RESUMEN
Targeting dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A) has been verified to regulate the progression of tau pathology as a promising treatment for Alzheimer's disease (AD), while the research progress on DYRK1A inhibitors seemed to be in a bottleneck period. In this work, we identified 32 (ZJCK-6-46) as the most potential DYRK1A inhibitor (IC50 = 0.68 nM) through rational design, systematic structural optimization, and comprehensive evaluation. Compound 32 exhibited acceptable in vitro absorption, distribution, metabolism, and excretion (ADME) properties and significantly reduced the expression of p-Tau Thr212 in Tau (P301L) 293T cells and SH-SY5Y cells. Moreover, compound 32 showed favorable bioavailability, blood-brain barrier (BBB) permeability, and the potential of ameliorating cognitive dysfunction by obviously reducing the expression of phosphorylated tau and neuronal loss in vivo, which was deserved as a valuable molecular tool to reveal the role of DYRK1A in the pathogenesis of AD and to further promote the development of anti-AD drugs.
Asunto(s)
Enfermedad de Alzheimer , Quinasas DyrK , Inhibidores de Proteínas Quinasas , Proteínas Serina-Treonina Quinasas , Proteínas Tirosina Quinasas , Proteínas tau , Enfermedad de Alzheimer/tratamiento farmacológico , Enfermedad de Alzheimer/metabolismo , Humanos , Proteínas Tirosina Quinasas/antagonistas & inhibidores , Proteínas Tirosina Quinasas/metabolismo , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Proteínas Serina-Treonina Quinasas/metabolismo , Animales , Inhibidores de Proteínas Quinasas/farmacología , Inhibidores de Proteínas Quinasas/química , Inhibidores de Proteínas Quinasas/uso terapéutico , Inhibidores de Proteínas Quinasas/farmacocinética , Proteínas tau/metabolismo , Proteínas tau/antagonistas & inhibidores , Relación Estructura-Actividad , Administración Oral , Masculino , Ratas , Barrera Hematoencefálica/metabolismo , Barrera Hematoencefálica/efectos de los fármacos , Células HEK293 , Ratones , Descubrimiento de Drogas , Fosforilación/efectos de los fármacos , Simulación del Acoplamiento Molecular , Ratas Sprague-DawleyRESUMEN
Cumulus oophorus complexes (COCs) are the first extracellular barriers that sperm must pass through to fuse with oocytes, which have an important role in oocyte maturation and fertilization. However, little is known about the molecular mechanisms of COCs involved in fertilization. In this study, COCs were collected and then randomly divided into a test group that interacted with sperm and a control group that did not interact with sperm. Then, the total RNA was extracted; RNA transcriptome and small RNA libraries were prepared, sequenced, and analyzed. The results showed that 1283 differentially expressed genes (DEGs), including 560 upregulated and 723 downregulated genes. In addition, 57 differentially expressed miRNAs (DEMIs) with 35 upregulated and 22 downregulated were also detected. After the RNA-seq results were verified by RT-qPCR, 86 effective DEGs and 40 DEMIs were finally screened and a DEMI-DEG regulatory network was constructed. From this, the top ten hub target genes were HNF4A, SPN, WSCD1, TMEM239, SLC2A4, E2F2, SIAH3, ADORA3, PIK3R2, and GDNF, and they were all downregulated. The top ten hub DEMIs were miR-6876-5p, miR-877-3p, miR-6818-5p, miR-4690-3p, miR-6789-3p, miR-6837-5p, miR-6861-5p, miR-4421, miR-6501-5p, and miR-6875-3p, all of which were upregulated. The KEGG signaling pathway enrichment analysis showed that the effective DEGs were significantly enriched in the calcium, AMPK, and phospholipase D signaling pathways. Our study identified several DEGs and DEMIs and potential miRNA-mRNA regulatory pathways in COCs and these may contribute to fertilization. This study may provide novel insights into potential biomarkers for fertilization failure.
Asunto(s)
Células del Cúmulo , Redes Reguladoras de Genes , MicroARNs , ARN Mensajero , MicroARNs/genética , MicroARNs/metabolismo , Femenino , Animales , ARN Mensajero/metabolismo , ARN Mensajero/genética , Células del Cúmulo/metabolismo , Fertilización/genética , Masculino , Perfilación de la Expresión Génica , Transcriptoma , Ratones , Regulación de la Expresión GénicaRESUMEN
Multitarget-directed ligands (MTDLs) have recently attracted significant interest due to their superior effectiveness in multifactorial Alzheimer's disease (AD). Combined inhibition of two important AD targets, glycogen synthase kinase-3ß (GSK-3ß) and dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A), may be a breakthrough in the treatment of AD. Based on our previous work, we have designed and synthesized a series of novel harmine derivatives, investigated their inhibition of GSK-3ß and DYRK1A, and evaluated a variety of biological activities. The results of the experiments showed that most of these compounds exhibited good activity against GSK-3ß and DYRK1A in vitro. ZLQH-5 was selected as the best compound due to the most potent inhibitory effect against GSK-3ß and DYRK1A. Molecular docking studies demonstrated that ZLQH-5 could form stable interactions with the ATP binding pocket of GSK-3ß and DYRK1A. In addition, ZLQH-5 showed low cytotoxicity against SH-SY5Y and HL-7702, good blood-brain barrier permeability, and favorable pharmacokinetic properties. More importantly, ZLQH-5 also attenuated the tau hyperphosphorylation in the okadaic acid SH-SY5Y cell model. These results indicated that ZLQH-5 could be a promising dual-target drug candidate for the treatment of AD.
Asunto(s)
Enfermedad de Alzheimer , Neuroblastoma , Humanos , Enfermedad de Alzheimer/tratamiento farmacológico , Enfermedad de Alzheimer/metabolismo , Glucógeno Sintasa Quinasa 3 beta , Harmina/farmacología , Harmina/uso terapéutico , Proteínas tau/metabolismo , Proteínas tau/uso terapéutico , Simulación del Acoplamiento Molecular , Relación Estructura-Actividad , FosforilaciónRESUMEN
Cryopreservation of a small number of human spermatozoa is still a major challenge for embryologists. The aim of this study was to evaluate the clinical pregnancy and neonatal outcomes of intracytoplasmic sperm injection (ICSI) using a modified micro cryotube as freezing carrier for freezing small numbers of human spermatozoa collected by testicular sperm aspiration (TESA). We conducted a retrospective study to analyses the ICSI outcomes of using frozen-thawed few testicular spermatozoa in males with obstructive azoospermia (OA) from June 2017 to June 2021. Of 155 ICSI treatment cycles, 79 cycles were allocated to frozen sperm group and a modified micro cryotube was used for freezing testicular sperm, 76 cycles were allocated as fresh sperm group. No significant differences were observed in fertilization rate, good quality embryo rate, and blastocyst rate between the frozen sperm group and fresh sperm group (P > 0.05). Similarly, in the fresh embryo transfer cycles plus the first frozen-thawed embryo transfer cycles, the total clinical pregnancy rate (54.43% vs 57.89%), implantation rate (46.08% vs 49.47%), miscarriage rate (13.95% vs 13.64%) and live birth rate (45.57% vs 48.68%) were not statistically different between the frozen and fresh sperm groups (P > 0.05). In addition, there was no statistical differences in the mean gestational age (38.33weeks ± 1.74 vs 37.89weeks ± 1.87), preterm delivery rate (5.56% vs 10.81%), mean birth weight at delivery (3026.50 g ± 577.64 vs 2977.56 g ± 528.93), and low birth weight (12.50% vs 19.51%) between the two groups (P > 0.05 in all cases). Modified micro cryotube for cryopreservation of rare testicula rretrieved spermatozoa did not negatively affect the pregnancy and neonatal outcomes in TESA-ICSI cycles. The presented method may be a useful alternative for cryopreservation of small numbers of human spermatozoa in clinical setting.
Asunto(s)
Inyecciones de Esperma Intracitoplasmáticas , Recuperación de la Esperma , Embarazo , Femenino , Recién Nacido , Masculino , Humanos , Adulto , Inyecciones de Esperma Intracitoplasmáticas/métodos , Estudios Retrospectivos , Criopreservación/métodos , Semen , Espermatozoides , Índice de EmbarazoRESUMEN
Alzheimer's disease (AD) is characterized by progressive cognitive impairment and mental behavior. The combination inhibition of two essential AD targets, acetylcholinesterase (AChE) and glycogen synthase kinase-3ß (GSK-3ß), might be a breakthrough in the discovery of therapeutic success. Herein, 17 ß-carboline-1,2,3-triazole hybrids were designed, synthesized, and evaluated for their AChE and GSK-3ß inhibitory potential. The results indicated that compound 21 has the most potent inhibition against eeAChE (IC50 = 0.20 ± 0.02 µM), hAChE (IC50 = 0.34 ± 0.01 µM) and GSK-3ß (IC50 = 1.14 ± 0.05 µM) among these compounds. In addition, it inhibited hAChE in a mixed type manner and could occupy the binding pocket forming diverse interactions with the target of AChE and GSK-3ß. Moreover, compound 21 showed low cytotoxicity against SH-SY5Y and HepG2 cell lines and good BBB permeability. Compound 21 also attenuated the tau hyperphosphorylation in the Tau (P301L) 293T cell model. The ADME projection exhibited that compound 21 has acceptable physicochemical characteristics. This study provides new leads for the assessment of AChE and GSK-3ß dual inhibition as a promising strategy for AD treatment.
Asunto(s)
Enfermedad de Alzheimer , Neuroblastoma , Humanos , Enfermedad de Alzheimer/tratamiento farmacológico , Enfermedad de Alzheimer/metabolismo , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Inhibidores de la Colinesterasa/farmacología , Inhibidores de la Colinesterasa/uso terapéutico , Triazoles/farmacología , Triazoles/uso terapéutico , Acetilcolinesterasa/metabolismo , Carbolinas/farmacología , Carbolinas/uso terapéutico , Proteínas tau/metabolismo , FosforilaciónRESUMEN
Multi-targeted directed ligands (MTDLs) are emerging as promising Alzheimer's disease (AD) therapeutic possibilities. Coumarin is a multifunctional backbone with extensive bioactivity that has been utilized to develop innovative anti-neurodegenerative properties and is a desirable starting point for the construction of MTDLs. Herein, we explored and synthesized a series of novel coumarin derivatives and assessed their inhibitory effects on cholinesterase (AChE, BuChE), GSK-3ß, and BACE1. Among these compounds, compound 30 displayed the multifunctional profile of targeting the AChE (IC50 = 1.313 ± 0.099 µM) with a good selectivity over BuChE (SI = 24.623), GSK-3ß (19.30% inhibition at 20 µM), BACE1 (IC50 = 1.227 ± 0.112 µM), along with moderate HepG2 cytotoxicity, SH-SY5Y cytotoxicity, low HL-7702 cytotoxicity, as well as good blood-brain barrier (BBB) permeability. Kinetic and docking studies indicated that compound 30 was a competitive AChE inhibitor. Furthermore, acute toxicity experiments revealed that it was non-toxic at a dosage of 1000 mg/kg. The ADME prediction results indicate that 30 has acceptable physicochemical properties. Collectively, these findings demonstrated that compound 30 would be a potential multifunctional candidate for AD therapy.
Asunto(s)
Enfermedad de Alzheimer , Neuroblastoma , Acetilcolinesterasa/metabolismo , Enfermedad de Alzheimer/tratamiento farmacológico , Secretasas de la Proteína Precursora del Amiloide , Ácido Aspártico Endopeptidasas , Inhibidores de la Colinesterasa/química , Colinesterasas/metabolismo , Cumarinas/química , Diseño de Fármacos , Glucógeno Sintasa Quinasa 3 beta , Humanos , Ligandos , Neuroblastoma/tratamiento farmacológico , Relación Estructura-ActividadRESUMEN
Two new pyranone derivatives phomapyrone A (2) and phomapyrone B (3), one new coumarin 11S, 13R-(+)-phomacumarin A (1), three known pyranones (4-6), together with three known amide alkaloids fuscoatramides A-C (7-9), as well as 9S, 11R-(+)-ascosalitoxin (10) were isolated from the endophytic fungus Phoma sp. YN02-P-3, which was isolated from the healthy leaf tissue of a Paulownia tree in Yunnan Province, China. Their structures were elucidated using extensive NMR spectroscopic and HRESIMS data and by comparing the information with literature data. In addition, all compounds were tested for their cytotoxicity activity against human tumor cell lines, and the results showed that new compounds 1-3 showed moderate inhibitory activity against the HL-60 cell line with IC50 values of 31.02, 34.62, and 27.90 µM, respectively.
RESUMEN
Alzheimer's disease (AD) is a chronic and progressive neurodegenerative disease, characterized by irreversible cognitive impairment, memory loss, and behavioral disturbances, ultimately resulting in death. The critical roles of glycogen synthase kinase-3ß (GSK-3ß) in tau pathology have also received considerable attention. Based on molecular docking studies, a series of novel α-carboline derivatives were designed, synthesized, and evaluated as GSK-3ß inhibitors for their various biological activities. Among them, compound ZCH-9 showed the most potent inhibitory activity against GSK-3ß, with an IC50 value of 1.71 ± 0.09 µM. The cytotoxicity assay showed that ZCH-9 had low cytotoxicity toward the cell lines SH-SY5Y, HepG2, and HL-7702. Moreover, Western blot analysis indicated that ZCH-9 effectively inhibited hyperphosphorylation of the tau protein in okadaic acid-treated SH-SY5Y cells. The binding mode between ZCH-9 and GSK-3ß was analyzed and further clarified throughout the molecular dynamics simulations. In general, these results suggested that the α-carboline-based small-molecule compounds could serve as potential candidates targeting GSK-3ß for the treatment of AD.
Asunto(s)
Enfermedad de Alzheimer , Neuroblastoma , Enfermedades Neurodegenerativas , Enfermedad de Alzheimer/tratamiento farmacológico , Enfermedad de Alzheimer/metabolismo , Carbolinas , Glucógeno Sintasa Quinasa 3 beta , Humanos , Simulación del Acoplamiento Molecular , Ácido Ocadaico/metabolismo , Fosforilación , Relación Estructura-Actividad , Proteínas tau/metabolismoRESUMEN
Epigenetics regulate the gene expression and chromatin organization associated with the development and occurrence of cancer. Histone deacetylase inhibitors (HDACis) have been proved to be an effective epigenetic targeting drug for cancer treatment. The structures of most HDACis were divided into four parts, including cap group, connection unit, linker region and zinc binding group. We designed a series of compounds containing the structure of phenoxyacetate for the linker region and cyclohexene for connection unit as a novel type of inhibitors. Representative compound YZ1 exhibited obvious antiproliferative activity against four different cell lines and potent enzymatic inhibitory activities to class I HDACs, which IC50 of HDAC1-3 were 1.6 nM, 1.9 nM and 3.8 nM respectively. In addition, YZ1 concentration-dependently inhibited cell proliferation, induced apoptosis and cycle arrest at G2/M phase in HCT116 cells. With biological activity assessment and docking studies, these results indicate YZ1 has the potential to be a lead compound for further optimization as HDAC inhibitors.
Asunto(s)
Antineoplásicos , Inhibidores de Histona Desacetilasas , Aminas/farmacología , Antineoplásicos/química , Línea Celular Tumoral , Proliferación Celular , Diseño de Fármacos , Ensayos de Selección de Medicamentos Antitumorales , Inhibidores de Histona Desacetilasas/química , Humanos , Simulación del Acoplamiento Molecular , Relación Estructura-ActividadRESUMEN
Uncontrolled cell proliferation is a hallmark of cancer. The major regulator of the cell cycle, cyclin dependent kinase 2 (CDK2), has become a mature target for cancer treatment. Herein, we describe our efforts toward the discovery of a series of benzofuro[3,2-b]quinoline alkaloid derivatives as CDK2 inhibitors through a scaffold hopping strategy. Compound ZLHQ-5f has topoisomerase I (Topo I) inhibitory activity due to the unique structure of benzofurano[3,2-b]quinoline. Resultantly, ZLHQ-5f exhibited promising anti-proliferative and CDK2 inhibitory activities. It also arrests the cell cycle in S-phase, triggers apoptosis in HCT116 cells, and has a good safety profile in vivo. There has yet to be a report on dual CDK2/Topo I inhibitor, thus this will be a novel attempt.
Asunto(s)
Antineoplásicos , Quinolinas , Antineoplásicos/química , Línea Celular Tumoral , Proliferación Celular , Quinasa 2 Dependiente de la Ciclina , Células HCT116 , Humanos , Estructura Molecular , Quinolinas/química , Quinolinas/farmacología , Relación Estructura-Actividad , Inhibidores de Topoisomerasa I/química , Inhibidores de Topoisomerasa I/farmacologíaRESUMEN
The natural product harmine, a representative ß-carboline alkaloid from the seeds of Peganum harmala L. (Zygophyllaceae), possesses a broad spectrum of biological activities. In this study, a novel series of harmine derivatives containing N-benzylpiperidine moiety were identified for the treatment of Alzheimer's disease (AD). The results showed that all the derivatives possessed significant anti-acetylcholinesterase (AChE) activity and good selectivity over butyrylcholinesterase (BChE). In particular, compound ZLWH-23 exhibited potent anti-AChE activity (IC50 = 0.27 µM) and selective BChE inhibition (IC50 = 20.82 µM), as well as acceptable glycogen synthase kinase-3 (GSK-3ß) inhibition (IC50 = 6.78 µM). Molecular docking studies and molecular dynamics simulations indicated that ZLWH-23 could form stable interaction with AChE and GSK-3ß. Gratifyingly, ZLWH-23 exhibited good selectivity for GSK-3ß over multi-kinases and very low cytotoxicity towards SH-SY5Y, HEK-293T, HL-7702, and HepG2 cell lines. Importantly, ZLWH-23 displayed efficient reduction against tau hyperphosphorylation on Ser-396 site in Tau (P301L) 293T cell model. Collectively, harmine-based derivatives could be considered as possible drug leads for the development of AD therapies.
Asunto(s)
Acetilcolinesterasa/metabolismo , Enfermedad de Alzheimer/tratamiento farmacológico , Antineoplásicos/química , Carbolinas/síntesis química , Inhibidores de la Colinesterasa/síntesis química , Glucógeno Sintasa Quinasa 3 beta/antagonistas & inhibidores , Inhibidores de Proteínas Quinasas/síntesis química , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Carbolinas/farmacología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Inhibidores de la Colinesterasa/farmacología , Diseño de Fármacos , Humanos , Simulación del Acoplamiento Molecular , Simulación de Dinámica Molecular , Unión Proteica , Inhibidores de Proteínas Quinasas/farmacología , Transducción de Señal , Relación Estructura-ActividadRESUMEN
OBJECTIVES: Ultrasound-guided saline enema is highly successful in treating pediatric intussusception; however, early recurrence-within 48 hours-is possible. This study aimed to explore effective methods of reducing early recurrence. METHODS: This study included patients aged 0 to 14 years diagnosed with ileocolic intussusception with a symptom duration of <48 hours from January 2019 to March 2021. The patients were divided into control and intervention groups. All patients received successful treatment with ultrasound-guided saline enema; however, in patients treated before January 4, 2020 (control group), the intestinal fluid was drained immediately, and in patients treated after January 4, 2020 (intervention group), the intestinal fluid was drained after 15 minutes of intestinal pressure maintenance. Early recurrence rates of the groups were compared. RESULTS: Ileocolic intussusception was treated successfully by ultrasound-guided saline enema in 231 patients (116, control group;115, intervention group). The early recurrence rate in the intervention group (10%; 95% CI: 4.9-16.5) was numerically lower than that in the control group (19%; 95% CI: 12.3-27.3). No significant difference was observed in the number of recurrences per person between the groups (P = .448). Patients without early recurrence were older (P = .004) and received enemas of a shorter duration (P < .001) and lower pressure (P < .001) than patients without early recurrence. CONCLUSIONS: Maintaining reduction pressure for 15 minutes after a successful ultrasound-guided saline enema may reduce the early recurrence of intussusception. A randomized controlled trial is needed because the intervention and control cohorts were most probably incomparable (due to the COVID-19 pandemic).
Asunto(s)
COVID-19 , Enfermedades del Íleon , Intususcepción , Niño , Enema/métodos , Humanos , Enfermedades del Íleon/diagnóstico por imagen , Enfermedades del Íleon/terapia , Lactante , Intususcepción/diagnóstico por imagen , Intususcepción/terapia , Pandemias , Recurrencia , Estudios Retrospectivos , Solución Salina , Resultado del Tratamiento , Ultrasonografía IntervencionalRESUMEN
BACKGROUND: Azoospermic patients have benefited from both epididymal and testicular spermatozoa intracytoplasmic sperm injection (ICSI) treatment and lasers have been used to identify viable, immotile spermatozoa before the procedure. There are limited studies on the safety of laser-assisted selection of immotile spermatozoa. The aim of this study was to investigate the impact of laser-assisted selection of immotile spermatozoa on the obstetric and neonatal outcomes after ICSI. METHODS: A retrospective comparative study was conducted on outcomes of ICSI cycles with testicular spermatozoa from June 2014 to June 2018. Of 132 cycles, 33 were allocated to the test group and oocytes were injected with immotile spermatozoa selected by laser, 99 cycles were allocated as control group. RESULTS: Compared with the control group, no significant differences were found in the pregnancy, implantation, miscarriage and live birth rates in the test group in either fresh or frozen transfer cycles. The cumulative live birth rate in the test group was 69.70%, which was slightly higher than in the control group (60.61%), but this was not statistically different. There were no differences in the average gestational age, premature birth rate, neonatal birth weight, and the malformation rate between the test and control groups (P > 0.05). In addition, the obstetric outcome between the two groups were not different (P > 0.05). CONCLUSIONS: No negative effect on perinatal and neonatal outcomes was seen by using laser-assisted selection of immotile spermatozoa for TESA-ICSI. This study endorses the use of laser-assisted selection of viable spermatozoa for ICSI cycles.
Asunto(s)
Azoospermia/terapia , Separación Celular/métodos , Resultado del Embarazo , Inyecciones de Esperma Intracitoplasmáticas , Recuperación de la Esperma , Adulto , Azoospermia/epidemiología , Azoospermia/patología , Estudios de Casos y Controles , China/epidemiología , Femenino , Fertilización In Vitro/métodos , Humanos , Recién Nacido , Rayos Láser , Masculino , Embarazo , Resultado del Embarazo/epidemiología , Índice de Embarazo , Estudios Retrospectivos , Inyecciones de Esperma Intracitoplasmáticas/métodos , Motilidad EspermáticaRESUMEN
AIM: Although mammalian embryos could be preserved in liquid nitrogen for thousands of years in theoretical models, the viability of cryopreserved blastocyst with varying grades remains to be speculated. In this study, we aimed to determine whether the longer storage time of blastocysts with equal grades could negatively affect the perinatal outcomes. MATERIALS AND METHODS: Single vitrified-warmed blastocyst was divided into four grades (AA, AB/BA, BB, BC/CB) according to the blastocyst score when freezing, and each grade of blastocyst was categorized into four storage duration categories: 28 days-1 year, 1-3 years, 3-5 years, and ≥5 years. Then the perinatal outcomes with different storage time were analyzed. RESULTS: Our results revealed that for blastocysts with the same grade, the length of storage time had no statistical effect on blastocyst survival rate, clinical pregnancy/implantation rate, live birth rate, and abortion rate. In addition, more advanced developmental blastocyst could obtain better pregnancy outcomes regardless of the cryopreservation length. Similar neonatal outcomes were obtained over time. CONCLUSIONS: Cryopreservation time could not negatively affect the perinatal outcomes of blastocysts with equal grades. Efficient blastocyst cryopreservation technology by vitrification can help older women obtain high-quality embryos at a young age.
Asunto(s)
Criopreservación , Técnicas de Cultivo de Embriones , Anciano , Blastocisto , Criopreservación/métodos , Femenino , Humanos , Recién Nacido , Embarazo , Índice de Embarazo , Estudios Retrospectivos , VitrificaciónRESUMEN
Alzheimer's disease (AD) is a chronic and progressive neurodegenerative disease, characterized by irreversible cognitive impairment, memory loss and behavioral disturbances, ultimately leading to death. Glycogen synthase kinase 3ß (GSK-3ß) and dual-specificity tyrosine phosphorylation regulated kinase1A (DYRK1A) have gained a lot of attention for its role in tau pathology. To search for potential dual GSK-3ß/DYRK1A inhibitors, we focused on harmine, a natural ß-carboline alkaloid, which has been extensively studied for its various biological effects on the prevention of AD. In this study, a new series of harmine derivatives were designed, synthesized and evaluated as dual GSK-3ß/DYRK1A inhibitors for their multiple biological activities. The in vitro results indicated that most of them displayed promising activity against GSK-3ß and DYRK1A. Among them, compound ZDWX-25 showed potent inhibitory effects on GSK-3ß and DYRK1A with IC50 values of 71 and 103 nM, respectively. Molecular modelling and kinetic studies verified that ZDWX-25 could interact with the ATP binding pocket of GSK-3ß and DYRK1A. Western blot analysis revealed that ZDWX-25 inhibited hyperphosphorylation of tau protein in okadaic acid (OKA)-induced SH-SY5Y cells. In addition, ZDWX-25 showed good blood-brain barrier penetrability in vitro. More importantly, ZDWX-25 could ameliorate the impaired learning and memory in APP/PS1/Tau transgenic mice. These results indicated that the harmine-based compounds could be served as promising dual-targeted candidates for AD.
Asunto(s)
Enfermedad de Alzheimer/tratamiento farmacológico , Glucógeno Sintasa Quinasa 3 beta/antagonistas & inhibidores , Harmina/farmacología , Fármacos Neuroprotectores/farmacología , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Proteínas Tirosina Quinasas/antagonistas & inhibidores , Enfermedad de Alzheimer/metabolismo , Línea Celular , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Diseño de Fármacos , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Harmina/síntesis química , Harmina/química , Humanos , Modelos Moleculares , Estructura Molecular , Fármacos Neuroprotectores/síntesis química , Fármacos Neuroprotectores/química , Inhibidores de Proteínas Quinasas/síntesis química , Inhibidores de Proteínas Quinasas/química , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Tirosina Quinasas/metabolismo , Relación Estructura-Actividad , Quinasas DyrKRESUMEN
The aim of this study was to provide guidance for better management in the selection of blastocyst to warm in frozen-thawed embryo transfer (FET) cycles. A retrospective cohort follow-up study was conducted that included single autologous frozen blastocyst transfer cycles performed in our Reproductive Medicine Unit from January 2009 to December 2016. The live birth rate (LBR), clinical pregnancy rate (cPR) were increased as blastocyst morphology scores increased, but the miscarriage rate decreased in all groups. In the high-score groups, there were no differences in LBR between D5 and D6, while in the low-score groups, LBR was significantly higher in D5 compared to the D6. With respect to neonatal outcome, there were no differences in all the groups. After binary logistic regression analysis, it was seen that patients' age, thawed cycles, pre-frozen morphology score and developmental rate were independently associated with LBR. These results suggest that for high-scoring blastocyst, the pre-frozen morphological score should be given priority while for low-scoring blastocysts, the developmental rate should be given priority when thawing in FET cycles.
RESUMEN
A viable spermatozoon is a prerequisite for fertilization in intracytoplasmic sperm injection (ICSI). Thus, it is crucial to select viable but immotile spermatozoa on the day of ICSI. We report conflicting results in the identification of viable but immotile spermatozoa between the eosin-nigrosin staining and the laser test, which resulted in confusion for embryologists during assisted reproductive technology (ART). Three patients' semen samples that showed no motile spermatozoa are described in this report. To identify viable spermatozoa, we used both the eosin-nigrosin test and the laser test for each sample, and repeated the semen analysis twice in each patient. Viable but immotile spermatozoa selected by the laser test were used for ICSI. Viable spermatozoa were detected by both the eosin-nigrosin and laser tests in two patients (case 1, 95.00% vs. 24.21% and 92.68% vs. 22.22%; case 2, 41.18% vs. 23.48% and 39.81% vs. 22.52%), indicating consistent results between the two methods. In the third patient, the eosin-nigrosin test yielded viability rates of 20.75% and 19.14%, while the result of the laser test was 0%. Thus, testicular aspiration was performed to collect viable sperm from this patient. Normal fertilization was achieved after the injection of viable but immotile spermatozoa selected from these patients by the laser test, resulting in the birth of two healthy babies. Our study documents a case where the eosin-nigrosin test showed a limitation in identifying viable but immotile spermatozoa for ART, while the laser test may overcome this limitation. Larger samples may be required to corroborate the clinical value of the laser test.
RESUMEN
Intracytoplasmic sperm injection (ICSI) remains the most effective method for severe male infertility patients to obtain their genetic offspring. A viable spermatozoon is the prerequisite for initiating fertilization in ICSI. Motility is the primary sign of sperm viability. However, how to select immotile but viable spermatozoa for ICSI on the day of ovum pick-up is critical for ICSI. This review focuses on the techniques for the identification and selection of immotile but viable spermatozoa for assisted reproductive technology.
Asunto(s)
Inyecciones de Esperma Intracitoplasmáticas , Recuperación de la Esperma , Espermatozoides , Humanos , Infertilidad Masculina , Masculino , Motilidad EspermáticaRESUMEN
OBJECTIVE: To investigate the effectiveness of cumulus oophorus complexes (COCs) in the physiologic selection of spermatozoa for intracytoplasmic sperm injection (ICSI). DESIGN: A prospective sibling oocytes study. SETTING: Center of reproductive medicine. PATIENT(S): Couples undergoing ICSI during 2016, females aged ≤38 years, and at least six metaphase II (MII) oocytes retrieved. Sixty patients were included in the study. Of 857 MII oocytes, 429 were allocated to the study group and were injected with the sperm selected via COCs; 428 MII oocytes were allocated as controls (C) and fertilized by conventional ICSI. INTERVENTION(S): In the study group, ICSI was performed with spermatozoa that traversed the COCs in vitro. MAIN OUTCOMES MEASURE(S): Blastocyst/top blastocyst formation rate, fertilization rate, and oocyte utilization rate. RESULT(S): Oocytes injected with COC-selected spermatozoa had a significantly higher fertilization rate than the conventional ICSI group (85.31% vs. 74.77%). There were no statistically differences in cleavage and top embryo rate on day 3 between the COC-ICSI and C-ICSI groups. However, with day 5 or 6 embryos, compared with conventional ICSI, COC-ICSI significantly improved blastocyst formation rate (64.90% vs. 53.50%), blastocyst formation rate at day 5 (46.52% vs. 38.85%), top blastocyst rate (38.72% vs. 24.20%), and the usable blastocysts formation rate (62.12% vs. 46.82%). The oocyte utilization rate was improved greatly in the COC-ICSI group compared with the C-ICSI group (51.98% vs. 34.35%). Furthermore, the fertilization rate, top embryo rate on day 3, usable blastocyst rate, top blastocyst rate, and day 5 usable blastocysts rate were similar between the conventional IVF and COC-ICSI groups. Single-blastocyst transfer was performed in 82 cycles, including 44 fresh cycles and 38 frozen-thawed cycles. The cumulative embryo implantation rate in the COC-ICSI group was 64.29%, slightly higher than in the C-ICSI group (53.85%), but there was no statistical difference. CONCLUSION(S): The use of COCs to select spermatozoa for ICSI appears to be effective and led to a statistically significant improvement in blastocyst development and quality.
Asunto(s)
Células del Cúmulo/fisiología , Infertilidad Femenina/terapia , Inyecciones de Esperma Intracitoplasmáticas/métodos , Espermatozoides/fisiología , Adulto , Transferencia de Embrión/métodos , Transferencia de Embrión/tendencias , Femenino , Humanos , Infertilidad Femenina/diagnóstico , Masculino , Oocitos/fisiología , Estudios Prospectivos , Distribución AleatoriaRESUMEN
BACKGROUND: Sperm cryopreservation is the most effective method to preserve male fertility but this is normally used for motile spermatozoa. Thus, only motile spermatozoa are used for cryopreservation in most reproductive medicine centers worldwide. The immotile spermatozoa from some problematic patients are usually discarded, resulting in a missed opportunity of sterility cryopreservation for future assisted reproductive treatments. Many studies have shown that successful fertilization can be obtained after selection of viable sperm from the completely immotile spermatozoa before ICSI. Whether the completely immotile spermatozoa are worth of freezing has not been realized The aim of this study is to explore the clinical value of cryopreservation of immotile spermatozoa. METHODS: Completely immotile spermatozoa were collected and frozen, and subsequently viable but immotile frozen-thawed spermatozoa were selected by laser plus for ICSI. Main outcomes included spermatozoa survival index, fertilization rate and good quality embryo rate. RESULTS: After identification by laser, the fresh samples of spermatozoa presented with a mean survival rate of 54.86% and 26.05%, and this was reduced to 44.13% and 18.13% in frozen-thawed spermatozoa samples, which showed a frozen-thawed spermatozoa survival index of 0.80 and 0.70 in the testicular and ejaculate sperm, respectively. There were no statistically differences in fertilization rate (80% vs80.51%, 75.00% vs 81.48%), cleavage rate (95.45% vs 98.95%, 100.00% vs 95.45%) and good quality embryo rate (40.48% vs 52.13%, 33.33%vs38.10%) between the frozen-thawed immotile spermatozoa group and the routine fresh immotile spermatozoa ICSI group in both testicular and ejaculate sperm, respectively. CONCLUSIONS: The results of the study show that completely immotile spermatozoa can be frozen in order to preserve male fertility as long as viable spermatozoa are present. This procedure provides a further possibility for fertility preservation for patients with completely immotile spermatozoa.