GnRH Agonist (Buserelin) Induced In Vitro Apoptosis in Bovine Endometrium.
Notice bibliographique
Résumé
Apoptosis is a vital biological process that maintains tissue homeostasis by selective elimination of cells. The GnRH regulates gonadal physiology by acting as local modulator of apoptosis in ovaries, testes and prostate. Recently, we elucidated the presence of GnRH-R in the follicular and luteal phase bovine endometrium. The local modulatory role of GnRH, GnRH-R system in uterine biology is still not clear. Apoptosis acquires more significance in renewable structures like endometrium, undergoing cycles of cellular proliferation and elimination during the reproductive process. Therefore, we for the first time studied GnRH agonist (buserelin), induced apoptosis at the molecular (Bax, Bcl-2, Caspase, Fas mRNA), and the cellular level in the follicular and luteal phase bovine endometrium. 100 mg of endometrial explants (1-2 mm2) obtained from slaughter house reproductive tracts were cultured at 37°C in air with 5% CO2 in Dulbecco's modified eagle's medium containing 0.1% BSA, 100 IU/mL penicillin G and 100 μg/mL streptomycin for 20 h, then replaced media and treated the tissues with buserelin (0, 200, 500, 1000 ng/mL respectively), GnRH antagonist- antide (500 ng/mL) and antide + buserelin (500+200 ng/mL) for 6 h and stored the tissues at -80°C for RNA extraction. Two μg of total RNA was reverse transcribed using gene-specific primers and the PCR products were electrophoresed and visualized under UV illumination. The relative mRNA levels were measured using computerized software (Scion imaging beta) and normalized to the housekeeping gene G3PDH. To further study apoptosis at the cellular level, endometrial epithelial cells from the follicular and luteal phase endometrium were isolated, tested for cell viability using trypan blue exclusion method and seeded at a density of 100,000 cells/chamber in a collagen coated eight chamber glass slide at 37°C, 5% CO2 in DMEM/Ham F-12 medium (1:1), containing 10% heat-inactivated calf serum and 20 mg/ml gentamycin for 48 h. The cells were characterized by immunofluoroscence and using anticyokeratin as primary and FITC-labeled secondary antibody. After 48 h medium was replaced containing 1% calf serum and gentamycin and treated the cells with buserelin (200 ng/ml), antide (500 ng/ml) and a combination of antide + buserelin (500 + 200 ng/ml) for 6 h and allowed the cells to further grow for 24 h. The monolayers were then stained using acridine orange ethidium bromide and visualized under fluorescent microscope. Live and apoptotic cells were counted in three randomly selected fields in each chamber. The data were statistically analyzed using ANOVA and Student t and Dunett test (Caspase mRNA). This study revealed that buserelin (200 ng/ml) upregulates (P≤0.05) Bax and Caspase in follicular phase endometrium without affecting Bcl-2 and Fas mRNA levels. The cells in the present culture system were confirmed to be endometrial epithelial cells by observing clear labeling against cytokeratin. Buserelin (200 ng/mL) induced apoptosis (P≤0.05) in follicular endometrial epithelial cells. Luteal phase endometrium did not show apoptotic response to similar buserelin treatments at molecular or cellular level. This study for the first time demonstrate that GnRH agonist, buserelin induces receptor mediated apoptosis in follicular phase bovine endometrium at the molecular and cellular level and further indicates the involvement of mitochondrial cell death pathway in this phenomenon. (poster)
Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.
Comment cette classification a été obtenuedéplier
Prédiction machine sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Le volet Gemma est une étiquette directe du modèle pour chaque travail de la base, lue sur la notice réduite au titre. Le volet Codex est un classifieur appris des 10 348 étiquettes directes de Codex et calibré sur les taux pondérés de l'échantillon; les champs sans appui suffisant ne portent aucun appel Codex. Le mode candidate est l'union des deux volets; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont pas des étiquettes humaines.
Scores du classifieur distillé par catégorie (deux têtes)
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,000 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,002 | 0,001 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule source (Gemma direct ou Codex distillé), pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».