotoxic effects of environmental pollutants, or caloric restriction that reduces these effects. Ninety six male Wistar rats (180g) were divided into two groups: one group was fed daily (N) and the other group was fed every second day (J) for one month. At the end of one month, each group was then divided into two subgroups, one subgroup received an injection of 0.9% NaCI (groups NO and JO), the other subgroup received an injection of 4 mg/kg NiCI^b2 (groups NNi and JNi). Intermittent fasting was continued in parallel to treatment for 1, 3, 5 and 10 days. Under these experimental conditions, nickel increased testicular aromatase activity and altered total RNA, while no alteration of these biomarkers was observed with intermittent fasting. The combination of these two factors, nickel and intermittent fasting, did not amplify these effects. In contrast, protection of RNA by intermittent fasting was observed, especially overexpression of aromatase mRNA.     

Protective Role of Cactus Cladodes Extract on Sodium Dichromate-Induced Testicular Injury and Oxidative Stress in Rats

Cactus (Opuntia ficus-indica) is a xerophyte plant that belongs to the Cactaceae family. The present study was designed to investigate the possible protective effects of cactus cladodes extract (CCE) on sodium dichromate-induced testis damage in adult male Wistar rats. For this purpose, CCE at a dose of 100 mg/kg was orally administrated, followed by 10 mg/kg sodium dichromate (intraperitoneal injection). After 40 days of treatment, the rats were sacrificed, and the testes were excised for histological, lipid peroxidation (LPO), and antioxidant enzyme analyses. Sodium dichromate treatment significantly (P?<?0.01) decreased the body, testis, and accessory sex organ weights, sperm count and motility, and serum testosterone level. In addition, histological analysis revealed pronounced morphological alterations with tubular necrosis and reduction in the number of gametes in the lumen of the seminiferous tubules of sodium dichromate-intoxicated rats. Furthermore, exposure to sodium dichromate significantly (P?<?0.01) increased LPO level and decreased superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities in testis. Interestingly, pretreatment with CCE significantly (P?<?0.01) restored the serum testosterone level, sperm count, and motility to the levels of the control group. Moreover, CCE administration was capable of reducing the elevated level of LPO and significantly (P?<?0.01) increased SOD, CAT, and GPx activities in testis. Cactus cladodes supplementation minimized oxidative damage and reversed the impairment of spermatogenesis and testosterone production induced by sodium dichromate in the rat testis.     

Analyse morphométrique semi quantitative de l’histologie testiculaire au cours du vieillissement

Testicular aging is usually studied using sperm and quantitative hormone analysis. Testicular samples are obviously difficult to obtain from a control aging population. Body donations from the Anatomy Department of the Saint-Peres University provided access to testicular samples from deceased men between the ages of 53 to 102 years. We present the first results of a semiquantitative histological morphometric study of testicular aging. We studied a series of 39 subjects. After removal of the sample within the first 24 hours, several investigations were conducted. Macroscopic examination (volume, weight) was followed by histological examination and computer-assisted morphometric analysis: N.I.H images based on the following parameters: (i) transverse sections of the seminiferous tubules (total surface, thickness of the basal membrane, and nuclear density of Sertoli cells, spermatogonia, spermatocytes and spermatozoids; (ii) histological sections were studied for interstitial tissue, number of clusters and the surface occupied by Leydig cells (percentage per parenchyma area), their appearance, size and nuclear density were determined; (iii) this study was completed by visual count of the various cell types in the seminiferous epithelium. The results obtained on a series of 39 subjects aged from 53 to 102 showed various alterations, such as thickening of the tunica albuginea and basal membrane and intertubule hyalinization. The most frequent histological pattern of the aging testis is a mosaic of various seminiferous tubule lesions varying from tubules with complete although reduced spermatogenesis to entirely sclerosed tubules. Individual variations are extremely marked with major alterations of spermatogenesis as early as 60 years old, with atrophied Leydig cells and, on the contrary, preserved spermatogenesis until the age of 95 years.     

A combination of ascorbic acid and α-tocopherol or a combination of Mg and Zn are both able to reduce the adverse effects of lindane-poisoning on rat brain and liver

The purpose of this study, carried out on male Wistar rats, was to evaluate the beneficial effects of supplementation with ascorbic acid (Vit C) and α-tocopherol (Vit E) or with Mg and Zn upon lindane-induced damages in liver and brain. Under our experimental conditions, lindane poisoning (5 mg/kg body weight per day for 3 days) resulted in (1) an increased level of plasma glucose, cholesterol and triglycerides, (2) an increased activity of LDH, ALP, AST, ALT, (3) an oxidative stress in liver and brain as revealed by an increased level of lipids peroxidation (TBARS) and a decrease of glutathione-peroxidase, superoxide dismutase and catalase activities in liver and brain. In conclusion, both Vit C + E or Mg + Zn treatments display beneficial effects upon oxidative stress induced by lindane treatment in liver and brain.     

Influence du vieillessement sur la spermatogenèse: étude histologique et cytogénétique moléculaire au niveau testiculaire chez 46 sujets âgés de 29 à 102 ans

The harmful effect of maternal age on abnormal meiotic behaviour has been clearly established, but little is known about the effect of paternal age on chromosome malsegregations and the results of studies on this question are fairly controversial. The purpose of this study was to evaluate the influence of ageing on testicular histology and aneuploidy rate in testicular post-meiotic cells. A possible age-related risk has been suggested by the increased frequency of medical assisted reproduction techniques for older men. We analysed 36 testicular samples from subjects aged 61–102 years by histology and histomorphometry. We examined testicular cells with fluorescence in situ hybridisation (FISH). We studied six histological sections (20 cross-sectioned tubules) by computer-assisted morphometric analysis (Histolab). The study was based on the following parameters: for seminiferous tubules: surface area, thickness of the basement membrane, nucleus density (Sertoli cells, spermatogonia, spermatocytes and spermatozoids); for interstitial tissue: cluster number and surface area occupied by the Leydig cells, nucleus density. We analysed cells by FISH and a set of three probes X, Y and 18 (Abbot^o). The results were compared to those of a control group of testicular biopsies from 10 subjects (29–40 years) with obstructive azoospermia and normal histology. Results: The histomorphometric study showed various alterations including: thickening of the basement membrane when spermatogenesis was arrested. The number of germinal cells and the number of Sertoli cells decreased with increasing age and Leydig cell hypertrophy was observed with increasing age. Complete spermatogenesis was observed in men up to the age of 95 years old. The most sensitive step was pachytene. Spermatogonia can persist until the age of 98 years. The 36 elderly men were divided into 3 groups: preserved spermatogenesis (17 subjects; group 1), arrested spermatogenesis (4 subjects; group 2) and early disrupted spermatogenesis with only diploid cells or no cells (15 subjects; group 3). For the control group, post-meiotic cells (n=4,882) showed 50.35% X18, 48.55% Y18 and 1.1% of cells with aneuploidy. For elderly subjects with preserved spermatogenesis, post-meiotic cell analysis (n=4,738) showed 50.76% X18, 47.95% Y18 and 1.29% of cells with aneuploidy. Subjects with arrested spermatogenesis presented 47.96% X18, 37.75%Y18 and the aneuploidy rate among spermatids (n=98) was 14.28%. This rate was higher than those observed in controls and in group 1. In conclusion, we observed that spermatogenesis was possible until an advanced age (95 years). There appears to be an increased incidence of post-meiotic aneuploidy in the case of arrested spermatogenesis (14.28 vs 1.10%). The aneuploidy rate in the group of subjects with preserved spermatogenesis was not statistically different from that observed in the control group.     

Interactions d’une restriction calorique avec les effets du Nickel sur la reproduction chez le rat

Human epidemiological studies have demonstrated signs of a reduction, since 1960, of several parameters of the sperm count with an increase of certain male genital tract diseases. The increasing contamination of the environment by chemical compounds appears to be an aetiological factor. Various authors have also proposed the hypothesis that caloric restriction has a beneficial effect on health or longevity. This study was deisgned to compare the effects of nickel on the reproductive functions of rats fed either daily or every second day, in order to evaluate the possible beneficial effects of caloric restriction on rat fertility. This study was conducted with male and female Wistar rats, fed either daily (N), or every second day: intermittent fasting (F). After one month of this treatment, (N) and (F) rats were divided into 2 groups: one group received tap water (NO and FO groups), and the other received the same water enriched with nickel chloride (100 mg/L, NNi and FNi groups). Intermittent fasting was continued in parallel with nickel treatment with for 2, 4, 10, 16, 30 and 60 days. To study malonic dialdehyde (MDA) levels, nickel was administered by intraperitoneal injection at the dosage of 4 mg NiCl₂/kg of body weight for 1, 3, 5 and 10 days. Our results show that nickel induces atrophy of the seminiferous tubules with a reduction of the sperm count and a reduction of serum testosterone levels. A reduction of the number of ovarian follicles was observed in females. Intermittent fasting induced the same types of disturbances with more marked reductions of the number of mobile spermatozoa and serum festosterone levels than those observed after exposure to nickel. The combination of the two factors, fasting and nickel, did not amplify these effects. Analysis of intergroup crosses showed that the pregnancy rate and especially the mean number of implantations were decreased in rats exposed to nickel and/or submitted to intermittent fasting. The lowest pregnancy rate (55%) was observed in (NNi) females crossed with (NO) control males. The smallest number of implantations was observed in (NO) control females crossed with (NNi) males. Nickel did not induce any additional reduction of fertility in rats submitted to intermittent fasting. MDA assays showed that nickel induces lipid peroxidation in ovarian and uterine tissues. However, the relative increase of the MDA level was lower in FNi than NNi rats, i.e. when nickel was associated with intermittent fasting. Our results suggest that nickel and intermittent fasting decrease fertility in rats via two different mechanisms whose effects are not additive. When associated with intermittent fasting, nickel becomes non-toxic, as confirmed by montoring of MDA levels. The low-calorie effect of intermittent fasting could be responsible for inhibition of the cytotoxic effects of metallic nickel classified as an oxidative stress.     

Effects of nickel poisoning on expression pattern of the 72/73 and 94 kDa stress proteins in rat organs and in the COS-7, HepG2, and A549 cell lines

The present study deals with the effects of Ni on the expression level of three stress proteins, namely, the cytosolic HSP72 and HSP73, and the reticulum-associated GRP94. Experiments were carried out on "Wistar' female rats daily injected with 4 mg NiCl2 per kg body weight for 1, 3, 5, and 10 days. Another set of experiments were carried out using cell lines, derived from the monkey kidney (COS-7), and from human tumors of the lung (A549) and liver (HepG2). Cells were cultured for 4 days in the permanent presence of 100, 200, or 400 microM NiCl2. In control rats, stress proteins pattern was found to be tissue specific: two protein bands of 96 and 94 kDa were immunodetected with the anti-GRP94 antibody in kidney and liver extracts, whereas only the 96 kDa band was present in ovary extracts. HSP73 was present in kidney, liver, and ovary whereas HSP72 was only found in kidney. In kidney of nickel-treated animals, HSP73 and the 96 kDa proteins were overexpressed whereas HSP72 was strongly down regulated. No such effect was observed in liver or ovary. Similarly, in nickel-treated cell lines, HSP72 was downregulated and GRP94 (96 kDa protein) was overexpressed. HSP73 expression appeared moderately increased in A549 cells but decreased in COS-7 cells. Because long-term caloric restriction was reported to reduce free radical generation in cells, the effect of 1 month food restriction (50%) was tested in rats as a possible way to lower oxidative damages induced by Ni. No significant effect on HSP expression was observed.