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1.
Hum Reprod ; 39(2): 403-412, 2024 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-38110714

RESUMO

STUDY QUESTION: How do plasma progesterone (P) and dydrogesterone (D) concentrations together with endometrial histology, transcriptomic signatures, and immune cell composition differ when oral dydrogesterone (O-DYD) or micronized vaginal progesterone (MVP) is used for luteal phase support (LPS)? SUMMARY ANSWER: Although after O-DYD intake, even at steady-state, plasma D and 20αdihydrodydrogesterone (DHD) concentrations spiked in comparison to P concentrations, a similar endometrial signature was observed by histological and transcriptomic analysis of the endometrium. WHAT IS KNOWN ALREADY: O-DYD for LPS has been proven to be noninferior compared to MVP in two phase III randomized controlled trials. Additionally, a combined individual participant data and aggregate data meta-analysis indicated that a higher pregnancy rate and live birth rate may be obtained in women receiving O-DYD versus MVP for LPS in fresh IVF/ICSI cycles. Little data are available on the pharmacokinetic (PK) profiles of O-DYD versus MVP and their potential molecular differences at the level of the reproductive organs, particularly at the endometrial level. STUDY DESIGN, SIZE, DURATION: Thirty oocyte donors were planned to undergo two ovarian stimulation (OS) cycles with dual triggering (1.000 IU hCG + 0.2 mg triptorelin), each followed by 1 week of LPS: O-DYD or MVP, in a randomized, cross-over, double-blind, double-dummy fashion. On both the first and eighth days of LPS, serial blood samples upon first dosing were harvested for plasma D, DHD, and P concentration analyses. On Day 8 of LPS, an endometrial biopsy was collected for histologic examination, transcriptomics, and immune cell analysis. PARTICIPANTS/MATERIALS, SETTING, METHODS: All oocyte donors were <35 years old, had regular menstrual cycles, no intrauterine contraceptive device, anti-Müllerian hormone within normal range and a BMI ≤29 kg/m2. OS was performed on a GnRH antagonist protocol followed by dual triggering (1.000 IU hCG + 0.2 mg triptorelin) as soon as ≥3 follicles of 20 mm were present. Following oocyte retrieval, subjects initiated LPS consisting of MVP 200 mg or O-DYD 10 mg, both three times daily. D, DHD, and P plasma levels were measured using liquid chromatography-tandem mass spectrometry. Histological assessment was carried out using the Noyes criteria. Endometrial RNA-sequencing was performed for individual biopsies and differential gene expression was analyzed. Endometrial single-cell suspensions were created followed by flow cytometry for immune cell typing. MAIN RESULTS AND THE ROLE OF CHANCE: A total of 21 women completed the entire study protocol. Subjects and stimulation characteristics were found to be similar between groups. Following the first dose of O-DYD, the average observed maximal plasma concentrations (Cmax) for D and DHD were 2.9 and 77 ng/ml, respectively. The Cmax for D and DHD was reached after 1.5 and 1.6 h (=Tmax), respectively. On the eighth day of LPS, the first administration of that day gave rise to a Cmax of 3.6 and 88 ng/ml for D and DHD, respectively. For both, the observed Tmax was 1.5 h. Following the first dose of MVP, the Cmax for P was 16 ng/ml with a Tmax of 4.2 h. On the eighth day of LPS, the first administration of that day showed a Cmax for P of 21 ng/ml with a Tmax of 7.3 h. All 42 biopsies showed endometrium in the secretory phase. The mean cycle day was 23.9 (±1.2) in the O-DYD group versus 24.0 (±1.3) in the MVP group. RNA-sequencing did not reveal significantly differentially expressed genes between samples of both study groups. The average Euclidean distance between samples following O-DYD was significantly lower than following MVP (respectively 12.1 versus 18.8, Mann-Whitney P = 6.98e-14). Immune cell profiling showed a decrease of CD3 T-cell, γδ T-cell, and B-cell frequencies after MVP treatment compared to O-DYD, while the frequency of natural killer (NK) cells was significantly increased. LIMITATIONS, REASONS FOR CAUTION: The main reason for caution is the small sample size, given the basic research nature of the project. The plasma concentrations are best estimates as this was not a formal PK study. Whole tissue bulk RNA-sequencing has been performed not correcting for bias caused by different tissue compositions across biopsies. WIDER IMPLICATIONS OF THE FINDINGS: This is the first study comparing O-DYD/MVP, head-to-head, in a randomized design on a molecular level in IVF/ICSI. Plasma serum concentrations suggest that administration frequency is important, in addition to dose, specifically for O-DYD showing a rapid clearance. The molecular endometrial data are overall comparable and thus support the previously reported noninferior reproductive outcomes for O-DYD as compared to MVP. Further research is needed to explore the smaller intersample distance following O-DYD and the subtle changes detected in endometrial immune cells. STUDY FUNDING/COMPETING INTEREST(S): Not related to this work, C.Bl. has received honoraria for lectures, presentations, manuscript writing, educational events, or scientific advice from Abbott, Ferring, Organon, Cooper Surgical, Gedeon-Richter, IBSA, and Merck. H.T. has received honoraria for lectures, presentations, manuscript writing, educational events, or scientific advice from Abbott, Ferring, Cooper Surgical, Gedeon-Richter, Cook, and Goodlife. S.M. has received honoraria for lectures, presentations, educational events, or scientific advice from Abbott, Cooper Surgical, Gedeon-Richter, IBSA, and Merck and Oxolife. G.G. has received honoraria for lectures, presentations, educational events, or scientific advice from Merck, MSD, Organon, Ferring, Theramex, Gedeon-Richter, Abbott, Biosilu, ReprodWissen, Obseva, PregLem, Guerbet, Cooper, Igyxos, and OxoLife. S.V.-S. is listed as inventor on two patents (WO2019115755A1 and WO2022073973A1), which are not related to this work. TRIAL REGISTRATION NUMBER: EUDRACT 2018-000105-23.


Assuntos
Didrogesterona , Progesterona , Gravidez , Humanos , Feminino , Adulto , Estudos Cross-Over , Pamoato de Triptorrelina , Fase Luteal , Lipopolissacarídeos , Injeções de Esperma Intracitoplásmicas/métodos , Taxa de Gravidez , Indução da Ovulação/métodos , Endométrio , RNA , Fertilização in vitro/métodos , Ensaios Clínicos Controlados Aleatórios como Assunto
2.
Homeopathy ; 113(1): 4-15, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37517405

RESUMO

INTRODUCTION: Aspirin is one of the most commonly used drugs worldwide. It is known to present antipyretic, anti-inflammatory and anti-thrombotic actions, making it extremely useful in a wide range of clinical contexts. Interestingly, homeopathically prepared Aspirin 15cH has been found to have a pro-thrombotic effect in rats, raising the hypothesis that Aspirin 15cH could also modulate the activity of inflammatory cells in different pathological processes. OBJECTIVE: Our objective was to assess what effect Aspirin 15cH has on RAW 264.7 macrophages in vitro. METHODS: The effects of Aspirin 15cH on biochemical and morphological activities of lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages were evaluated. These effects were compared with unchallenged macrophages (negative control), untreated LPS-stimulated macrophages, macrophages treated with succussed water (vehicle control), or aspirin 200 µg/mL (pharmacological inhibitor of LPS activity). Cell morphology (adhered cell area and cytoskeleton arrangements), cell viability, toll-like receptor-4 (TLR-4) expression, and the production of nitric oxide, cytokines and intracellular reactive oxygen species were assessed. RESULTS: Aspirin 15cH reduced the number of cells expressing TLR-4 on the surface (p = 0.03) and induced a "columnar" morphology of macrophage pseudopods, indicating changes in cytoskeleton arrangement. When cells were treated with both Aspirin 15cH and LPS, cell morphology became heterogeneous, suggesting that sub-populations of cells had differing sensitivities to LPS or Aspirin 15cH. Exposure of the cells to LPS alone, succussed water or aspirin 200 µg/mL produced effects consistent with the literature. CONCLUSION: Aspirin 15cH, aspirin 200 µg/mL, LPS and succussed water appear to act as independent stimuli able to induce different patterns of macrophage response. Aspirin 15cH induced changes suggestive of M2 polarization of the macrophages (i.e., toward a wound healing or tissue repair, rather than inflammatory, phenotype). These preliminary findings need to be confirmed in further specific studies.


Assuntos
Homeopatia , Lipopolissacarídeos , Ratos , Animais , Lipopolissacarídeos/farmacologia , Lipopolissacarídeos/metabolismo , Aspirina/farmacologia , Receptor 4 Toll-Like/metabolismo , Macrófagos , Citocinas , Água
3.
Zhongguo Zhong Xi Yi Jie He Za Zhi ; 30(6): 625-9, 2010 Jun.
Artigo em Zh | MEDLINE | ID: mdl-20815280

RESUMO

OBJECTIVE: To investigate the protective effects of musk extract (ME) and its possible mechanism on rat's cerebral cortical neurons with inflammatory injury induced by lipopolysaccharide (LPS). METHODS: Neurons and astrocytes from newborn rat cerebral cortex were cultured in vitro respectively, and the astrocyte conditioned medium (ACM), obtained by treating astrocytes with 10 mg/L LPS and different concentrations of ME for 24 h, was added in the culture fluid of neurons. The survival rate and apoptotic rate of neurons were measured by MTT method and AO/EB stain; and the changes of inflammatory factors in the ACM were determined by ELISA. RESULTS: The survival rate (%) of neurons treated by ACM with ME in concentrations of 18 mg/L, 36 mg/L, 72 mg/L and 144 mg/L was 52.55 +/- 3.52, 55.77 +/- 2.36, 64.89 +/- 3.45 and 73.67 +/- 1.80, respectively, significantly higher than that in the model neurons (43.62 +/- 4. 51, P < 0.05), while the apoptotic rate (%) in them, 68.11 +/- 2.16, 44.27 +/- 3.68, 32.56 +/- 2.14 and 21.89 +/- 2.46, respectively, was significantly lower than that in model neurons (71.33 +/- 3.25, P < 0.05 or P < 0.01). Level of IL-6 was decreasing along with the raising of ME concentration in the ACM, showing a concentration-dependent state. CONCLUSION: ME shows apparent protective effect on neurons against inflammatory injury, especially in a high concentration (144 mg/L), which may be associated with the reduction of IL-6 secreted by astrocytes.


Assuntos
Córtex Cerebral/citologia , Ácidos Graxos Monoinsaturados/química , Inflamação/prevenção & controle , Neurônios/citologia , Substâncias Protetoras/farmacologia , Animais , Animais Recém-Nascidos , Astrócitos/citologia , Astrócitos/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Inflamação/induzido quimicamente , Interleucina-6/metabolismo , Lipopolissacarídeos , Masculino , Materia Medica/farmacologia , Ratos , Ratos Wistar
4.
Eur Cytokine Netw ; 18(4): 210-4, 2007 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17993452

RESUMO

There are a few studies with conflicting results on the effects of opioids on the functioning of immune system. This study was performed to investigate the in vitro production of interferon-gamma and interleukin-10 after antigenic stimulation of cells using whole blood from opioid addicts. Blood samples were taken from 20 chronically opioid-addicted persons, who voluntarily enrolled for detoxification (10 opium and 10 heroin addicts). Blood samples were also taken from 10 healthy individuals with no history of drug abuse as the control. Cell culture was performed in a whole blood culture assay. Diluted blood samples were stimulated with phytohemagglutinin or with lipopolysaccharide and the supernatants were collected to measure cytokine production. The results demonstrated a significant decrease in interferon-gamma production and an increase in interleukin-10 secretion in heroin addicts, relative to the control group (35.9+/-26.3 versus 110.2+/-60.3 pg/mL, p<0.01 and 71.8+/-28.4 versus 17.1+/-13.5 pg/mL, p<0.01, respectively), however the changes in these values in opium addicts were not significant compared to healthy individuals. The results could suggest that opioid addiction leads to a shift in the Th1/Th2 cytokine balance of peripheral CD4+ cells towards the Th2 response, and opioid addicts demonstrate reduced mitogenic responsiveness of lymphocytes relative to healthy individuals.


Assuntos
Analgésicos Opioides/metabolismo , Citocinas/metabolismo , Células Th1/metabolismo , Células Th2/metabolismo , Adulto , Antígenos/metabolismo , Linfócitos T CD4-Positivos/metabolismo , Hemaglutininas/química , Heroína/metabolismo , Humanos , Interleucina-10/metabolismo , Lipopolissacarídeos/metabolismo , Mitógenos/metabolismo , Ópio/metabolismo , Transtornos Relacionados ao Uso de Substâncias
5.
Acta Physiol Hung ; 87(2): 161-6, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-11205964

RESUMO

Glucocorticoids are important modulators of immune reactions. They are capable of antagonising several effects of the bacterial endotoxin by inhibiting endotoxin-induced leukocyte activation, and the production of cytokines and inflammatory mediators. We earlier demonstrated that the antiglucocorticoid RU 38486 enhances the cytokine production induced by endotoxin and aggravates the course of experimental endotoxic and septic shock. In the present study we investigated the effect of the glucocorticoid Oradexon on the endotoxin-induced peritoneal cell response. For measurement of the peritoneal cell response, male CFLP mice (20-25 g) were injected i.p. with 10 microg/10 g body weight endotoxin (E. coli 026:B6 LPS, Difco Lab, Detroit, lot 110273JB). Dexamethasone (Oradexon, N.V, Organon Oss, The Netherlands) was administered i.p., i.v. or s.c. in a dose of 0.1 mg/10 g body weight, alone or concomitantly with endotoxin. We found that bacterial endotoxin increased the total cell count due to neutrophilia at 24 hours and, due to increases in the number of macrophages and lymphocytes 48 and 72 hours after treatment, respectively. The i.p., i.v., and s.c. injection of Oradexon, increased the total cell count and the macrophage count at 24, 48 and 72 hours. The i.p., s.c. and i.v. injection of Oradexon, concomitantly with endotoxin, reduced the total cell count at 48 and 72 hours, due to decreases in the macrophage count. The i.p., i.v. or s.c. administration of Oradexon concomitantly with LPS decreased the lymphocyte count and the neutrophil count at 24 and 72 hours. These results prove that glucocorticoids are capable of modifying the immune cell reactions induced by endotoxin.


Assuntos
Anti-Inflamatórios/farmacologia , Dexametasona/farmacologia , Endotoxinas/antagonistas & inibidores , Imunidade Celular/efeitos dos fármacos , Mediadores da Inflamação/metabolismo , Ativação de Macrófagos/efeitos dos fármacos , Animais , Anti-Inflamatórios/administração & dosagem , Dexametasona/administração & dosagem , Endotoxinas/toxicidade , Granulócitos/efeitos dos fármacos , Granulócitos/imunologia , Injeções Intraperitoneais , Injeções Intravenosas , Injeções Subcutâneas , Lipopolissacarídeos/antagonistas & inibidores , Lipopolissacarídeos/toxicidade , Linfócitos/efeitos dos fármacos , Linfócitos/imunologia , Masculino , Camundongos , Infiltração de Neutrófilos/efeitos dos fármacos , Cavidade Peritoneal/citologia
6.
Homeopathy ; 93(4): 193-8, 2004 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-15532698

RESUMO

Atropa belladonna and Echinacea angustifolia have been used in homeopathy as modulators of inflammatory processes, in simple potency or 'accord of potencies', as recommended by homotoxicology. We evaluated their effects on leukocyte migration and macrophage activity induced by experimental peritonitis in vivo. Mice were injected (i.p.) with LPS (1.0mg/kg) and treated (0.3ml/10g/day, s.c.) with different commercial forms of these medicines. Echinacea angustifolia D4--a simple potency preparation--and Belladonna Homaccord, Belladonna Injeel, Belladonna Injeel Forte, Echinacea Injeel and Echinacea Injeel Forte--all in 'accord of potencies'--were tested. The association of A. belladonna and E. angustifolia in 'accord of potencies' produced an increase of polymorphonuclear cell migration (Kruskal-Wallis, P = 0.03) and a decrease of mononuclear cell percentages (Kruskal-Wallis, P < or = 0.04), when compared with control, mainly in preparations containing low potencies. The proportion of degenerate leukocytes was lower in the treated groups, compared to a control group (P < or = 0.05). The treated groups showed increased phagocytosis (P < or = 0.05), mainly in preparations containing high potencies. Our results suggest that A. belladonna and E. angustifolia, when prepared in 'accord of potencies', modulate peritoneal inflammatory reaction and have a cytoprotective action on leukocytes.


Assuntos
Anti-Inflamatórios/farmacologia , Atropa belladonna , Echinacea , Homeopatia/métodos , Leucócitos/efeitos dos fármacos , Peritonite/tratamento farmacológico , Animais , Inibição de Migração Celular , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Leucócitos/imunologia , Lipopolissacarídeos , Macrófagos Peritoneais/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Peritonite/induzido quimicamente , Extratos Vegetais/farmacologia , Distribuição Aleatória
7.
Biomed Sci Instrum ; 39: 371-6, 2003.
Artigo em Inglês | MEDLINE | ID: mdl-12724922

RESUMO

For centuries, people in the Middle East and Southeast Asia have used Nigella sativa, also known as black seed (BS), for its homeopathic effects. The objective of this study was to investigate the role BS might have on the metabolic biomarkers of the Hep-2 cell line. The experimental design entailed six groups of five wells each (50,000 cells). Groups II through VI were treated with BS, lipopolysaccharide (LPS), cortisol, LPS + cortisol, and BS + LPS + cortisol, respectively. Group I was the untreated control group. At the end of 24, 48, and 72 hours, the total cell count, protein and MDA levels were measured by following standard lab protocols. Data collected from this study revealed that Hep-2 cells exposed to LPS and cortisol (group V) resulted in a decrease in cell proliferation compared to the control. BS treatment induced a higher proliferation rate than group V. Similar trends were observed in the metabolic behavior of Hep-2 cells as evidenced by the total protein and MDA levels. The exposure of BS showed a shift in the metabolic pathways. In conclusion, this study showed that exposure to LPS resulted in an alteration in the metabolic function and this phenomenon was further escalated under stressful conditions (increased cortisol exposure). In addition, the use of BS reversed the traumatic condition.


Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias Laríngeas/tratamento farmacológico , Neoplasias Laríngeas/metabolismo , Nigella sativa , Extratos Vegetais/uso terapêutico , Divisão Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Humanos , Hidrocortisona/administração & dosagem , Neoplasias Laríngeas/patologia , Neoplasias Laríngeas/fisiopatologia , Lipopolissacarídeos/administração & dosagem , Malondialdeído/metabolismo , Fitoterapia/métodos , Proteínas/metabolismo , Valores de Referência , Sementes , Sensibilidade e Especificidade , Resultado do Tratamento , Células Tumorais Cultivadas
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