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The complexity of microvascular circulation has led to the development of advanced imaging techniques and biomimetic models. This study developed a multifaceted microfluidic-based microdevice as an in vitro model of microvasculature to replicate important geometric and functional features of in vivo perfusion in mice. The microfluidic device consisted of a microchannel for blood perfusion, mirroring the natural hierarchical branching vascular structures found in mice. Additionally, the device incorporated a steady gradient of oxygen (O2) which diffused through the polydimethylsiloxane (PDMS) layer, allowing for dynamic blood oxygenation. The assembled multi-layered microdevice was accompanied by a dual-modal imaging system that combined laser speckle contrast imaging (LSCI) and intrinsic signal optical imaging (ISOI) to visualize full-field blood flow distributions and blood O2 profiles. By closely reproducing in vivo blood perfusion and oxygenation conditions, this microvasculature model, in conjunction with numerical simulation results, can provide quantitative information on physiologically relevant hemodynamics and key O2 transport parameters that are not directly measurable in traditional animal studies.
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Hemodinámica , Microfluídica , Ratones , Animales , Oxígeno , MicrovasosRESUMEN
BACKGROUND: Alcohol use disorders result from repeated binge and chronic alcohol consumption followed by negative effects, such as anxiety, upon cessation. This process is associated with the activation of NLRP3 inflammasome-mediated responses. However, whether and how inhibition of the NLRP3 inflammasome alters alcohol intake and anxiety behavior remains unclear. METHODS: A combination of drinking-in-the-dark and gavage was established in NLRP3-knockout and control mice. Behavior was assessed by open-field and elevated plus maze tests. Binge alcohol drinking was measured at 2 h and 4 h. A 2 h/4 h/24 h voluntary drinking was determined by a two-bottle choice paradigm. Western blotting and ELISA were applied to examine the levels of the NLRP3 inflammasome and- inflammatory factors, such as IL-1ß and TNF-α. Nissl staining was used to measure neuronal injury. The electrophysiological method was used to determine glutamatergic transmission in corticostriatal circuits. In vivo optogenetic LTP and LTD were applied to control the function of corticostriatal circuits on the behavior of mice. MCC950 was used to antagonize the NLRP3 inflammasome. RESULTS: The binge alcohol intake was decreased in NLRP3 KO mice compared to the control mice. During alcohol withdrawal, NLRP3 deficiency attenuated anxiety-like behavior and neuronal injury in the mPFC and striatum. Moreover, we discovered that glutamatergic transmission to striatal neurons was reduced in NLRP3 KO mice. Importantly, in vivo optogenetic induction of long-term potentiation (LTP) of corticostriatal circuits reversed the effects of NLRP3 deficiency on glutamatergic transmission and anxiety behavior. We also demonstrated that optogenetic induction of LTD decreased anxiety-like behavior and caused a reduction in glutamatergic transmission. Interestingly, NLRP3 deficiency or inhibition (MCC950 injection) attenuated the anxiety-like behavior, but it did not prevent DID + gavage paradigm-induced a persistent enhancement of drinking in a two-bottle choice at 2 and 4 days into withdrawal. CONCLUSION: Our results demonstrate that NLRP3 deficiency decreases binge alcohol intake and anxiety-like behavior through downregulation of glutamatergic transmission in corticostriatal circuits, which may provide an anti-inflammatory target for treating alcohol use disorders.
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Alcoholismo , Síndrome de Abstinencia a Sustancias , Ratones , Animales , Inflamasomas , Proteína con Dominio Pirina 3 de la Familia NLR , Ansiedad , Consumo de Bebidas Alcohólicas , Ratones Endogámicos C57BLRESUMEN
Long-term opioid abuse causes a variety of long-lasting cognitive impairments such as attention, impulsivity and working memory. These cognitive impairments undermine behavioural treatment for drug abuse and lead to poor treatment retention and outcomes. Modafinil is a wake-promoting drug that shows potential in improving attention and memory in humans and animals. However, modafinil's effect on opioid-induced cognitive impairments remains unclear, and the underlying mechanism is poorly understood. This study showed that repeated morphine administration significantly impairs attention, increases impulsivity and reduces motivation to natural rewards in mice. Systemic modafinil treatment at low dose efficiently ameliorates morphine-induced attention dysfunction and improves motivation and working memory in mice. High dose of modafinil has adverse effects on impulsive action and attention. Local infusion of D1R antagonist SCH-23390 reverses the morphine-induced synaptic abnormalities and activation of the D1R-ERK-CREB pathway in medial prefrontal cortex (mPFC). This study demonstrated a protective effect of modafinil in mPFC neurons and offered a therapeutic potential for cognitive deficits in opioid abuse.
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Atención/efectos de los fármacos , Trastornos del Conocimiento/fisiopatología , Modafinilo/farmacología , Morfina/farmacología , Corteza Prefrontal/efectos de los fármacos , Animales , Trastornos del Conocimiento/inducido químicamente , Relación Dosis-Respuesta a Droga , Conducta Impulsiva/efectos de los fármacos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Ratones , Modafinilo/administración & dosificación , Modafinilo/efectos adversos , Motivación/efectos de los fármacosRESUMEN
Alzheimer's disease (AD) is a chronic central neurodegenerative disease. The pathological features of AD are the extracellular deposition of senile plaques formed by amyloid-ß oligomers (AßOs) and the intracellular accumulation of neurofibrillary tangles formed by hyperphosphorylated tau protein. In this paper, an in vitro pathological model of AD based on neuronal network chip and its real-time dynamic analysis were presented. The hippocampal neuronal network was cultured on the microelectrode array (MEA) chip and induced by AßOs as an AD model in vitro to simultaneously record two firing patterns from the interneurons and pyramidal neurons. The spatial firing patterns mapping and cross-correlation between channels were performed to validate the degeneration of neuronal network connectivity. This biosensor enabled the detection of the AßOs toxicity responses, and the identification of connectivity and interactions between neuronal networks, which can be a novel technique in the research of AD pathological model in vitro.
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Enfermedad de Alzheimer , Péptidos beta-Amiloides , Humanos , Ovillos Neurofibrilares , Proteínas tauRESUMEN
It has previously been demonstrated that ischemic stroke activates autophagy pathways; however, the mechanism remains unclear. The aim of this study is to further investigate the role that autophagy plays in cerebral ischemia. 2, 4-diamino-6-hydroxy-pyrimidine (DAHP), for its nitric oxide synthase (NOS) inhibiting neuroprotective effect, and triptolide (TP), for its anti-inflammatory property, were selected to administer pre middle cerebral artery occlusion (MCAO). The drugs were administered 12 hours prior to MCAO. Both magnetic resonance imaging (MRI) and 2, 3, 5-triphenyltetrazolium chloride (TTC) staining showed that the drugs reduce the area of infarction. Immunoblotting analysis revealed increases in Beclin-1 and myeloid cell leukelia-1(Mcl-1) in treated rats. This could be a contributing factor to the reduction in autophagy induced damage. Immunochemistry and western blot showed that mTOR expression in treated rats was marginally different 24 h after injury, and this could also be significant in the mechanism. Furthermore, terminal deoxynucleotidyl transferase- (TdT-) mediated dUTP nick end labeling (TUNEL) staining proved that the drugs are effective in reducing apoptosis. The upregulation of Beclin-1 and Mcl-1 and downregulation of Bcl-2, caspase-3, and the Bcl-2/Beclin-1 ratio infer that the neuroprotective effect of DAHP and TP act via the mediation of autophagy and apoptosis pathways.
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Autofagia/fisiología , Isquemia Encefálica/tratamiento farmacológico , Diterpenos/uso terapéutico , Fenantrenos/uso terapéutico , Azúcares Ácidos/uso terapéutico , Animales , Apoptosis/efectos de los fármacos , Autofagia/genética , Western Blotting , Compuestos Epoxi/uso terapéutico , Inmunohistoquímica , Etiquetado Corte-Fin in Situ , Espectroscopía de Resonancia Magnética , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
Understanding neural pathways and cognitive processes involved in the transformation of dietary fats into sensory experiences has profound implications for nutritional well-being. This study presents an efficient approach to comprehending the neural perception of fat taste using electroencephalogram (EEG). Through the examination of neural responses to different types of fatty acids (FAs) in 45 participants, we discerned distinct neural activation patterns associated with saturated versus unsaturated fatty acids. The spectrum analysis of averaged EEG signals revealed notable variations in δ and α-frequency bands across FA types. The topographical distribution and source localization results suggested that the brain encodes fat taste with specific activation timings in primary and secondary gustatory cortices. Saturated FAs elicited higher activation in cortical associated with emotion and reward processing. This electrophysiological evidence enhances our understanding of fundamental mechanisms behind fat perception, which is helpful for guiding strategies to manage hedonic eating and promote balanced fat consumption.
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Encéfalo , Grasas de la Dieta , Electroencefalografía , Percepción del Gusto , Humanos , Femenino , Adulto Joven , Adulto , Masculino , Encéfalo/fisiología , Grasas de la Dieta/metabolismo , Grasas de la Dieta/análisis , Gusto , Ácidos Grasos/química , Ácidos Grasos/metabolismoRESUMEN
Mammalian olfactory perception is an important physiological function for tasks such as finding food, identifying species, and avoiding enemies. Previous studies have demonstrated the molecular basis of receptors and the anatomical structure of the olfactory bulb (OB), which is the initial processing center for odor perception. However, there remains some controversy about the coding and transmission mode of olfactory information in the OB through neuronal interaction mediated by different neurotransmitters. In this paper, a biomimetic sensor based on OB neuronal network was developed to detect trace amounts of typical neurotransmitters and decode odor perception in vitro. Primary OB neurons were seeded on a microelectrode array (MEA) chip to capture multichannel extracellular electrical activities. The firing features of OB neurons were statistically analyzed after stimulation with graded concentrations of the glutamate and gamma-aminobutyric acid (GABA). Cross-correlation analysis between channels was used to evaluate the connection status in the neuronal networks. The concentration-dependent responses to these two neurotransmitters were assessed over a range of values, and the lower limits of detection for glutamate and GABA were 100 nM and 50 nM, respectively. Stimulation with excessive concentrations produced response tolerance and neurotoxicity, and transmission between cells in the neural network was modulated by different neurotransmitters. This device could serve as a novel biosensor for detecting trace amounts of common neurotransmitters and for screening the effects of drugs on the physiology of olfaction. The response patterns of this biomimetic sensor are conducive to revealing the coding mechanism of information processing in the olfactory system.
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Técnicas Biosensibles , Percepción Olfatoria , Animales , Biomimética , Ácido Glutámico , Mamíferos , Neurotransmisores , Odorantes/análisis , Bulbo Olfatorio/fisiología , Percepción Olfatoria/fisiología , Olfato , Ácido gamma-AminobutíricoRESUMEN
Olfactory dysfunction is an early symptom of neurodegenerative disease. Amyloid-beta oligomers (AßOs), the pathologic protein of Alzheimer's disease (AD), have been confirmed to be firstly deposited in olfactory bulb (OB), causing smell to malfunction. However, the detailed mechanisms underlying pathogenic nature of AßOs-induced olfactory neuronal degeneration in AD are not completely realized. Here, an early-stage olfactory dysfunction pathological model of AD in vitro based on biomimetic OB neuronal network chip was established for dynamic multi-site detection of neuronal electrical activity and network connection. We found both spike firing and correlation of overall neuronal network change regularly displayed gradually active state and then rapidly decay state after AßOs induction. Moreover, MK-801 and memantine were administrated at early-stage to detect alteration of OB neurons simulating nasal administration for AD treatment, which showed an almost recovery through the intermittent firing pattern. Together, this neuronal network-on-chip has revealed synaptic impairment and network neurodegeneration of olfactory dysfunction in AD, providing potential mechanisms information for early-stage progressive olfactory amyloidogenic pathology.
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Enfermedad de Alzheimer , Técnicas Biosensibles , Enfermedades Neurodegenerativas , Trastornos del Olfato , Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/metabolismo , Biomimética , Maleato de Dizocilpina/metabolismo , Humanos , Memantina/metabolismo , Neuronas/metabolismo , Trastornos del Olfato/etiología , Trastornos del Olfato/metabolismo , Trastornos del Olfato/patología , Bulbo Olfatorio , OlfatoRESUMEN
RATIONALE: Opioid use disorder is a complicated brain disease with high heritability. The underlying mechanisms of the genetic underpinnings in the susceptibility and treatment response of opioid use disorder remain elusive. OBJECTIVES: To reveal the potential associations of genotypes and gene methylations of dopaminergic system genes, as well as roles of them in opioid use disorder. In the present study, we detected the DNA methylation in the promoter regions of five representative dopaminergic system genes (DRD1, DRD2, SLC6A3, TH, and COMT) between 120 patients with heroin use disorder in methadone maintenance treatment (MMT) program and 111 healthy controls. The associations of 25 SNPs in the above genes and methylation of 237 CpG sites, known as methylation quantitative trait loci (mQTLs), were determined. Then, the correlations of the above mQTLs and traits of heroin use disorder were analyzed in a sample set of 801 patients with heroin use disorder and 930 healthy controls. RESULTS: Our results demonstrated that several mQTLs in the DRD1 and DRD2 genes were identified both in the heroin use disorder and healthy control groups. Interestingly, rs4867798-CpG_174872884 and rs5326-CpG_174872884 in the DRD1 gene were the unique SNP-CpG pairs in the patients with heroin use disorder. Furthermore, mQTL rs5326 was associated with the susceptibility and effective dosage of MMT for heroin use disorder, and demonstrated allele-specific correlation with the expression of the DRD1 gene in the human caudate. CONCLUSIONS: Our findings suggest that some mQTLs may be associated with traits of opioid use disorder by implicating the DNA methylation and gene expression.
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Dependencia de Heroína , Sitios de Carácter Cuantitativo , Metilación de ADN , Proteínas de Transporte de Dopamina a través de la Membrana Plasmática , Heroína , Dependencia de Heroína/tratamiento farmacológico , Dependencia de Heroína/genética , Humanos , Metadona/uso terapéutico , Polimorfismo de Nucleótido Simple/genéticaRESUMEN
A variety of mammalian or insect behaviors rely on the recognition of relevant odor stimuli. The olfactory system detects and translates complex olfactory stimuli (odors) through the unique and reproducible dynamic ensembles of neuronal activities. This process is involved in various types of neurons of olfactory parts, thereby encoding olfactory information or predicting progression in some neuropsychiatric diseases. In this paper, we constructed a biomimetic model including olfactory sensing system and olfactory bulb processing system to map olfactory-associated ensembles of neuronal activity. The olfactory receptor neurons (ORNs) and olfactory bulb (OB) neurons were primarily cultured and the immunofluorescence images were performed to identify the types of neurons. Diacetyl solution was used as an odor stimulus, and the spike bursts and random spike firing patterns of concentration-dependent excitatory responses were obtained from the ORNs network. The spike waveform and feature parameters were extracted including the spike number and interval in per burst to program the stimulation unit and sequences. The sequences containing odor information were applied to the OB neuronal network for the simulation of the primary olfactory processing. The response pattern and change rule of the OB neuronal network were consistent with the OB neurons affected by the neurotransmitter, which is the carrier of olfactory information transmission in vivo. This biomimetic integrated olfactory sensory and processing system can serve as a novel model for studying the physiological and pathological mechanisms of olfaction, and the pharmacological application in vitro.
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Técnicas Biosensibles , Olfato , Animales , Biomimética , Dispositivos Laboratorio en un Chip , Odorantes , Bulbo Olfatorio , Vías OlfatoriasRESUMEN
The biological olfactory and gustation system can discriminate thousands of odor and taste substances with high sensitivity and specificity, specific receptor proteins play an important role in this process. This study used the human neuroblastoma SH-SY5Y cell line endogenously expressing the human bitter receptor, T2R16. Meanwhile, an olfactory receptor, ODR-10, was transfected on the plasma membrane of SH-SY5Y cells. T2R16 could specifically respond to bitter compounds with the structure of ß-glucopyranosides by activation of G protein coupled receptors (GPCRs) causing cell morphologic changes, which could be monitored using a cell-impedance sensor. ODR-10 could specifically respond to diacetyl by changing the extracellular potential of the cells, the resopnse was recorded by a microelectrode array (MEA). The cell index (CI) value and firing rates were extracted from the signals as the biosensor response characteristics. The results with the sensors indicated a dose-dependent response within a defined concentration range. Moreover, this cell-impedance biosensor enabled quick toxicity detection of salicin when the concentration was ≥6â¯mM. In conclusion, the biomimetic sensors integrated olfaction, gustation and toxicity detection using the same cell, and has showed great potential for use in both basic research and practical applications.
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Bioingeniería , Biomimética , Técnicas Biosensibles , Olfato , Gusto , Alcoholes Bencílicos/análisis , Calcio/metabolismo , Diferenciación Celular/efectos de los fármacos , Línea Celular Tumoral , Diacetil/análisis , Glucósidos/análisis , Humanos , Microelectrodos , Odorantes/análisis , Receptores Acoplados a Proteínas G/metabolismo , Receptores Odorantes/metabolismo , Tretinoina/farmacologíaRESUMEN
Alzheimer's disease (AD) is a chronic central neurodegenerative disease. The pathological features of AD are the extracellular deposition of senile plaques formed by amyloid-ß oligomers (AßOs) and the intracellular accumulation of neurofibrillary tangles. However, due to the lack of effective method and experimental models to study the cognitive decline, communication at cell resolution and the implementation of interventions, the diagnosis and treatment on AD still progress slowly. In this paper, we established a pathological model of AD in vitro based on AßOs-induced hippocampal neuronal network chip for multi-site dynamic analysis of the neuronal electrical activity and network connection. The multiple characteristic parameters, including positive and negative spike intervals, firing rate and peak-to-peak values, were extracted through the analysis of spike signals, and two firing patterns from the interneurons and pyramidal neurons were recorded. The spatial firing patterns mapping and cross-correlation between channels were performed to validate the degeneration of neuronal network connectivity. Moreover, an electrical stimulation with frequency at 40â¯Hz was exerted to preliminarily explore the therapeutic effect on the pathological model of AD. This neuronal network chip enables the implementation of AD models in vitro for studying basic mechanisms of neurodegeneration within networks and for the parallel testing of various potential therapies. It can be a novel technique in the research of AD pathological model in vitro.
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Enfermedad de Alzheimer/diagnóstico , Péptidos beta-Amiloides/aislamiento & purificación , Técnicas Biosensibles , Sistemas Microelectromecánicos/métodos , Péptidos beta-Amiloides/química , Estimulación Eléctrica , Electrólitos/química , Hipocampo/química , Hipocampo/efectos de la radiación , Humanos , Interneuronas/química , Interneuronas/efectos de la radiación , Dispositivos Laboratorio en un Chip , Red Nerviosa/química , Red Nerviosa/efectos de la radiación , Ovillos Neurofibrilares/química , Ovillos Neurofibrilares/efectos de la radiación , Células Piramidales/química , Células Piramidales/efectos de la radiaciónRESUMEN
The olfactory system is a natural biosensor since its peripheral olfactory sensory neurons (OSNs) respond to the external stimuli and transmit the signals to the olfactory bulb (OB) where they are integrated and processed. The axonal connections from the OSNs expressing about 1000 different types of odorant receptors are precisely organized and sorted out onto 1800 glomeruli in the OB, from which the olfactory information is delivered to and perceived by the central nervous system. This process is carried out with particularly high sensitivity, specificity and rapidity, which can be used for explosive detection. Biomimetic olfactory biosensors use various biological components from the olfactory system as sensing elements, possessing great commercial prospects. In this study, we utilized the genetically labeled murine M72 olfactory sensory neurons with the green fluorescent protein (GFP) as sensing components and obtained long-term in vivo electrophysiological recordings from the M72 OSNs by implanting the microelectrode arrays (MEAs) into the behaving mouse's OB. The electrophysiological responses showed high reliability, reproducibility and specificity for odor detection, and particularly, the high sensitivity for the detection of odorants that contain benzene rings. Furthermore, our results indicated that it can detect trinitrotoluene (TNT) in liquid at a concentration as low as 10-5M and can distinguish TNT from other chemicals with a similar structure. Thus our study demonstrated that the in vivo biomimetic olfactory system could provide novel approaches to enhancing the specificity and increasing working lifespan of olfactory biosensors capable of detecting explosives.
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Técnicas Biosensibles/instrumentación , Odorantes/análisis , Neuronas Receptoras Olfatorias/metabolismo , Receptores Odorantes/metabolismo , Animales , Técnicas Biosensibles/métodos , Electrodos Implantados , Diseño de Equipo , Sustancias Explosivas/análisis , Sustancias Explosivas/metabolismo , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Masculino , Ratones Transgénicos , Microelectrodos , Modelos Moleculares , Bulbo Olfatorio/citología , Bulbo Olfatorio/fisiología , Receptores Odorantes/genética , Trinitrotolueno/análisis , Trinitrotolueno/metabolismoRESUMEN
The perception of sour taste in mammals is important for its basic modality properties and avoiding toxic substances. We explore a biomimetic bioelectronic tongue, which integrate MEA (microelectrode array) and taste receptor cell for acid detection as a switch. However, the acid-sensing mechanism and coding of the taste receptor cells in the periphery is not well understood, with long-standing debate. Therefore, we firstly construct a Hodgkin-Huxley type mathematical model of whole-cell acid-sensing taste receptor cells based on the electrophysiologic patch clamp recordings with different acid sensitive receptor expressing and different acidic stimulations. ASICs and PKDL channels are two most promising candidates for acidic sensation. ASICs channels contribute to the On response, and PKDL channels coding the Offset stimulations respectively, which function as a pair for switch. Therefore, with the advantage of effective and noninvasive detection for MEA, a sour taste biosensor based on MEA and taste receptor cells was designed and established to detect sour response from the elementary acid sensitive taste receptor cells during and after stimulus. From simulation and extracelluar potential recordings, we found the biomimetic bioelectronic tongue was acid-sensitive, as acid stimulation pH decrease, the firing frequency significantly increase. Furthermore, this reliable and effective MEA based bioelectronic tongue functioned as a switch for stimulation On and Off. This study provided a powerful platform to recognize sour stimulation and help elucidate the sour taste sensation and coding mechanism.
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Técnicas Biosensibles/instrumentación , Nariz Electrónica , Papilas Gustativas/citología , Animales , Biomimética/instrumentación , Células Cultivadas , Diseño de Equipo , Femenino , Humanos , Microelectrodos , Ratas Sprague-Dawley , GustoRESUMEN
Ovarian hormones, including progesterone (P4) and 17 ß-estradiol (E2), have been shown to affect memory functions; however, the underlying mechanism whereby ovarian hormone replacement therapy may decrease the risk of Alzheimer's disease (AD) is currently unclear. The present study aimed to investigate the effects of P4 and E2 on spatial and learning memory in an ovariectomized rat model of AD. ß-amyloid (Aß) or saline were stereotaxically injected into the hippocampus of the rats and, after 1 day, ovariectomy or sham operations were performed. Subsequently, the rats were treated with P4 alone, E2 alone, or a combination of P4 and E2. Treatment with E2 and/or P4 was shown to improve the learning and memory functions of the rats, as demonstrated by the Morris water maze test. In addition, treatment with E2 and P4 was associated with increased expression levels of choline acetyltransferase and 5-hydroxytryptamine receptor 2A (5-HT2A), and decreased expression levels of the glial fibrillary acidic protein in the hippocampus of the rats. Furthermore, E2 and P4 treatment significantly attenuated neuronal cell apoptosis, as demonstrated by terminal deoxynucleotidyl transferase dUTP nick end labeling assays; thus suggesting that the ovarian hormones were able to protect against Aß-induced neuronal cell toxicity. The results of the present study suggested that the neuroprotective effects of P4 and E2 were associated with amelioration of the cholinergic deficit, suppression of apoptotic signals and astrogliosis, and upregulation of 5-HT2A expression levels. Therefore, hormone replacement therapy may be considered an effective strategy for the treatment of patients with cognitive disorders and neurodegenerative diseases.