Developmental excitatory-to-inhibitory GABA-polarity switch is disrupted in 22q11.2 deletion syndrome: a potential target for clinical therapeutics.

Individuals with 22q11.2 microdeletion syndrome (22q11.2 DS) show cognitive and behavioral dysfunctions, developmental delays in childhood and risk of developing schizophrenia and autism. Despite extensive previous studies in adult animal models, a possible embryonic root of this syndrome has not been determined. Here, in neurons from a 22q11.2 DS mouse model (Lgdel +/-), we found embryonic-premature alterations in the neuronal chloride cotransporters indicated by dysregulated NKCC1 and KCC2 protein expression levels. We demonstrate with large-scale spiking activity recordings a concurrent deregulation of the spontaneous network activity and homeostatic network plasticity. Additionally, Lgdel +/- networks at early development show abnormal neuritogenesis and void of synchronized spontaneous activity. Furthermore, parallel experiments on Dgcr8 +/- mouse cultures reveal a significant, yet not exclusive contribution of the dgcr8 gene to our phenotypes of Lgdel +/- networks. Finally, we show that application of bumetanide, an inhibitor of NKCC1, significantly decreases the hyper-excitable action of GABAA receptor signaling and restores network homeostatic plasticity in Lgdel +/- networks. Overall, by exploiting an on-a-chip 22q11.2 DS model, our results suggest a delayed GABA-switch in Lgdel +/- neurons, which may contribute to a delayed embryonic development. Prospectively, acting on the GABA-polarity switch offers a potential target for 22q11.2 DS therapeutic intervention

Evaluation Of Pre-Freeze And Post-Thaw Sperm Quality Of Epididymal Canine Sperm Cooled In Situ Or Extended Overnight

- Introduction and objectives Sperm harvesting from the epididymal cauda represents the last chance to obtain progeny in case of an unforeseen decease or castration. However, not all small animal clinics perform sperm cryopreservation and epididymides need to be shipped to specialized facilities. Unfortunately, the lack of technical knowledge regarding its optimal storage and shipping conditions prevents the preservation of canine epididymal sperm. Hence, our study aimed to define the best cooling method for canine epididymal sperm prior cryopreservation at 4 ˚C: within the epididymis or extended. - Materials and methods Testicles were collected from 11 healthy dogs of different breeds immediately after castration. One epididymis was immediately stored at 4 ˚C for 24 h (Time 0). The contralateral epididymis was flushed with CaniPlus Chill®, extended to reach 150 × 106 sperm/ml and preserved at 4 ˚C. After 24 h of cooling, the stored epididymis was flushed and processed as the former. Samples were centrifuged, and the obtained pellet was resuspended at 150 × 106 sperm/ml in CaniPlus Freeze® medium, plus 20% egg yolk (v/v). Sperm were packed in 0.5 ml straws, cooled at 4 ˚C for 1 h, exposed to liquid nitrogen vapor for 20 minutes and plunged into liquid nitrogen. Sperm quality was assessed in fresh and frozen-thawed samples at all the time points: 0 and 24 h (fresh samples) and after thawing (37 ˚C for 1 min). Motility was evaluated using a CASA system; viability, mitochondrial membrane potential (MMP) and DNA integrity were assessed by flow cytometry using SYBR-14/PI, JC-1 and sperm chromatin structure respectively. Results are expressed as the mean ± SEM in % comparing epididymal cooling vs. extended sperm. ANOVA and student t-test were used to compare normally distributed data; Kruskal-Wallis test and Mood’s median test for non-normally distributed data; p < 0.05 was considered as significant. - Results Canine epididymal sperm, cooled for 24 h at 4 ˚C within the epididymis, showed significantly differences compared to the cooled-extended samples respectively for viability (69.9 ± 1.8 vs. 59.6 ± 3.4, mean % ± SEM; p < 0.05), higher MMP (64.9 ± 3.7 vs. 51.1 ± 2.7, mean % ± SEM; p < 0.05), and beat cross frequency (BCF) (7.5 ± 0.5 vs. 5.6 ± 0.6, mean % ± SEM; p < 0.05). After cryopreservation, significant differences were also found in MMP (50.9 ± 2.2 vs. 39.1 ± 4.9, mean % ± SEM; p < 0.05) and in viability (48.5 ± 2.7 vs. 39.5 ± 4.2, mean % ± SEM; p < 0.05) in samples cooled in the epididymis vs. extended 24 h prior cryopreservation, respectively. DNA fragmentation remained unaffected in all the treatments (p > 0.05). - Conclusions The study demonstrates that epididymal sperm better maintains its quality when cooled within the epididymis prior freezing. This finding opens a new horizon in the small animal clinics procedures to store and/or ship epididymal sperm in dogs prior freezing. Funding: RYC-2017-21545 (AEI/FEDER/UE, Spain); TA18008 (Junta de Extremadura, Spain)

Modulation of gonadotrophin induced steroidogenic enzymes in granulosa cells by d-chiroinositol

d-chiroinositol (DCI) is a inositolphosphoglycan (IPG) involved in several cellular functions that control the glucose metabolism. DCI functions as second messenger in the insulin signaling pathway and it is considered an insulin sensitizer since deficiency in tissue availability of DCI were shown to cause insulin resistance (IR). Polycystic ovary syndrome (PCOS) is a pathological condition that is often accompanied with insulin resistance. DCI can positively affects several aspect of PCOS etiology decreasing the total and free testosterone, lowering blood pressure, improving the glucose metabolism and increasing the ovulation frequency. The purpose of this study was to evaluate the effects of DCI and insulin combined with gonadotrophins namely follicle-stimulating hormone (FSH) and luteinizing hormone (LH) on key steroidogenic enzymes genes regulation, cytochrome P450 family 19 subfamily A member 1 (CYP19A1) and cytochrome P450 side-chain cleavage (P450scc) in primary cultures of human granulosa cells (hGCs). We also investigated whether DCI, being an insulin-sensitizer would be able to counteract the expected stimulator activity of insulin on human granulosa cells (hGCs)

Células madre y sus exosomas como terapia avanzada para tratar la hernia incisional: prueba de concepto en modelo murino

An incisional hernia constitutes a tissue protrusion through a traumatic or surgical scar in the abdominal wall. Frequently, the treatment of the incisional hernia, as well as other types of hernia, involves the implantation of a surgical mesh to reinforce the weakened tissue. However, an exacerbated inflammatory response is commonly developed after this implantation, having serious consequences for the patient. Considering the immunomodulatory potential of mesenchymal stem cells (MSCs) and their exosomes (exo-MSCs), in this study we proposed that the administration of these two therapeutic products, together with fibrin sealants that are frequently used to fix surgical meshes, could have a beneficial biological and therapeutic effect that could help to modulate the inflammatory response and improve the success of the surgical mesh implantation. The results obtained in this work showed, in a murine model of incisional hernia, that exo-MSCs reduce M1 inflammatory macrophages infiltration and that there is a predominance of Th2-related cytokines in the surrounding tissue of MSCs or exo-MSCs treated meshes, favoring the macrophage polarization towards a M2 anti-inflammatory phenotype. This study concludes that mesh fixation with fibrin sealants co-administered with MSCs or exo-MSCs would have a beneficiary effect on the treatment of incisional hernia in terms of reduction of the inflammatory response and modulation of the foreign body reaction.Una hernia incisional consiste en una protrusión de tejido a través de una cicatriz traumática o quirúrgica en la pared abdominal. El tratamiento de este y de otros tipos de hernias pasa frecuentemente por la implantación de una malla quirúrgica para reforzar el tejido debilitado. Sin embargo, a menudo se produce una respuesta inflamatoria exacerbada que desemboca en diferentes complicaciones, teniendo consecuencias graves para el paciente.Considerando el potencial inmunomodulador de las células madre mesenquimales (MSCs) y de sus exosomas (exo-MSCs), en este estudio planteamos que la administración de ambos productos terapéuticos, conjuntamente con los selladores de fibrina que se utilizan frecuentemente para la fijación de las mallas quirúrgicas, podría ejercer un efecto biológico y terapéutico que ayudara a controlar esa respuesta inflamatoria y mejorara, por tanto, el éxito del tratamiento con mallas quirúrgicas.Los resultados obtenidos en este estudio mostraron, en un modelo murino de hernia incisional, que los exo-MSCs reducen la infiltración de macrófagos inflamatorios M1 y que existe una predominancia de citoquinas relacionadas con la respuesta Th2, y con ello, con la polarización de macrófagos hacia un fenotipo M2 antiinflamatorio, en el tejido circundante a las mallas en las que se vehicularon MSCs o sus exosomas.Este estudio concluye que la fijación de mallas quirúrgicas con selladores de fibrina combinados con MSCs o exo-MSCs tendría un efecto beneficioso en el tratamiento de la hernia incisional, en términos de reducción de la respuesta inflamatoria y modulación de una reacción exacerbada frente a un cuerpo extraño

Intrapericardial Administration of Secretomes from Menstrual Blood-Derived Mesenchymal Stromal Cells: Effects on Immune-Related Genes in a Porcine Model of Myocardial Infarction.

Acute myocardial infarction (AMI) is a manifestation of ischemic heart disease where the immune system plays an important role in the re-establishment of homeostasis. We hypothesize that the anti-inflammatory activity of secretomes from menstrual blood-derived mesenchymal stromal cells (S-MenSCs) and IFNγ/TNFα-primed MenSCs (S-MenSCs*) may be considered a therapeutic option for the treatment of AMI. To assess this hypothesis, we have evaluated the effect of S-MenSCs and S-MenSCs* on cardiac function parameters and the involvement of immune-related genes using a porcine model of AMI. Twelve pigs were randomly divided into three biogroups: AMI/Placebo, AMI/S-MenSCs, and AMI/S-MenSCs*. AMI models were generated using a closed chest coronary occlusion-reperfusion procedure and, after 72 h, the different treatments were intrapericardially administered. Cardiac function parameters were monitored by magnetic resonance imaging before and 7 days post-therapy. Transcriptomic analyses in the infarcted tissue identified 571 transcripts associated with the Gene Ontology term Immune response, of which 57 were differentially expressed when different biogroups were compared. Moreover, a prediction of the interactions between differentially expressed genes (DEGs) and miRNAs from secretomes revealed that some DEGs in the infarction area, such as STAT3, IGFR1, or BCL6 could be targeted by previously identified miRNAs in secretomes from MenSCs. In conclusion, the intrapericardial administration of secretome early after infarction has a significant impact on the expression of immune-related genes in the infarcted myocardium. This confirms the immunomodulatory potential of intrapericardially delivered secretomes and opens new therapeutic perspectives in myocardial infarction treatment.This study was supported by competitive grants, such as: “PFIS” contract (FI19/00041) from the National Institute of Health Carlos III (ISCIII, 2019 Call Strategic Action in Health 2019) to M.Á.d.P.; Santander Bank “Convenio de colaboración empresarial en actividades de interés general” to F.M.; “Sara Borrell” grant (CD19/00048) from ISCIII to E.L.; grant “TE-0001-19” from Consejería de Educación y Empleo (co-funded by European Social Fund -ESF- “Investing in your future”), ayuda para el fomento de la contratación de personal de apoyo a la investigación en la Comunidad Autónoma de Extremadura to M.P. Costs for experimental development were funded by grant “CB16/11/00494” from CIBER-CV ISCIII, RD21/0017/0014 from ISCIII (co-funded by NextGenerationEU. Plan de Recuperación Transformación y Resiliencia) and Ayuda Grupos catalogados de la Junta de Extremadura (GR21201) from Junta de Extremadura, Consejería de Economía, Ciencia y Agenda Digital (co-funded by European Regional Development Fund—ERDF) to F.M.S.-M.; J.G.C. received fundings from the ISCIII through a “Miguel Servet I” grant (MS17/00021) co-funded by ERDF/ESF “A way to make Europe” “Investing in your future”, funding from the projects “CP17/00021” and “PI18/0911” (co-funded by ERDF/ESF), and by Junta de Extremadura. V.C. received fundings from ISCIII (grant number “PI16/01172” and “PI20/00247”). E.L. received fundings from Junta de Extremadura through a “IB20184” grant (co-funded by ERDF/ESF). The funders had no role in study designs, data collection and analysis, decision to publish, or preparation of the manuscript.S

A fibrin coating method of polypropylene meshes enables the adhesion of menstrual blood-derived mesenchymal stromal cells: a new delivery strategy for stem cell-based therapies

Polypropylene (PP) mesh is well-known as a gold standard of all prosthetic materials of choice for the reinforcement of soft tissues in case of hernia, organ prolapse, and urinary incontinence. The adverse effects that follow surgical mesh implantation remain an unmet medical challenge. Herein, it is outlined a new approach to allow viability and adhesion of human menstrual blood-derived mesenchymal stromal cells (MenSCs) on PP surgical meshes. A multilayered fibrin coating, based on fibrinogen and thrombin from a commercial fibrin sealant, was optimized to guarantee a homogeneous and stratified film on PP mesh. MenSCs were seeded on the optimized fibrin-coated meshes and their adhesion, viability, phenotype, gene expression, and immunomodulatory capacity were fully evaluated. This coating guaranteed MenSC viability, adhesion and did not trigger any change in their stemness and inflammatory profile. Additionally, MenSCs seeded on fibrin-coated meshes significantly decreased CD4+ and CD8+ T cell proliferation, compared to in vitro stimulated lymphocytes (p < 0.0001). Hence, the proposed fibrin coating for PP surgical meshes may allow the local administration of stromal cells and the reduction of the exacerbated inflammatory response following mesh implantation surgery. Reproducible and easy to adapt to other cell types, this method undoubtedly requires a multidisciplinary and translational approach to be improved for future clinical uses.This work was supported by: SANTANDER BANK: “Convenio de colaboración empresarial en actividades de interés general” to F.M.; FUNDAÇÃO PARA A CIÊNCIA E A TECNOLOGIA (FCT): post-doctoral contract CEECIND/01026/2018 to J.M.S.; INSTITUTO DE SALUD CARLOS III (ISCIII): a “PFIS” contract (FI19/00041) to M.Á.P., a “Sara Borrell” grant (CD19/00048) to E.L.; a “Miguel Servet I” grant (MS17/00021), co-funded by the European Social Fund (ESF) “Investing in your future”, and projects CP17/00021 and PI18/0911, co-funded by the European Regional Development Fund (ERDF) “A way to make Europe” to J.G.C.; a “CIBERCV” grant (CB16/11/00494), co-funded by the ERDF to F.M.S.-M; JUNTA DE EXTREMADURA, CONSEJERÍA DE ECONOMÍA, CIENCIA Y AGENDA DIGITAL: project IB20184 (co-funded by ERDF) to E.L. and M.P.; grant GR18199, co-funded by the ERDF, to F.M.S.-M.; contracts TA18054 to I.J. and TA18011 to J.J.L. (cofinanced by FEDER)

Terapias avanzadas basadas en células madre y vesículas extracelulares: caracterización y aplicaciones en modelos animales

Gracias a su capacidad de diferenciación, sus efectos paracrinos y su multipotencialidad, las células madre mesenquimales (MSCs) se han convertido en una prometedora herramienta terapéutica en el campo de la biomedicina. A pesar de ello, este tipo de terapias tiene sus limitaciones, principalmente relacionadas con la seguridad. Por este motivo, la investigación se ha centrado en el uso de terapias basadas en la administración de vesículas extracelulares, las cuales son secretadas por las MSCs para ejercer su efecto paracrino. En la actualidad, se están llevando a cabo numerosos estudios clínicos y preclínicos usando células madre como terapia de diversas enfermedades, algunos de los cuales ya han sido aprobados por la Agencia Europea del Medicamento y por la American Food and Drug administration. Por el contrario, existen muy pocos ensayos clínicos basados en la administración de vesículas extracelulares, ya que este tipo de terapias presentan muchas desventajas y su aplicación clínica sigue siendo un tema controvertido. Por esta razón, el objetivo de esta tesis ha sido caracterizar y evaluar el efecto de las terapias basadas en la administración de MSCs y de sus vesículas extracelulares sobre modelos animales. Para la evaluación de ambas terapias se utilizaron tres condiciones patológicas/fisiológicas, cuya aplicación clínica aún no se ha desarrollado. Así, los modelos animales de hernia abdominal, infarto de miocardio y estudios de fecundación in vitro nos han permitido dilucidar si estas opciones terapéuticas avanzadas pueden ser útiles para prevenir y tratar enfermedades humanas, mejorando y prolongando la calidad de vida.Stem cell therapies have represented one of the brightest promises in the biomedical science in the last twenty years. Mesenchymal stem cells (MSCs) have been broadly studied and their extensive use in regenerative medicine can be attributed to their differentiation ability, multipotentiality, and paracrine potential. Despite this, stem cell therapies have some limitations and safety concerns. Because of that, researchers have focused their attention on cell-free therapies that involve extracellular vesicles (EVs) secreted by MSCs, which are responsible of most of their paracrine activity. Currently, cell-based therapies are undergoing numerous preclinical and clinical trials, having, some of them, already been approved as advanced therapies by the European Medical Agency and by the American Food and Drug administration. In contrast, very little ongoing clinical trials are testing the clinical use of cellfree therapies based on extracellular vesicles. Both therapies still present some disadvantages, and their clinical application is still a controversial issue. For this reason, the aim of this thesis was the characterization and evaluation of MSC- and EV-based therapies in animal models. Three pathological/physiological conditions, whose resolutive clinical approach has not been defined yet, will be used for the evaluation of these cell- and cell-free therapies. Hence, animal models of abdominal hernia, myocardial infarction and in vitro fecundation studies allowed us to elucidate if these advanced therapeutic options may be useful to prevent and treat human diseases, improving and prolonging quality of life.This thesis was supported by a MAFRESA S.L. grant (part of Grupo Jorge, promoted by Jesús Usón Gargallo) to Federica Marinaro; “Sara Borrell” grant (CD19/00048) from the National Institute of Health Carlos III (ISCIII) to Esther López Nieto; Miguel Servet I” grant “MS17/00021” [co-funded by ERDF/European Social Fund (ESF) “A way to make Europe”/“Investing in your future”] and projects “CP17/00021” and “PI18/0911” (co-funded by ERDF/ESF) from ISCIII and “IB16168” grant (cofunded by ERDF/ESF) by Junta de Extremadura to Javier García Casado; grant “CB16/11/00494” from CIBER-CV and Ayuda Grupos de Investigación de Extremadura (GR18199) from Consejería de Economía, Ciencia y Agenda Digital (cofunded by European Regional Development Fund – ERDF) to Francisco Miguel Sánchez Margallo and by Jesús Usón Minimally Invasive Surgery Centre (CCMIJU). This thesis has been developed at the ICTS Nanbiosis (Unit 14 — Stem Cell Therapy, Unit 21 — Experimental Operating Rooms, Unit 22 — Animal Housing, Unit 23 — Assisted Reproduction, Unit 24 — Medical Imaging) in collaboration with CNIC (Centro Nacional de Investigaciones Cardiovasculares, Spain), INIA (Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria), University of Extremadura (Spain), University of Marburg (Germany), University of Porto (Portugal), and Federal University of Santa Maria (Brazil). The funders had no role in study designs, data collection and analysis, decision to publish, or preparation of the manuscripts collected in this thesis

Modulation of gonadotrophin induced steroidogenic enzymes in granulosa cells by d-chiroinositol

d-chiroinositol (DCI) is a inositolphosphoglycan (IPG) involved in several cellular functions that control the glucose metabolism. DCI functions as second messenger in the insulin signaling pathway and it is considered an insulin sensitizer since deficiency in tissue availability of DCI were shown to cause insulin resistance (IR). Polycystic ovary syndrome (PCOS) is a pathological condition that is often accompanied with insulin resistance. DCI can positively affects several aspect of PCOS etiology decreasing the total and free testosterone, lowering blood pressure, improving the glucose metabolism and increasing the ovulation frequency. The purpose of this study was to evaluate the effects of DCI and insulin combined with gonadotrophins namely follicle-stimulating hormone (FSH) and luteinizing hormone (LH) on key steroidogenic enzymes genes regulation, cytochrome P450 family 19 subfamily A member 1 (CYP19A1) and cytochrome P450 side-chain cleavage (P450scc) in primary cultures of human granulosa cells (hGCs). We also investigated whether DCI, being an insulin-sensitizer would be able to counteract the expected stimulator activity of insulin on human granulosa cells (hGCs). Methods. The study was conducted on primary cultures of hGCs. Gene expression was evaluated by RT-qPCR method. Statistical analysis was performed applying student t-test, as appropriate (P < 0.05) set for statistical significance. Results. DCI is able to reduce the gene expression of CYP19A1, P450scc and insulin-like growth factor 1 receptor (IGF-1R) in dose–response manner. The presence of DCI impaired the increased expression of steroidogenic enzyme genes generated by the insulin treatment in gonadotrophin-stimulated hGCs. Conclusions. Insulin acts as co-gonadotrophin increasing the expression of steroidogenic enzymes genes in gonadotrophin-stimulated granulosa cells. DCI is an insulin-sensitizer that counteracts this action by reducing the expression of the genes CYP19A1, P450scc and IGF-1R. The ability of DCI to modulate in vitro ovarian activity of insulin could in part explain its beneficial effect when used as treatment for conditions associated to insulin resistance

Модуляція індукованих гонадотропіном стероїдогенних ферментів в клітинах гранульози за допомогою D-хіро-інозитолу

d-chiroinositol (DCI) is a inositolphosphoglycan (IPG) involved in several cellular functions that control the glucose metabolism. DCI functions as second messenger in the insulin signaling pathway and it is considered an insulin sensitizer since deficiency in tissue availability of DCI were shown to cause insulin resistance (IR). Polycystic ovary syndrome (PCOS) is a pathological condition that is often accompanied with insulin resistance. DCI can positively affects several aspect of PCOS etiology decreasing the total and free testosterone, lowering blood pressure, improving the glucose metabolism and increasing the ovulation frequency.The purpose of this study was to evaluate the effects of DCI and insulin combined with gonadotrophins namely follicle-stimulating hormone (FSH) and luteinizing hormone (LH) on key steroidogenic enzymes genes regulation, cytochrome P450 family 19 subfamily A member 1 (CYP19A1) and cytochrome P450 side-chain cleavage (P450scc) in primary cultures of human granulosa cells (hGCs). We also investigated whether DCI, being an insulin-sensitizer would be able to counteract the expected stimulator activity of insulin on human granulosa cells (hGCs).Methods. The study was conducted on primary cultures of hGCs. Gene expression was evaluated by RT-qPCR method. Statistical analysis was performed applying student t-test, as appropriate (P &lt; 0.05) set for statistical significance.Results. DCI is able to reduce the gene expression of CYP19A1, P450scc and insulin-like growth factor 1 receptor (IGF-1R) in dose–response manner. The presence of DCI impaired the increased expression of steroidogenic enzyme genes generated by the insulin treatment in gonadotrophin-stimulated hGCs.Conclusions. Insulin acts as co-gonadotrophin increasing the expression of steroidogenic enzymes genes in gonadotrophin-stimulated granulosa cells. DCI is an insulin-sensitizer that counteracts this action by reducing the expression of the genes CYP19A1, P450scc and IGF-1R. The ability of DCI to modulate in vitro ovarian activity of insulin could in part explain its beneficial effect when used as treatment for conditions associated to insulin resistance.D-хиро-инозитол (DCI) - это инозитолфосфогликан, который включает в себя несколько клеточных функций, контролирующих метаболизм глюкозы. DCI функционирует как второй посредник для сигнализации действия инсулина и считается сенсибилизатором инсулина, поскольку дефицит наличия DCI в тканях указывает на инсулинорезистентность (ИР).Синдром поликистозных яичников (СПКЯ) – патологическое состояние, которое часто сопровождается ИР. DCI положительно влияет на многочисленные аспекты этиологии СПКЯ, уменьшает общий и свободный уровень тестостерона, снижает кровяное давление, нормализует метаболизм глюкозы и увеличивает частоту овуляции.Цель этого исследования заключалась в оценке влияния DCI и инсулина в сочетании с гонадотропинами, а именно фолликулостимулирующим и лютеинизирующим гормонами на регуляцию генов стероидогенных ферментов, подсемейств A1 (CYP19A1) и расщепления боковых цепей цитохрома P450 (P450scc) семейства цитохрома P450 в первичных культурах клеток гранулезы человека. Мы также исследовали, сможет ли DCI как инсулиносенситайзер противодействовать ожидаемой активности инсулина стимулировать клетки гранулезы человека.Методы. Исследование проводилось на основных культурах клеток гранулезы человека. Экспрессию генов оценивали по методу RT-qPCR. Статистический анализ проводился с использованием t-критерия Стьюдента (p &lt; 0,05), установленного для статистической значимости.Результаты. DCI способен уменьшать экспрессию генов CYP19A1, P450scc и рецептора инсулиноподобного фактора роста 1 с дозозависимым ответом. Наличие DCI сопровождается усилением экспрессии генов стероидогенных ферментов, производимых гонадотропин-стимулированными клетками гранулезы человека при лечении инсулином.Выводы. Инсулин выступает в роли ко-гонадотропина, что увеличивает экспрессию генов стероидогенных ферментов в клетках гранулезы, стимулированных гонадотропином. DCI – это сенсибилизатор инсулина, который противодействует этому влиянию, уменьшая экспрессию генов CYP19A1, P450scc и рецептора инсулиноподобного фактора роста 1. Способность DCI модулировать активность инсулина в яичнике in vitro может частично объяснить его полезный эффект, когда он используется для лечения состояний, связанных с ИР.D-хіро-інозитол (DCI) – це інозитолфосфоглікан, який включає в себе кілька клітинних функцій, що контролюють метаболізм глюкози. DCI функціонує як другий посередник для сигналізації дії інсуліну і вважається сенсибілізатором інсуліну, оскільки дефіцит наявності DCI у тканинах вказує на інсулінорезистентність (ІР).Синдром полікістозних яєчників (СПКЯ) – патологічний стан, який часто супроводжується ІР. DCI позитивно впливає на численні аспекти етіології СПКЯ, зменшує загальний та вільний рівень тестостерону, знижує кров’яний тиск, нормалізує метаболізм глюкози та збільшує частоту овуляції.Мета цього дослідження полягала в оцінці впливу DCI та інсуліну в поєднанні з гонадотропінами, а саме фолікулостимулюючим та лютеїнізуючим гормонами на регуляцію генів стероїдогенних ферментів, підродини A1 (CYP19A1) та розщеплення бічних ланцюгів цитохрому P450 (P450scc) родини цитохрому P450 у первинних культурах клітин гранульози людини. Ми також досліджували, чи зможе DCI як інсуліносенсітайзер протидіяти очікуваній активності інсуліну стимулювати клітини гранульози людини.Методи. Дослідження проводилося на основних культурах клітин гранульози людини. Експресію генів оцінювали методом RT-qPCR. Статистичний аналіз проводився з використанням t-критерію Стьюдента (p &lt; 0,05), встановленого для статистичної значущості.Результати. DCI здатний зменшувати експресію генів CYP19A1, P450scc та рецептора інсуліноподібного фактора росту 1 у дозозалежній відповіді. Наявність DCI супроводжується посиленням експресії генів стероїдогенних ферментів, вироблених гонадотропін-стимульованими клітинами гранульози людини при лікуванні інсуліном.Висновки. Інсулін виступає в ролі ко-гонадотропіну, що збільшує експресію генів стероїдогенних ферментів у клітинах гранульози, стимульованих гонадотропіном. DCI – це сенсибілізатор інсуліну, який протидіє цьому впливу, зменшуючи експресію генів CYP19A1, P450scc та рецептора інсуліноподібного фактора росту 1. Здатність DCI модулювати активність інсуліну в яєчнику in vitro може частково пояснити його корисний ефект, коли він використовується для лікування станів, пов’язаних із ІР