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OBJECTIVE: To evaluate the effectiveness and safety of Zhenqi Buxue Oral Liquid (ZQ), progesterone capsules, and their combination in treating oligomenorrhea and hypomenorrhea with qi-blood and Kidney (Shen) essence deficiency. METHODS: This was a prospective, randomized, multi-center controlled trial between June 2022 to December 2022. Ninety-six oligomenorrhea and hypomenorrhea patients with qi-blood and Shen essence deficiency were randomly assigned to receive ZQ (ZQ group, 29 cases), progesterone capsules (PG group, 32 cases), or the combined Chinese and Western medicine (COM group, 31 cases) at a ratio of 1:1:1. Patients in the ZQ or PG group took daily 10 mL twice a day of ZQ or 200 mg once a day of progesterone capsules for 10 consecutive days on day 15 of the menstrual cycle respectively, and patients in the COM group received the same ZQ combined with progesterone capsules. The treatment course lasted for 3 months and follow-up was performed at 1 and 3 months after the end of treatment. Primary endpoint was the menstrual Traditional Chinese Medicine Syndrome Scale (TCMSS) scores. Secondary endpoints included pictorial blood loss assessment chart (PBAC) scores, clinical efficacy rate, 36-item Short Form Health Survey (SF-36) scores, sex hormones and thickness of endometrium. Adverse events (AEs) were recorded. RESULTS: TCMSS scores after 1- and 3-month treatment in all groups were significantly lower than those at baseline (P<0.05). Only TCMSS scores after 3-month treatment in the ZQ and COM groups continuously decreased compared with those after 1-month treatment in the same group (P<0.01). TCMSS scores after 3-month treatment in the ZQ and COM groups were significantly lower than those in the PG group (P<0.05, P<0.01). Compared with baseline, PBAC scores in the ZQ and COM groups after 3 months of treatment were also significantly higher (both P<0.01). The total effective rates of TCM syndrome of 3-month treatment were significantly improved in all groups compared with that after 1 month of treatment (P<0.05). The total effective rate of the COM group was the highest in the 3rd month of treatment and significantly higher than that of PG group alone (P<0.05). Compared with baseline, only the SF-36 scores of COM group were significantly improved after 3 months of treatment (P<0.05). No serious adverse reactions were observed after treatment. CONCLUSIONS: The combination of ZQ and PG, or ZQ only had better effects on reducing TCMSS scores compared with PG, and COM showed the higher total effective rate compared with monotherapy. Besides, COM could effectively improve menstrual blood loss and quality of life. ZQ combined with PG may be an effective and safe option for oligomenorrhea and hypomenorrhea patients with qi-blood and Shen essence deficiency.
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Medicamentos Herbarios Chinos , Progesterona , Femenino , Humanos , Progesterona/uso terapéutico , Qi , Oligomenorrea/tratamiento farmacológico , Calidad de Vida , Estudios Prospectivos , Medicina Tradicional China , Medicamentos Herbarios Chinos/efectos adversos , Cápsulas , RiñónRESUMEN
BACKGROUND: Our study sought to obtain data which assess the diagnostic value of transvaginal three-dimensional ultrasound (3D-US) combined with color Doppler ultrasound (US) for early cesarean scar pregnancy (CSP). METHODS: All participants were randomly divided into a Control group diagnosed using 3D-US and a Combination group diagnosed using 3D-US combined with color Doppler US. The preoperative US results were compared with postoperative pathological results. The diagnostic coincidence rate, sensitivity, and specificity of these two examination methods were compared, and their diagnostic results for different types of CSP were analyzed. Finally, the diagnostic effects of both methods were compared and analyzed, and the imaging of CSP was summarized. RESULTS: The diagnostic accuracy of transvaginal 3D-US combined with color Doppler US (92.96%) was significantly higher than that of transvaginal 3D-US (71.83%). For different types of CSP, the diagnostic rate of CSP with mixed echogenic mass and partial implantation of gestational sac in the Combination group was markedly higher than that in the Control group. CONCLUSIONS: Additionally, the sensitivity, specificity, and area under the receiver operating characteristic (ROC) curve (AUC) in the Combination group were higher than those in the Control group. Transvaginal 3D-US combined with color Doppler US can improve the sensitivity, specificity, and accuracy of diagnosis of early CSP, and has important reference value for clinical condition evaluation and treatment options.
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Cicatriz , Embarazo Ectópico , Cesárea/efectos adversos , Cicatriz/diagnóstico por imagen , Femenino , Humanos , Embarazo , Embarazo Ectópico/diagnóstico por imagen , Ultrasonografía , Ultrasonografía Doppler en ColorRESUMEN
Polycystic ovary syndrome (PCOS), one of the most common types of endocrine diseases, is characterized by a high prevalence among women of reproductive-age. However, its pathogenesis and molecular mechanisms remain unclear. CircMTO1 has been reported to participate in numerous biological processes, but, its role in PCOS progression remains unknown. In the current study, we elucidated the expression and circRNA characterization of circMTO1 in human granulosa-like tumor cells. We found that circMTO1 knockdown promoted human granulosa-like tumor cell proliferation and inhibited its apoptosis rate. Next, we explored the underlying molecular mechanisms by using a series of experiments. Our results revealed the effect of the novel circMTO1/miR-320b/MCL1 axis in human granulosa-like tumor cells. Furthermore, we found that the expression of circMTO1 was induced by Snail family transcriptional repressor 2 (SNAI2) in human granulosa-like tumor cells. Our results may provide potential targets for PCOS research and a novel direction for the diagnosis and treatment of PCOS.
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RATIONAL: Ovarian sertoli-leydig cell tumor (OSLCT) is extremely rare. We reported a OSLCT case in whom postmenopausal vaginal bleeding was the first symptom. PATIENT CONCERNS: The patient came to our hospital due to postmenopausal vaginal bleeding. DIAGNOSES: Serum tumor markers and color Doppler ultrasound for her pelvic cavity were negative. The patient was finally diagnosed with left OSLCT by pathology. It was difficult to make a definite diagnosis before operation, the diagnosis of OSLCT required postoperative pathology in the patients. INTERVENTIONS: the patient underwent laparoscopic hysterectomy+bilateral adnexectomy+lysis of pelvic adhesions. OUTCOMES: Postoperative laboratory examinations were normal. The patient was discharged from our hospital on the seventh day after operation and came to our hospital for follow-up check in April 2020. Physical and laboratory examinations were normal. LESSONS: OSLCT can show different endocrine abnormalities, which are related to the various types of tumor tissues. Missed diagnosis and misdiagnosis are likely to occur in the patients who only have elevated serum testosterone. For the menopausal women with elevated serum testosterone, ovarian tumor shoule be highly suspected after excluding adrenal gland-related diseases.
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Neoplasias Ováricas/complicaciones , Tumor de Células de Sertoli-Leydig/complicaciones , Hemorragia Uterina/etiología , Femenino , Humanos , Histerectomía , Persona de Mediana Edad , Neoplasias Ováricas/cirugía , Posmenopausia , Tumor de Células de Sertoli-Leydig/cirugía , Hemorragia Uterina/cirugíaRESUMEN
INTRODUCTION: Ovarian cancer is one of the leading causes of cancer-related deaths in women. Treatments for ovarian cancer include surgery followed by chemotherapy. However, the survival rate for ovarian cancer is still not satisfactory. Moreover, the current chemotherapy has numerous associated side effects. Therefore there is an urgent need to look for novel and more viable treatment options. Against this backdrop the present study was designed to evaluate the anticancer activity of sugiol against ovarian cancer cells. MATERIAL AND METHODS: Cell viability was assessed by CCK8 assay, apoptosis by DAPI, AO/ER and annexin V/PI staining. Mitochondrial membrane potential and cell cycle analysis was performed by flow cytometry. Cell migration was investigated by wound healing assay. Protein expression was monitored by western blotting. RESULTS: The results of the present study indicated that sugiol exerts significant (p < 0.0001) anticancer effects on SKOV3 cancer cells with an IC50 of 25 µM. However, sugiol exhibited less cytotoxicity against normal ovarian cells with an IC50 of 62.5 µM. The anticancer effects of sugiol were found to be due to G0/G1 cell cycle arrest and mitochondrial apoptosis. Sugiol also inhibited cell migration of SKOV3 cells dose dependently. Moreover, the results showed that sugiol could inhibit the RAF/MEK/ERK signalling pathway in a dose-dependent manner. CONCLUSIONS: The results of the present study indicate that sugiol exerts potent anticancer effects on SKOV3 cells via induction of cell cycle arrest, mitochondrial apoptosis and inhibition of the RAF/MEK/ERK signalling pathway.
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Superovulation procedures and assisted reproductive technologies have been widely used to treat couples who have infertility problems. Although generally safe, the superovulation procedures are associated with a series of complications, such as ovarian hyper-stimulation syndrome, thromboembolism, and adnexal torsion. The role of long-term repeated superovulation in ovarian aging and especially in associated disorders such as osteoporosis and cardiovascular diseases is still unclear. In this study, we sought to determine if repeated superovulation by ten cycles of treatment with pregnant mare serum gonadotropin/human chorionic gonadotropin could affect ovarian reserve, ovarian function, bone density and heart function. Ovarian reserve and function were reflected by the size of the primordial follicle pool, anti-Mullerian hormone expressions, hormone levels and fertility status. Furthermore, we examined bone density and heart function by microCT and cardiovascular ultrasonography, respectively. After repeated superovulation, the size of the primordial follicle pool and the expression of anti-mullerian hormone decreased, along with the concentrations of estrogen and progesterone. Mice exposed to repeated superovulation showed an obvious decrease in fertility and fecundity. Furthermore, both bone density and heart ejection fraction significantly decreased. These results suggest that repeated superovulation may increase the risk of osteoporosis and cardiovascular diseases by accelerating ovarian aging.
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Densidad Ósea/efectos de los fármacos , Reserva Ovárica/efectos de los fármacos , Inducción de la Ovulación/efectos adversos , Volumen Sistólico/efectos de los fármacos , Superovulación , Animales , Enfermedades Cardiovasculares/etiología , Gonadotropina Coriónica/toxicidad , Combinación de Medicamentos , Femenino , Gonadotropinas Equinas/toxicidad , Ratones , Ratones Endogámicos C57BL , Osteoporosis/etiología , Ovario/efectos de los fármacos , Ovario/patología , Inducción de la Ovulación/métodosRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Kuntai capsule, a traditional Chinese medicine, has been widely used for the clinical treatment of menopausal syndrome. However, its mechanisms remain poorly understood. Considering that aging ovaries are the primary cause of menopause, this study was designed to investigate the effects and mechanisms of Kuntai capsule on ovarian function in a novel mice model with accelerated aging ovaries. MATERIALS AND METHODS: Seventy-five female C57BL/6 mice were chosen for this study. Fifteen of the mice were separated into the normal control group (NC). The remaining sixty were used to establish the novel accelerated aging ovary model by superovulation and oxidative stress and then by randomly dividing the mice into four equal groups. One group was considered the model group (Mod). The other three groups were treated with low (0.4g/kg), middle (0.8g/kg) and high (1.6g/kg) doses of Kuntai capsule intragastrically every day for 4 weeks. During the treatment, the body weight and fur condition of all mice were recorded. All the mice were forced to swim to record their exhaustive swimming time (EST), which measures their strength. Mice were then sacrificed for sampling. Ovarian reserve was evaluated using follicle counts and AMH expression. Ovarian function was evaluated using estrous cycle, sex hormone level and litter experiments. Ovarian follicles were categorized and counted to estimate ovarian reserve, and ovarian histologic sections were stained for terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) to detect apoptotic cells. The ultrastructure of ovarian cells was observed using transmission electron microscopy. Western blotting was used to measure expression of Bax, Bcl2, AMH and SOD2 protein. RESULTS: Compared with the NC GROUP, the Mod group clearly displayed worse fur condition and ovarian function. These situations showed some improvement after Kuntai capsule treatment. Specifically, the fur condition and the EST of the Kuntai capsule groups were superior to the fur condition and EST of the Mod group. In cases of damaged ovarian function, Kuntai capsule can regulate the estrous cycles, increase hormone secretion and fertility and significantly decrease atretic follicles. The transmission electron microscopy results revealed that Kuntai capsule rescued the ovarian ultrastructure of mice. TUNEL staining confirmed that the apoptotic cells were reduced after Kuntai capsule treatment. Western blotting revealed that Kuntai capsule can increase AMH, SOD2, and Bcl2 protein expression and decrease Bax expression. CONCLUSIONS: Kuntai capsule may improve damaged ovarian function, which may be related to its antioxidant and anti-apoptosis effects.
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Antioxidantes/farmacología , Medicamentos Herbarios Chinos/farmacología , Reserva Ovárica/efectos de los fármacos , Ovario/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Administración Oral , Factores de Edad , Animales , Hormona Antimülleriana/metabolismo , Antioxidantes/administración & dosificación , Apoptosis/efectos de los fármacos , Cápsulas , Citoprotección , Relación Dosis-Respuesta a Droga , Medicamentos Herbarios Chinos/administración & dosificación , Ciclo Estral/efectos de los fármacos , Femenino , Fertilidad/efectos de los fármacos , Exposición por Inhalación , Ratones Endogámicos C57BL , Fuerza Muscular , Tamaño de los Órganos , Folículo Ovárico/efectos de los fármacos , Folículo Ovárico/ultraestructura , Ovario/metabolismo , Ovario/fisiopatología , Ovario/ultraestructura , Ozono/toxicidad , Embarazo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Superovulación , Superóxido Dismutasa/metabolismo , Factores de Tiempo , Proteína X Asociada a bcl-2/metabolismoRESUMEN
Ovarian aging is a long-term and complex process associated with a decrease in follicular quantity and quality. The damaging effects of reactive oxygen species (ROS) in ovarian aging and ovarian aging-associated disorders have received relatively little attention. Thus, we assessed if the oxidative stress induced by long-term (defined by the Environmental Protection Agency as at least 30 days in duration) moderate ozone inhalation reduced ovarian reserves, decreased ovarian function and induced ovarian aging-associated disorders. The expression of oxidative stress markers and antioxidant enzymes was used to determine the degree of oxidative stress. Ultrastructural changes in ovarian cells were examined via electron microscopy. The ovarian reserve was assessed by measuring multiple parameters, such as the size of the primordial follicle pool and anti-Müllerian hormone (AMH) expression. The estrous cycle, hormone levels and fertility status were investigated to assess ovarian function. To investigate ovarian aging-associated disorders, we utilized bone density and cardiovascular ultrasonography in mice. The levels of oxidized metabolites, such as 8-hydroxy-2´-deoxyguanosine (8-OHdG), 4-hydroxynonenal (4-HNE) and nitrotyrosine (NTY), significantly increased in ovarian cells in response to increased oxidative stress. The ultrastructural analysis indicated that lipid droplet formation and the proportion of mitochondria with damaged membranes in granulosa cells were markedly increased in ozone-exposed mice when compared with the control group. Ozone exposure did not change the size of the primordial follicle pool or anti-Müllerian hormone (AMH) expression. The estrogen concentration remained normal; however, progesterone and testosterone levels decreased. The mice exposed to ozone inhalation exhibited a substantial decrease in fertility and fecundity. No differences were revealed by the bone density or cardiovascular ultrasounds. These findings suggest that the decreased female reproductive function caused by long-term moderate oxidative damage may be due to a decrease in follicle quality and progesterone production.
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Numerous studies have reported that human umbilical cord mesenchymal stem cell (hUCMSC) therapy can rescue the structure and function of injured tissues. The aims of this study were to explore the protective role of hUCMSC transplantation in a model of accelerated ovarian aging and to compare 2 methods of transplanting hUCMSCs, i.e. i) via intravenous injection (IV) and ii) in situ ovarian micro injection (MI). Female mice were subjected to superovulation and ozone inhalation to create a model of accelerated ovarian aging with a decline in both the quantity and quality of oocytes. Cells were transplanted via IV or MI, and ovaries were removed after 2 weeks or 1 month of treatment. Ovarian reserve and function were evaluated based on the follicle counts, hormone levels, the estrous cycle, and reproductive performance. Cell tracking, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), real-time polymerase chain reaction (PCR), and Western blot analysis were used to assess the inner mechanisms of injury and repair. Results indicated that ovarian function increased significantly after treatment with hUCMSCs. Immunofluorescence revealed reduced TUNEL staining and a decreased percentage of apoptotic cells. A higher level of expression of anti-apoptotic and antioxidant enzymes was noted in the ovaries of groups treated with hUCMSCs. These parameters were enhanced more when mice were treated with hUCMSCs for 1 month than when they treated with hUCMSCs for 2 weeks. IV was better able to restore ovarian function than MI. These results suggest that both methods of transplantation may improve ovarian function and that IV transplantation of hUCMSCs can significantly improve ovarian function and structural parameters more than MI transplantation of hUCMSCs can.
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Tratamiento Basado en Trasplante de Células y Tejidos , Trasplante de Células Madre Mesenquimatosas/métodos , Enfermedades del Ovario/terapia , Cordón Umbilical/citología , Animales , Antioxidantes/metabolismo , Apoptosis/genética , Rastreo Celular , Ciclo Estral , Femenino , Regulación de la Expresión Génica , Humanos , Etiquetado Corte-Fin in Situ , Ratones , Ratones Endogámicos C57BL , Oocitos/patología , Reserva Ovárica , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Sirtuins, a family of nicotinamide adenine dinucleotide (NAD)-dependent deacetylases that play diverse roles in regulating metabolism, cell proliferation, and genome stability, have been implicated in mammalian aging and age-related diseases, including cancers, metabolic disorders, and neurodegenerative diseases. Ovarian aging is thought to be characterized by a gradual decrease in both the number of follicles and the quality of oocytes. Ovarian reserve is indicated by the number of primordial follicles. In this study, ovarian reserve was assessed in mice of different ages and mice subjected to caloric restriction (CR) and chemotherapy (2 commonly used models for ovarian aging research) by counting primordial follicles and determining the expression levels of SIRT1, SIRT3, and SIRT6 to explore the relationship between ovarian function and sirtuin expression. A gradual decline in the number of follicles (especially primordial follicles) was observed in aging mice and mice subjected to chemotherapy. Histological analysis showed that CR mice displayed a significantly greater number of primordial follicles and less atretic follicles. Western blot analysis indicated that expression levels of SIRT1, SIRT3, and SIRT6 were significantly decreased in the ovaries of aged mice and mice treated with chemotherapy, but increased in CR mice. SIRT1, SIRT3, and SIRT6 all showed a significantly positive correlation with the numbers of primordial follicles (r(2)=0.6399, P<0.0001; r(2)=0.5445, P<0.001; and r(2)=0.4956, P<0.0001, respectively). These results indicate that SIRT1, SIRT3 and SIRT6 are closely related to ovarian reserve, and suggest that these sirtuins may be markers of ovarian aging.
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Biomarcadores/metabolismo , Ovario/citología , Sirtuina 1/metabolismo , Sirtuina 3/metabolismo , Sirtuinas/metabolismo , Animales , Restricción Calórica , Recuento de Células , Senescencia Celular , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/metabolismo , Femenino , Regulación de la Expresión Génica , Ratones , Folículo Ovárico/citología , Ovario/fisiologíaRESUMEN
Theca-interstitial cells (TICs) and granulosa cells (GCs) are important components of follicles that support follicle development and hormone secretion, and are considered to be important cell models for basic research. However, no method currently exists for simultaneously isolating TICs and GCs from a single ovary of the immature mouse. Here, we sought to develop such a protocol using mechanical dissection combined with brief collagenase-DNase digestion. Morphological characteristics and molecular markers were detected to identify TICs and GCs. In isolated TICs, cholesterol side chain cleavage cytochrome P450 (P450scc) was expressed abundantly, but anti-Mullerian hormone (AMH) was expressed only at very low levels. This expression profile was reversed in GCs. In addition, TICs secreted large amounts of testosterone (T) and minimal amounts of estradiol (E2 ), while the converse was found in GCs. T concentrations rose gradually in TIC culture media as the concentration of added luteinizing hormone (LH) was increased. In GCs, E2 secretion increased as the follicle-stimulating hormone (FSH) concentration increased. Thus, mechanical dissection combined with collagenase-DNase digestion is a simple, effective and reproducible method for obtaining large numbers of highly purified and hormonally stimulated TICs and GCs from one ovary.
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Células de la Granulosa/citología , Maduración Sexual , Células Tecales/citología , Animales , Hormona Antimülleriana/metabolismo , Separación Celular , Estradiol/metabolismo , Femenino , Hormona Folículo Estimulante/metabolismo , Células de la Granulosa/metabolismo , Hormona Luteinizante/metabolismo , Ratones , Ratones Endogámicos C57BL , Maduración Sexual/genética , Células Tecales/metabolismo , TranscriptomaRESUMEN
The initiation of primordial follicle development is essential for female fertility, but the signals that trigger this process are poorly understood. Given the potentially important roles of microRNAs (miRNAs) in the ovary, we aimed to study the expression patterns and regulatory functions of miRNAs during the initiation of primordial follicle development. Expression patterns of miRNA in the neonatal mouse ovary were profiled by microarray, and 24 miRNAs whose abundances differed significantly between ovaries from 3- and 5-day-old mice were identified. Pathway enrichment analysis revealed that 48 signal transduction pathways are modulated by the up-regulated miRNAs and 29 pathways are modulated by the down-regulated miRNAs (P-value and false discovery rate < 0.001). A miRNA-mRNA regulatory network was established for TGF-beta signaling pathway-related genes. Among the miRNAs involved in this pathway, miR-145 was chosen for further analysis. Down-regulation of miR-145 using an antagomir (AT) decreased the proportion and number of the primordial follicles and increased that of the growing follicles in the cultured ovaries (P < 0.05). The mean oocyte diameter in the primordial follicles was significantly greater in the AT group relative to the AT-negative control group (P < 0.05), whereas the mean oocyte diameter in growing follicles was smaller in the AT group than in the AT-negative control group. In addition, we confirmed that miR-145 targets Tgfbr2. The miR-145 AT caused an increase in TGFBR2 expression and activation of Smad signaling but did not affect the p38 MAPK or JNK pathway. These data suggest that miRNAs and the signaling pathways they modulate are involved in the initiation of primordial follicle development, and miR-145 targets Tgfbr2 to regulate the initiation of primordial follicle development and maintain primordial follicle quiescence.
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Regulación del Desarrollo de la Expresión Génica , Redes Reguladoras de Genes , MicroARNs/genética , Folículo Ovárico/crecimiento & desarrollo , Animales , Animales Recién Nacidos , Diferenciación Celular/genética , Femenino , Perfilación de la Expresión Génica , Ratones , Ratones Endogámicos C57BL , Análisis por Micromatrices , Ovario/citología , Ovario/crecimiento & desarrolloRESUMEN
Mouse models have been widely utilized to elucidate the basic principles and regulatory mechanisms of primordial follicle activation. Outside their natural environment, the growth of follicles might be affected by unknown factors in vitro and the elimination of regulation in vivo. Currently, in vitro culture and transplantation of ovaries under the kidney capsule are two commonly used incubation methods. However, the limited number of studies that have been published compare various incubation systems and reveal differences between ovaries that are incubated and grown in vivo. We compare the number of primordial, primary and secondary follicles in cultured, transplanted and in-vivo-grown ovaries. We investigate the expression levels of four genes, including zona pellucida 3 (ZP3), growth and differentiation factor-9 (GDF-9), proliferating cell nuclear antigen (PCNA) and anti-Müllerian hormone (AMH). Our results suggest that in vitro culture accelerates follicle activation, delays the transition from primary to secondary follicles and affects the expression patterns of ZP3, GDF-9, PCNA and AMH. A larger number of secondary follicles in ovaries cultured in alpha-minimal essential medium (α-MEM) had intact zona pellucida compared with those grown in Dulbecco's modified Eagle medium containing Ham's F-12 nutrient mixture (D/F12), suggesting that α-MEM is a better basal medium. The transplanted ovaries demonstrated the most similar characteristics to the in-vivo-grown ovaries, indicating that transplantation provided an optimal environment for ovarian incubation. This study has thus established the similarities and differences between in-vivo-grown and incubated ovaries, demonstrated that transplantation can mostly mimic the environment of ovarian growth in vivo and determined the optimal basal culture medium between α-MEM and D/F12.
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Ovario/crecimiento & desarrollo , Ovario/trasplante , Animales , Femenino , Regulación del Desarrollo de la Expresión Génica , Células de la Granulosa/citología , Células de la Granulosa/metabolismo , Células de la Granulosa/ultraestructura , Ratones , Ratones Endogámicos C57BL , Oocitos/citología , Oocitos/metabolismo , Oocitos/ultraestructura , Técnicas de Cultivo de Órganos , Folículo Ovárico/crecimiento & desarrollo , Folículo Ovárico/metabolismo , Folículo Ovárico/ultraestructura , Ovario/metabolismo , Ovario/ultraestructuraRESUMEN
OBJECTIVE: To study the protective effects on ovarian function by caloric restriction (CR) and its mechanism. METHODS: Thirty female C57BL/6 mice of 8 weeks old were randomly divided into two groups, including ad libitum (AL) group and caloric restriction (CR) group. The general situation and ovarian function of those mice were compared and evaluated.Ovarian follicles were counted by hematoxylin-eosin staining. Anti-Miillerian Hormone(AMH) mRNA expression of the ovary were detected by using real-time PCR. The concentrations of serum estradiol, progesterone of the mice were measured by ELISA. And the fertility of mice by mating trials were evaluated, SIRT3, Hypoxia inducible factor 1α (HIF-1α) and catalase (CAT) mRNA expression of the mice ovaries were detected by Real-Time PCR. RESULTS: The total follicles were 546 in CR mice and 286 in AL mice. The proportion of primordial follicles were 38.6% (211/546) in ovaries of CR mice and 29.4% (84/286) in ovaries of AL mice, which reached statistical difference. The proportion of atretic follicles 5.3% (29/546) in ovaries of CR mice, compared with 16.8% (48/286) in AL mice, was significantly decreased (P < 0.05). The AMH mRNA expression in CR mice ovaries was 3.37 times of that of AL mice (P < 0.05). The serum concentration of estradiol in CR mice was up to (5.3 ± 1.6) pmol/L, which was much higher than (3.6 ± 1.6) pmol/L in AL mice. While, the progesterone concentration of (0.4 ± 0.3) nmol/L in CR mice was lower than (1.4 ± 0.8) nmol/L in AL mice (P < 0.05).Fertility and survival of offsprings were both improved in CR mice. The expression level of SIRT3 mRNA in CR mice ovary was 1.39 times, CAT was 1.55 times and HIF-1α was 0.31 times of those in AL mice (P < 0.05). CONCLUSIONS: Caloric restriction can delay the ovary aging process through reduce follicle depletion by suppressing follicle recruitment and ovulation. The function of ovarian reserve and reproductive endocrine was effectively protected. Caloric restriction can reduce the incidence of follicular atresia, its mechanism might be associated with anti-oxidative stress.