Hazardous Effects of Curcumin on Mouse Embryonic Development through a Mitochondria-Dependent Apoptotic Signaling Pathway

In this study, we examined the cytotoxic effects of curcumin, the yellow pigment of Curcuma longa, on the blastocyst stage of mouse embryos, subsequent embryonic attachment, and outgrowth in vitro and in vivo implantation by embryo transfer. Mouse blastocysts were incubated in medium with or without curcumin (6, 12 or 24 μM) for 24 h. Cell proliferation and growth were investigated using dual differential staining, apoptosis was analyzed with terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), and implantation and post-implantation development of embryos were measured by in vitro development analysis and in vivo embryo transfer, respectively. Blastocysts treated with 24 μM curcumin displayed significantly increased apoptosis and decreased total cell number. Interestingly, we observed no marked differences in the implantation success rates between curcumin-pretreated and control blastocysts during in vitro embryonic development through implantation with a fibronectin-coated culture dish. However, in vitro treatment with 24 μM curcumin was associated with decreased implantation rate and increased resorption of postimplantation embryos in mouse uterus, as well as decreased fetal weight in the embryo transfer assay. Our results collectively indicate that in vitro exposure to curcumin triggers apoptosis and retards early postimplantation development after transfer to host mice. In addition, curcumin induces apoptotic injury effects on mouse blastocysts through ROS generation, and further promotes mitochondria-dependent apoptotic signaling processes to impair sequent embryonic development

Effects of tnf-alpha on mouse embryo development

Le TNF-α (Tumor necrosis factor-α) est une cytokine exprimée dans de nombreux organes et de nombreux types de cellules, y compris l’utérus. On connaît peut de choses concernant ses effets sur le développement embryonnaire. Dans le présent travail, nous avons traité des blastocytes de souris avec du TNF-α recombinant durant 24h, et examines plusieurs paramètres de développement. Par rapport à des embryons contrôles, les blastocytes traités au TNF-α contenaient moins de cellules et le déficit touchait principalement la masse cellulaire interne (ICM). Le TNF-α augmentait l’incidence de cellules dans les noyaux montraient un processus de dégradation de la chromatine (karyolyse), sans influence sur l’incidence de fragmentation nucléaire (karyorrhexis). Une diminution de la prolifération et une augmentation de la karyolyse étaient également observées dans des cellules souches embryonnaires (ES) de souris exposées au TNF-α. Contrairement aux blastocytes, qui expriment uniquement la forme Rp60 du récepteur de TNF-α, les cellules ES expriment les deux isoforms Rp60 et Rp80. la blocage simultané des deux isoformes par des antagonistes était nécessaire pour supprimer l’effet du TNF-α sur le prolifération des cellules ES, indiquant que les deux isoformes contribuent à l’effet inhibiteur du TNF-α. Dans les expériences d’implantation in vitro, des blastocytes traités au TNF-α formaient des excroissances de surface identique à celles d’embryons non traits. Cependant, un plus petit nombre d’embryons traités par TNF-α étaient capables de maintenir un ICM compact et le nombre de cellules par embryon implanté était plus petit que dans le groupe contrôle. Il n’y avait pas d’évidence d’apoptose persistante dans les embryons implantés. Après transfert dans des souris porteuses, des blastocytes prétraités au TNF-α s’implantaient avec la même fréquence que des embryons contrôles mais plus d’embryons étaient résorbés au cours du développement ultérieur. Des cellules ES, exposées au TNF-α sur-exprimaient le gène Rex-1, marqueur de l’état indifférencié, et formaient moins de corps embryoides (EB), après transfert dans un système de culture en suspension montrant que le TNF-α avait altéré le potentiel de différenciation de ces cellules souches. Dans l’ensemble, notre étude montre que l’impact du TNF-α sur le blastocyte est d’inhiber aussi bien la prolifération que la différenciation et que l’ICM est la principale cible de la cytokine au moment de l’implantationThèse de doctorat en sciences biomédicales (Biologie de la reproduction) -- UCL, 199

Characterization of Apoptosis Induced by Emodin and Related Regulatory Mechanisms in Human Neuroblastoma Cells

Emodin (1,3,8-trihydroxy-6-methylanthraquinone), a major constituent of rhubarb, has a wide range of therapeutic applications. Recent studies have shown that emodin can induce or prevent cell apoptosis, although the precise molecular mechanisms underlying these effects are unknown. Experiments from the current study revealed that emodin (10–20 μM) induces apoptotic processes in the human neuroblastoma cell line, IMR-32, but exerts no injury effects at treatment doses below 10 μM. Treatment with emodin at concentrations of 10–20 μM led to a direct increase in the reactive oxygen species (ROS) content in IMR-32 cells, along with significant elevation of cytoplasmic free calcium and nitric oxide (NO) levels, loss of mitochondrial membrane potential (MMP), activation of caspases-9 and -3, and cell death. Pretreatment with nitric oxide (NO) scavengers suppressed the apoptotic biochemical changes induced by 20 μM emodin, and attenuated emodin-induced p53 and p21 expression involved in apoptotic signaling. Our results collectively indicate that emodin at concentrations of 10–20 μM triggers apoptosis of IMR-32 cells via a mechanism involving both ROS and NO. Based on the collective results, we propose a model for an emodin-triggered apoptotic signaling cascade that sequentially involves ROS, Ca2+, NO, p53, caspase-9 and caspase-3

Ochratoxin a inhibits mouse embryonic development by activating a mitochondrion-dependent apoptotic signaling pathway

Abstract: Ochratoxin A (OTA), a mycotoxin found in many foods worldwide, causes nephrotoxicity, hepatotoxicity, and immunotoxicity, both in vitro and in vivo. In the present study, we explored the cytotoxic effects exerted by OTA on the blastocyst stage of mouse embryos, on subsequent embryonic attachment, on outgrowth in vitro, and following in vivo implantation via embryo transfer. Mouse blastocysts were incubated with or without OTA (1, 5, or 10 μM) for 24 h. Cell proliferation and growth were investigated using dual differential staining; apoptosis was measured using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay; and embryo implantation and post-implantation development were assessed by examination of in vitro growth and the outcome of in vivo embryo transfer, respectively. Blastocysts treated with 10 μM OTA displayed a significantly increased level of apoptosis and a reduction in total cell number. Interestingly, we observed no marked difference in implantation success rate between OTA-pretreated and control blastocysts either during in vitro embryonic development (following implantation in a fibronectin-coated culture dish) or after in vivo embryo transfer. However, in vitro treatment with 10 μM OTA wasInt. J. Mol. Sci. 2013, 14 93

Two-Stage Fermented Feather Meal-Soybean Meal Product Improves the Performance and Immunity of Lactating Sows and Piglets

This study aimed to investigate the effects of a two-stage fermented feather meal-soybean meal product (TSFP) on the performance, clinical blood biochemistry, and immunity of sows and piglets. TSFP was fermented by Saccharomyces cerevisiae Y10 for three days in the second stage, which showed similar results to the five-day fermentation of B. coagulans (p > 0.05). Fifty hybrid sows (Duroc × KHAPS black pig) were randomly assigned into dietary supplementation groups of 2% fish meal or different levels of TSFP at 0%, 1%, 2%, or 3%. The results showed that body weight gain and feed conversion ratio of 2% and 3% TSFP groups were better than the control group and fish meal group during the gestation period (d 80–114) (p p p p p p < 0.05). In conclusion, TSFP supplementation exhibits the advantages of performance and immunity of lactating sows and piglets. Furthermore, adding 3% TSFP in the feed showed the best performance

The Effects of Two-Stage Fermented Feather Meal-Soybean Meal Product on Growth Performance, Blood Biochemistry, and Immunity of Nursery Pigs

The keratinase-producing microbes can readily change the feather waste into more bioavailable peptides and amino acids. This study aimed to improve two-stage fermented feather meal-soybean meal product (TSFP) quality with five selected Bacillus strains and Saccharomyces cerevisiae Y10, as well as investigate the effects of TSFP on growth performance, blood biochemistry, and immunity of nursery pigs. In trial 1, 100 hybrid piglets (Duroc x KHAPS) were randomly assigned to dietary supplementation of 5% fish meal or 5% TSFP prepared with 0, 32, 40, or 48 h in the first-stage fermentation. The results showed that the body weight (BW), average daily gain (ADG), and feed conversion ratio (FCR) of fermented groups were significantly better than the unfermented group (p &lt; 0.05) at weeks 0 to 3 and 0 to 5. The ADG of 32-hr and 48-hr TSFP groups were better than the unfermented group (p &lt; 0.05) at weeks 3 to 5. In trial 2, 80 hybrid piglets (Duroc &times; KHAPS) were randomly assigned into 5% fish meal or different supplementation levels of TSFP (32-hr first-stage fermented time) at 0, 2.5, or 5%. The 5% TSFP group had better BW, ADG, FCR, and PEF than the 0% group (p &lt; 0.05) at weeks 0 to 5. Furthermore, the ex vivo mitogen-induced lymphoblastogenesis, the interferon-&gamma; production, the oxidative burst activity, and the IgG production of the 5% TSFP group were higher than the fish meal group (p &lt; 0.05). In conclusion, the first-stage fermentation time can be shortened from 48 h to 32 h using selected Bacillus strains in TSFP production when supplemented at 5% of the diet for nursery pigs shows the best growth performance and immunity

The Effects of Two-Stage Fermented Feather Meal-Soybean Meal Product on Growth Performance, Blood Biochemistry, and Immunity of Nursery Pigs

The keratinase-producing microbes can readily change the feather waste into more bioavailable peptides and amino acids. This study aimed to improve two-stage fermented feather meal-soybean meal product (TSFP) quality with five selected Bacillus strains and Saccharomyces cerevisiae Y10, as well as investigate the effects of TSFP on growth performance, blood biochemistry, and immunity of nursery pigs. In trial 1, 100 hybrid piglets (Duroc x KHAPS) were randomly assigned to dietary supplementation of 5% fish meal or 5% TSFP prepared with 0, 32, 40, or 48 h in the first-stage fermentation. The results showed that the body weight (BW), average daily gain (ADG), and feed conversion ratio (FCR) of fermented groups were significantly better than the unfermented group (p p p p < 0.05). In conclusion, the first-stage fermentation time can be shortened from 48 h to 32 h using selected Bacillus strains in TSFP production when supplemented at 5% of the diet for nursery pigs shows the best growth performance and immunity