Effects of the Methanol Extract of Basella alba L (Basellaceae) on Steroid Production in Leydig Cells

In this study, Leydig cells were purified from 70 day-old Sprague Dawley male rats and incubated with 10 and 100 μg/mL of methanol extract of Basella alba (MEBa) for 4 hours followed by the evaluation of cell viability, steroid (testosterone and estradiol) production, and the level of aromatase mRNA. Results showed that MEBa did not affect Leydig cell viability. At the concentration of 10 μg/mL, MEBa significantly stimulated testosterone and estradiol production (p < 0.01 and p < 0.03, respectively), and enhanced aromatase mRNA level (p < 0.04). These observations suggest that MEBa directly stimulated testosterone, estradiol and aromatase mRNA levels in isolated Leydig cells

First synthesis of 7 beta- and 7 beta-amino-DHEA, dehydroepiandrosterone (DHEA) analogues and preliminary evaluation of their cytotoxicity on Leydig cells and TM4 sertoli cells

International audienc

Effects of Roundup® and Glyphosate on Three Food Microorganisms: Geotrichum candidum, Lactococcus lactis subsp. cremoris and Lactobacillus delbrueckii subsp. bulgaricus

International audienceUse of many pesticide products poses the problem of their effects on environment and health. Amongst them, the effects of glyphosate with its adjuvants and its by-products are regularly discussed. The aim of the present study was to shed light on the real impact on biodiversity and ecosystems of Roundup(®), a major herbicide used worldwide, and the glyphosate it contains, by the study of their effects on growth and viability of microbial models, namely, on three food microorganisms (Geotrichum candidum, Lactococcus lactis subsp. cremoris and Lactobacillus delbrueckii subsp. bulgaricus) widely used as starters in traditional and industrial dairy technologies. The presented results evidence that Roundup(®) has an inhibitory effect on microbial growth and a microbicide effect at lower concentrations than those recommended in agriculture. Interestingly, glyphosate at these levels has no significant effect on the three studied microorganisms. Our work is consistent with previous studies which demonstrated that the toxic effect of glyphosate was amplified by its formulation adjuvants on different human cells and other eukaryotic models. Moreover, these results should be considered in the understanding of the loss of microbiodiversity and microbial concentration observed in raw milk for many years

Stallion spermatozoa: putative targets of estrogens. Studies of estrogens receptors.

International audienceIntroduction. Among the mammals, stallion appears as the male producing the largest amount of testicular estrogens (Raeside, Can J Biochem., 47:811-5, 1969). This synthesis seems to be submitting to a seasonal regulation as Lemazurier et al. (Gen Comp Endocrinol. 125:272-82, 2002) demonstrated the presence of higher amounts of estrogens in semen in April-May and June compared to non-breeding season. The main sites of estrogen production are the Leydig cells, which show a strong immunoreactivity for aromatase (Almadhidi et al., J. Histochem Cytochem. 43:571-577, 1995). To exert their effects estrogens use specific nuclear receptor named ESR1 and ESR2, which could exert both genomic and non-genomic effects. Estrogen receptors have been localized in testis (Pearl et al., Anim Reprod Sci. 125 :103-11. 2011) as in epididymis (Parlevliet Theriogenology. 2006 ;66 :755-65). The presence of aromatase and estrogen receptors both in testis and in reproductive tract suggests that estrogen may play a role in spermatogenesis as well as in post-testicular maturation. To better define this role and the targets of estrogen we studied the putative presence of estrogen receptors on stallion spermatozoa.Materials and Methods. Semen was obtained from 10 stallions, aged from 10 to 23 years, housed at Jumenterie du Pin (IFCE). Seminal plasma was removed by centrifugation and spermatozoa were washed by two successive centrifugations in Tyrode. Analysis were performed by immunocytochemistry, Western-blot, immunofluorescence analysis and flow cytometry with MC-20 (ESR1), H-150 (ESR2) antibodies. GPER identification in equine specie was confirmed by RT-PCR and sequencing.Results. We showed the presence of both estrogen receptors ESR1, ESR2 on stallion spermatozoa. For ESR1, western-blot analysis shows a single 66kDa band corresponding to the wild-type isoform, immunofluorescence analysis showed a flagellar staining. For ESR2, western-blot analysis showed a single 61 kDa band, corresponding to the molecular weight described for the wild type form of this receptor and confocal analysis demonstrated also a flagellar localization. The rate of spermatozoa positive for the detection of these both receptors was analyzed by flow cytometry. We showed that 98 % of spermatozoa were positive for ESR1 and 94% positive for ESR2 in samples from 3 stallions obtained in April. The estrogen transmembrane receptor, GPER was also searched. Genome from Equus Caballus contains a GPER-like gene but there was no study about identification and expression of this gene in horse species. We proceeded to RNA extraction from a colt testis and spermatozoa from three stallions and with primers designed from the DNA sequence, we performed RT-PCR amplification. The products obtained were sequenced and allowed to describe the expression of GPER in equine species for the first time.Conclusion. We described in this study, the presence of estrogen receptors on stallion spermatozoa, allowing to describe it as a putative estrogen target. Moreover, we report the first description of GPER in equine species, opening new possibilities of studies of estradiol action in male as in female reproduction. Future studies will investigate putative use of these receptors to regulate spermatozoa functions like motility

Estrogènes et qualité de la semence chez le verrat

National audienc

Dig1 protects against cell death provoked by glyphosate-based herbicides in human liver cell lines

Abstract Background Worldwide used pesticides containing different adjuvants like Roundup formulations, which are glyphosate-based herbicides, can provoke some in vivo toxicity and in human cells. These pesticides are commonly found in the environment, surface waters and as food residues of Roundup tolerant genetically modified plants. In order to know their effects on cells from liver, a major detoxification organ, we have studied their mechanism of action and possible protection by precise medicinal plant extracts called Dig1. Methods The cytotoxicity pathways of four formulations of glyphosate-based herbicides were studied using human hepatic cell lines HepG2 and Hep3B, known models to study xenobiotic effects. We monitored mitochondrial succinate dehydrogenase activity and caspases 3/7 for cell mortality and protection by Dig1, as well as cytochromes P450 1A1, 1A2, 3A4 and 2C9 and glutathione-S-transferase to approach the mechanism of actions. Results All the four Roundup formulations provoke liver cell death, with adjuvants having stronger effects than glyphosate alone. Hep3B are 3-5 times more sensitive over 48 h. Caspases 3/7 are greatly activated in HepG2 by Roundup at non-cytotoxic levels, and some apoptosis induction by Roundup is possible together with necrosis. CYP3A4 is specifically enhanced by Roundup at doses 400 times less than used in agriculture (2%). CYP1A2 is increased to a lesser extent together with glutathione-S-transferase (GST) down-regulation. Dig 1, non cytotoxic and not inducing caspases by itself, is able to prevent Roundup-induced cell death in a time-dependant manner with an important efficiency of up to 89%, within 48 h. In addition, we evidenced that it prevents Caspases 3/7 activation and CYP3A4 enhancement, and not GST reduction, but in turn it slightly inhibited CYP2C9 when added before Roundup. Conclusion Roundup is able to provoke intracellular disruption in hepatic cell lines at different levels, but a mixture of medicinal plant extracts Dig1 can protect to some extent human cell lines against this pollutants. All this system constitutes a tool for studying liver intoxication and detoxification.</p