RESUMEN
5-Hydroxytryptamine (5-HT) is an amine produced in both the mammary gland and the central nervous system. Tryptophan hydroxylase 1 (TPH1) catalyzes the conversion of 5-hydroxytryptophan (5-HTP) into l-tryptophan, which is then converted into 5-HT by monoamine-oxidase (MAO-A). In the mammary gland, 5-HT has been shown to have a variety of paracrine-autocrine actions, including suppressing lactation, controlling the destiny of mammary epithelial cells, and maintaining calcium homeostasis throughout the transition from pregnancy to lactation. To examine the effects of 5-HT on the composition of colostrum and milk, a total of 30 transition Guan Zhong dairy goats were intramuscularly injected with 5-HTP (1.0 mg/kg) every morning before feeding from 10 d before the projected parturition date to the day of parturition. The average number of days animals received injections was 8.2 ± 3.2 d. 5-HTP treatment increased serum 5-HT concentration from days 5 to 2 relative to parturition (P < 0.05), and decreased the casein concentration of colostrum (P < 0.05). In the in vitro experiment, mammary epithelial cells isolated from three individual goats' mammary glands were separately treated with 200 µM 5-HTP, 30 µM PCPA (the specific inhibitor of TPH1), or 200 µM 5-HTP + 50 µM SB269970 (the selective antagonist of 5-HTR7). The results showed that 200 µM 5-HTP inhibited the expression of ß-casein, downregulated the activity of the JAK2/ STAT5a signaling pathway, and promoted the apoptosis of goat mammary epithelial cells (GMECs) (P < 0.05). When GMECs were treated with 30 µM Four-chloro-dl-phenylalanine (PCPA), a specific inhibitor of 5-HT synthesis, the mRNA expression of STAT5a and the phosphorylated STAT5a protein level were upregulated. The 50 µM SB269970 treatment rescued the effects of 5-HTP on GMECs (P < 0.05). Taken together, the results indicated that 5-HTP exerted an inhibitory effect on ß-casein synthesis and a proapoptotic effect in GMECs via HTR7 and the JAK2/STAT5a axis.
5-Hydroxytryptamine (5-HT), which is produced in both the mammary gland and the central nervous system, is a recognized important regulator of mammary gland homeostasis. Casein is the major protein in the milk of mammals including cows, goats, and humans, and is a crucial source of high-quality amino acids for humans. In this study, prenatal intramuscular injection of 5-hydroxytryptophan (5-HTP), the precursor of 5-HT, not only increased the level of 5-HT in the serum of goats before delivery but also decreased the concentration of casein in colostrum. Furthermore, in goat mammary epithelial cells which are responsible for milk synthesis, it was found that 5-HTP blocked genes and signal pathways related to casein synthesis, and also promoted cell apoptosis. Additional results demonstrated that the type 7 5-HT receptor (HTR7) mediated the impacts of 5-HT, which provided a potential reliable target for improving milk quality.
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5-Hidroxitriptófano , Caseínas , Animales , Femenino , Embarazo , 5-Hidroxitriptófano/farmacología , 5-Hidroxitriptófano/metabolismo , Apoptosis , Caseínas/metabolismo , Células Epiteliales/metabolismo , Cabras/genética , Lactancia , Glándulas Mamarias Animales/metabolismo , Serotonina/farmacología , Serotonina/metabolismo , Factor de Transcripción STAT5/genética , Factor de Transcripción STAT5/metabolismo , Factor de Transcripción STAT5/farmacología , Receptores de Serotonina/metabolismoRESUMEN
BACKGROUND: Breast abscess during lactation is a severe complication of acute mastitis, which can lead to discomfort, high fever, breast fistula, sepsis, septic shock, breast damage, disease persistence and frequent hospitalization. Breast abscesses may also lead the mother to discontinue breastfeeding, thereby harming the infant's health. The predominant pathogenic bacteria are Staphylococcus aureus, Staphylococcus epidermidis and Streptococcus. The incidence of breastfeeding abscesses in breastfeeding women ranges between 4.0% and 11.0%. In cases of breast abscess, the rate of cessation of lactation is 41.0%. In instances of breast fistula, the rate of cessation of lactation is very high (66.7%). Furthermore, 50.0% of women with breast abscesses must be hospitalized and treated with intravenous antibiotics. Treatment includes antibiotics, abscess puncture and surgical incision and drainage. The patients suffer from stress, pain and easily induced breast scarring; the disease's progression is prolonged and recurrent, interfering with infant feeding. Consequently, it is crucial to discover an adequate cure. CASE SUMMARY: A 28-year-old woman with a breast abscess was treated with Gualou Xiaoyong decoction and painless breast opening manipulation 24 d after cesarean delivery. On the 2nd d of treatment, the patient's breast mass was significantly reduced, the pain was significantly reduced, and the general asthenia was improved. All conscious symptoms disappeared after 3 d, breast abscesses faded after 12 d of treatment, inflammation images disappeared after 27 d, and normal lactation images were restored. CONCLUSION: In treating breast abscesses during breastfeeding, the combination of Gualou Xiaoyong decoction and painless lactation provides a positive therapeutic impact. This disease's treatment offers the advantages of a short course of treatment, no need to discontinue breastfeeding and the ability to rapidly mitigate symptoms, which can be used as a reference in clinical practice.
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During mammary development, the transdifferentiation of mammary preadipocytes is one of the important sources for lactating mammary epithelial cells (MECs). However, there is limited knowledge about the mechanisms of dynamic regulation of transcriptome and genome-wide DNA methylation in the preadipocyte transdifferentiation process. Here, to gain more insight into these mechanisms, preadipocytes were isolated from adipose tissues from around the goat mammary gland (GM-preadipocytes). The GM-preadipocytes were cultured on Matrigel in conditioned media made from goat MECs to induce GM-preadipocyte-to-MEC transdifferentiation. The transdifferentiated GM-preadipocytes showed high abundance of keratin 18, which is a marker protein of MECs, and formed mammary acinar-like structures after 8 days of induction. Then, we performed transcriptome and DNA methylome profiling of the GM-preadipocytes and transdifferentiated GM-preadipocytes, respectively, and the differentially expressed genes and differentially methylated genes that play underlying roles in the process of transdifferentiation were obtained. Subsequently, we identified the candidate transcription factors in regulating the GM-preadipocyte-to-MEC transdifferentiation by transcription factor-binding motif enrichment analysis of differentially expressed genes and differentially methylated genes. Meanwhile, the secretory proteome of GM-preadipocytes cultured in conditioned media was also detected. By integrating the transcriptome, DNA methylome, and proteome, three candidate genes, four proteins, and several epigenetic regulatory axes were further identified, which are involved in regulation of the cell cycle, cell polarity establishment, cell adhesion, cell reprogramming, and adipocyte plasticity. These findings provide novel insights into the molecular mechanism of preadipocyte transdifferentiation and mammary development.
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Metilación de ADN , Lactancia , Animales , Femenino , Medios de Cultivo Condicionados , Células Epiteliales/metabolismo , Perfilación de la Expresión Génica , Cabras , Lactancia/genética , Glándulas Mamarias Animales , Proteoma/metabolismo , Transcriptoma , Adipocitos/metabolismoRESUMEN
Circular RNAs (circRNAs), characterized as single-stranded closed circular RNA molecules, have been established to exert pivotal functions in various biological or pathological processes. Nonetheless, the effects and underlying mechanisms concerning circRNAs on the aging and aging-related diseases remain elusive. We herein compared the expression patterns of circRNAs in young and senescent mouse embryonic fibroblasts (MEFs), and uncovered that circRNF169 was dramatically up-regulated in senescent MEFs compared with that in young MEFs. Therefore, we further digged into the role and potential mechanisms of circRNF169 in the senescence of MEFs. The results of senescence-associate-ß-galactosidase staining and BrdU incorporation assay showed that silencing of circRNF169 significantly delayed MEFs senescence and promoted cell proliferation, while ectopic expression of circRNF169 exhibited the opposite effects. Moreover, the dual-luciferase reporter assay confirmed that circRNF169 acted as an endogenous miR-30c-5p sponge, which accelerated cellular senescence by sequestering and inhibiting miR-30c-5p activity. Taken together, our results suggested that circRNF169 exerted a crucial role in cellular senescence through sponging miR-30c-5p and represented a promising target for aging intervention.
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Senescencia Celular , MicroARNs , ARN Circular , Animales , Proliferación Celular/genética , Senescencia Celular/genética , Fibroblastos/metabolismo , Ratones , MicroARNs/genética , MicroARNs/fisiología , ARN Circular/genética , ARN Circular/fisiologíaRESUMEN
BACKGROUND: Depression is a common psychiatric disorder associated with defects in GABAergic (gamma-aminobutyric acidergic) neurotransmission. α-Dystroglycan (α-DG), a cell adhesion molecule known to be essential for skeletal muscle integrity, is also present at inhibitory synapses in the central nervous system and forms a structural element in certain synapses. However, the role of α-DG in the regulation of depressive-like behaviors remains largely unknown. METHODS: Depressive-like behaviors were induced by chronic social defeat stress in adult male mice. Surface protein was extracted by a biotin kit, and the expression of protein was detected by Western blotting. Intrahippocampal microinjection of the lentivirus or adeno-associated virus or agrin intervention was carried out using a stereotaxic instrument and followed by behavioral tests. Miniature inhibitory postsynaptic currents were recorded by whole-cell patch-clamp techniques. RESULTS: The expression of α-DG and glycosylated α-DG in the ventral hippocampus was significantly lower in chronic social defeat stress-susceptible male mice than in control mice, accompanied by a decreased surface expression of GABAA receptor γ2 subunit and reduced GABAergic neurotransmission. RNA interference-mediated knockdown of Dag1 increased the susceptibility of mice to subthreshold stress. Both in vivo administration of agrin and overexpression of like-acetylglucosaminyltransferase ameliorated depressive-like behaviors and restored the decrease in surface expression of GABAA receptor γ2 subunit and the amplitude of miniature inhibitory postsynaptic currents in chronic social defeat stress-exposed mice. CONCLUSIONS: Our findings demonstrate that glycosylated α-DG plays a role in the pathophysiological process of depressive-like behaviors by regulating the surface expression of GABAA receptor γ2 subunit and GABAergic neurotransmission in the ventral hippocampus.
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Distroglicanos , Receptores de GABA-A , Agrina/metabolismo , Animales , Distroglicanos/metabolismo , Hipocampo/metabolismo , Humanos , Masculino , Ratones , Receptores de GABA/metabolismo , Receptores de GABA-A/metabolismo , Ácido gamma-Aminobutírico/metabolismoRESUMEN
The cytosolic isozyme of phosphoenolpyruvate carboxykinase (PCK1) was the first rate-limiting enzyme in the gluconeogenesis pathway, which exerted a critical role in maintaining the blood glucose levels. PCK1 has been established to be involved in various physiological and pathological processes, including glucose metabolism, lipid metabolism, diabetes, and tumorigenesis. Nonetheless, the association of PCK1 with aging process and the detailed underlying mechanisms of PCK1 on aging are still far to be elucidated. Hence, we herein constructed the PCK1-deficient (pck1Δ) and PCK1 overexpression (PCK1 OE) Saccharomyces cerevisiae. The results unveiled that PCK1 deficiency significantly shortened the replicative lifespan (RLS) in the S. cerevisiae, while overexpression of PCK1 prolonged the RLS. Additionally, we noted that the ROS level was significantly enhanced in PCK1-deficient strain and decreased in PCK1 OE strain. Then, a high throughput analysis by deep sequencing was performed in the pck1Δ and wild-type strains, in an attempt to shed light on the effect of PCK1 on the lifespan of aging process. The data showed that the most downregulated mRNAs were enriched in the regulatory pathways of glucose metabolism. Fascinatingly, among the differentially expressed mRNAs, PFK1 was one of the most upregulated genes, which was involved in the glycolysis process and ROS generation. Thus, we further constructed the pfk1Δpck1Δ strain by deletion of PFK1 in the PCK1-deficient strain. The results unraveled that pfk1Δpck1Δ strain significantly suppressed the ROS level and restored the RLS of pck1Δ strain. Taken together, our data suggested that PCK1 deficiency enhanced the ROS level and shortened the RLS of S. cerevisiae via PFK1.
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Regulación Enzimológica de la Expresión Génica , Regulación Fúngica de la Expresión Génica , Fosfoenolpiruvato Carboxiquinasa (ATP) , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae , Fosfoenolpiruvato Carboxiquinasa (ATP)/deficiencia , Fosfoenolpiruvato Carboxiquinasa (ATP)/metabolismo , Saccharomyces cerevisiae/enzimología , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismoRESUMEN
BACKGROUND: Pregnancy in the setting of systemic lupus erythematosus can worsen the condition from the stable to active stage, with quality of life and fertility desire being particular concerns. Pregnancy in the active stage of systemic lupus erythematosus (ASLE), although rare and complicated to manage, can be treated favorably with immunotherapies ifs used properly. Here we report such a success case. CASE SUMMARY: A 31-year-old primigravida patient, diagnosed with SLE seven years ago, was induced ASLE after a cold at 21 + weeks. The patient's vital signs on presentation were normal. Her laboratory exam was remarkable for significant proteinuria, liver and renal dysfunction, and low C3 and C4 levels. Infectious work-up was negative. The patient was diagnosed with ASLE. She was given immunosuppressive agents (methylprednisolone, gamma globulin and azathioprine etc.) and plasma adsorption therapy, monitoring blood pressure every 8 h, fetal heart rate twice a day, and liver and renal function at least twice a week. Successful maternal and fetal outcomes are presented here. CONCLUSION: Child-bearing in ASLE has become more promising, even for this difficult case of ASLE with multiple organ damage. Thorough antepartum counseling, cautious maternal-fetal monitoring, and multi-organ function monitoring by multidisciplinary specialties are keys to favorable pregnancy outcomes.
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Cocaine is a strong central nervous system stimulant, which can induce drug addiction. Previous studies have reported that cocaine-induced autophagy is involved in neuroinflammation and cell death. However, the role of autophagy in psychomotor sensitivity to cocaine has not been explored. Here, we reported that D1 receptor -CaMKII-AMPK-FoxO3a signaling pathway was involved in acute cocaine application-induced autophagy in the nucleus accumbens (NAc) both in vitro and in vivo. Furthermore, we found that knockdown of the ATG5 gene in the NAc augmented behavioral response to cocaine, and induction of autophagy in the NAc with rapamycin attenuated cocaine-induced behavioral response, which was coincident with the alterations of dendritic spine density in neurons of NAc. These results suggest that cocaine exposure leads to the induction of autophagy, which is a protective mechanism against behavioral response to cocaine of male mice.
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Proteínas Quinasas Activadas por AMP/metabolismo , Autofagia/efectos de los fármacos , Conducta Animal/efectos de los fármacos , Trastornos Relacionados con Cocaína/prevención & control , Cocaína/farmacología , Núcleo Accumbens/metabolismo , Animales , Trastornos Relacionados con Cocaína/metabolismo , Trastornos Relacionados con Cocaína/fisiopatología , Modelos Animales de Enfermedad , Inhibidores de Captación de Dopamina/farmacología , Masculino , Ratones , Ratones Endogámicos C57BL , Núcleo Accumbens/efectos de los fármacosRESUMEN
Cocaine is a common abused drug that can induce abnormal synaptic and immune responses in the central nervous system (CNS). High mobility group box 1 (HMGB1) is one kind of inflammatory molecules that is expressed both on neurons and immune cells. Previous studies of HMGB1 in the CNS have largely focused on immune function, and the role of HMGB1 in neurons and cocaine addiction remains unknown. Here, we show that cocaine exposure induced the translocation and release of HMGB1 in the nucleus accumbens (NAc) neurons. Gain and loss of HMGB1 in the NAc bidirectionally regulate cocaine-induced conditioned place preference. From the nucleus to the cytosol, HMGB1 binds to glutamate receptor subunits (GluA2/GluN2B) on the membrane, which regulates cocaine-induced synaptic adaptation and the formation of cocaine-related memory. These data unveil the role of HMGB1 in neurons and provide the evidence for the HMGB1 involvement in drug addiction.
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Trastornos Relacionados con Cocaína/genética , Proteína HMGB1/genética , Memoria/efectos de los fármacos , Neuronas/efectos de los fármacos , Núcleo Accumbens/efectos de los fármacos , Recompensa , Animales , Cocaína/farmacología , Trastornos Relacionados con Cocaína/fisiopatología , Modelos Animales de Enfermedad , Masculino , Núcleo Accumbens/fisiopatología , Ratas , Ratas Sprague-DawleyRESUMEN
Aging is a multifactorial process characterized by the progressive deterioration of physiological functions. Among the multiple molecular mechanisms, microRNAs (miRNAs) have increasingly been implicated in the regulation of Aging process. However, the contribution of miRNAs to physiological Aging and the underlying mechanisms remain elusive. We herein performed high-throughput analysis using miRNA and mRNA microarray in the physiological Aging mouse, attempted to deepen into the understanding of the effects of miRNAs on Aging process at the "network" level. The data showed that various p53 responsive miRNAs, including miR-124, miR-34a and miR-29a/b/c, were up-regulated in Aging mouse compared with that in Young mouse. Further investigation unraveled that similar as miR-34a and miR-29, miR-124 significantly promoted cellular senescence. As expected, mRNA microarray and gene co-expression network analysis unveiled that the most down-regulated mRNAs were enriched in the regulatory pathways of cell proliferation. Fascinatingly, among these down-regulated mRNAs, Ccna2 stood out as a common target of several p53 responsive miRNAs (miR-124 and miR-29), which functioned as the antagonist of p21 in cell cycle regulation. Silencing of Ccna2 remarkably triggered the cellular senescence, while Ccna2 overexpression delayed cellular senescence and significantly reversed the senescence-induction effect of miR-124 and miR-29. Moreover, these p53 responsive miRNAs were significantly up-regulated during the senescence process of p21-deficient cells; overexpression of p53 responsive miRNAs or knockdown of Ccna2 evidently accelerated the cellular senescence in the absence of p21. Taken together, our data suggested that the p53/miRNAs/Ccna2 pathway might serve as a novel senescence modulator independent of p53/p21 pathway.
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Senescencia Celular , Ciclina A2/metabolismo , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , MicroARNs/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Animales , Células Cultivadas , Senescencia Celular/genética , Ciclina A2/deficiencia , Ciclina A2/genética , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , MicroARNs/genética , Células 3T3 NIH , Proteína p53 Supresora de Tumor/genéticaRESUMEN
ß-Catenin has been implicated in major depressive disorder (MDD), which is associated with synaptic plasticity and dendritic arborization. MicroRNAs (miRNA) are small noncoding RNAs containing about 22 nucleotides and involved in a variety of physiological and pathophysiological process, but their roles in MDD remain largely unknown. Here, we investigated the expression and function of miRNAs in the mouse model of chronic social defeat stress (CSDS). The regulation of ß-catenin by selected miRNA was validated by silico prediction, target gene luciferase reporter assay, and transfection experiment in neurons. We demonstrated that the levels of miR-214-3p, which targets ß-catenin transcripts were significantly increased in the medial prefrontal cortex (mPFC) of CSDS mice. Antagomir-214-3p, a neutralizing inhibitor of miR-214-3p, increased the levels of ß-catenin and reversed the depressive-like behavior in CSDS mice. Meanwhile, antagomir-214-3p increased the amplitude of miniature excitatory postsynaptic current (mEPSC) and the number of dendritic spines in mPFC of CSDS mice, which may be related to the elevated expression of cldn1. Furthermore, intranasal administered antagomir-214-3p also significantly increased the level of ß-catenin and reversed the depressive-like behaviors in CSDS mice. These results may represent a new therapeutic target for MDD.
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Depresión/fisiopatología , MicroARNs/fisiología , Estrés Psicológico/fisiopatología , beta Catenina/fisiología , Administración Intranasal , Animales , Antagomirs/administración & dosificación , Claudina-1/genética , Espinas Dendríticas/efectos de los fármacos , Espinas Dendríticas/fisiología , Depresión/etiología , Depresión/genética , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Regulación de la Expresión Génica , Hipocampo/efectos de los fármacos , Hipocampo/fisiopatología , Masculino , Ratones Endogámicos C57BL , MicroARNs/antagonistas & inhibidores , MicroARNs/genética , Corteza Prefrontal/efectos de los fármacos , Corteza Prefrontal/fisiopatología , Estrés Psicológico/genética , beta Catenina/genéticaRESUMEN
BACKGROUND: Benzodiazepines (BZDs) have been used to treat anxiety disorders for more than five decades as the allosteric modulator of the gamma-aminobutyric acid A receptor (GABAAR). Little is known about other mechanisms of BZDs. Here, we describe how the rapid stabilization of postsynaptic GABAAR is essential and sufficient for the anxiolytic effect of BZDs via a palmitoylation-dependent mechanism. METHODS: Palmitoylated proteins in the basolateral amygdala (BLA) of rats with different anxious states were assessed by a biotin exchange protocol. Both pharmacological and genetic approaches were used to investigate the role of palmitoylation in anxiety behavior. Electrophysiological recording, reverse transcription polymerase chain reaction, Western blotting, and coimmunoprecipitation were used to investigate the mechanisms. RESULTS: Highly anxious rats were accompanied by the deficiency of gephyrin palmitoylation and decreased the synaptic function of GABAAR in the BLA. We then identified that the dysfunction of DHHC12, a palmitoyl acyltransferase that specifically palmitoylates gephyrin, contributed to the high-anxious state. Furthermore, diazepam, as an anxiolytic drug targeting GABAARs, was found to increase gephyrin palmitoylation in the BLA via a GABAAR-dependent manner to activate DHHC12. The anxiolytic effect of diazepam was nearly abolished by the DHHC12 knockdown. Specifically, similar to the effect of BZD, the overexpression of DHHC12 in the BLA exerted a significant anxiolytic action, which was prevented by flumazenil. CONCLUSIONS: Our results support the view that the strength of inhibitory synapse was controlled by gephyrin palmitoylation in vivo and proposes a previously unknown palmitoylation-centered mode of BZD's action.
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Ansiedad/metabolismo , Complejo Nuclear Basolateral/metabolismo , Benzodiazepinas/farmacología , Proteínas de la Membrana/metabolismo , Aciltransferasas/genética , Aciltransferasas/metabolismo , Animales , Ansiolíticos/farmacología , Diazepam/farmacología , Flumazenil/farmacología , Técnicas de Silenciamiento del Gen , Lipoilación , Masculino , Ratas , Receptores de GABA-A/metabolismo , Receptores de GABA-A/fisiologíaRESUMEN
Senescence-associated secretory phenotype (SASP) is often a concomitant result of cell senescence, embodied by the enhanced function of secretion. The SASP factors secreted by senescent cells include cytokines, proteases and chemokines, etc, which can exert great influence on local as well as systemic environment and participate in the process of cell senescence, immunoregulation, angiogenesis, cell proliferation and tumor invasion, etc. Relative to the abundance of SASP models in human cells, the in vitro SASP model derived from mouse cells is scarce at present. Therefore, the study aimed to establish a mouse SASP model to facilitate the research in the field. With this objective, we treated the INK4a-deficient mouse NIH-3T3 cells and the wildtype mouse embryonic fibroblasts (MEF) respectively with mitomycin C (MMC), an anticarcinoma drug which could induce DNA damage. The occurring of cell senescence was evaluated by cell morphology, ß-gal staining, integration ratio of EdU and Western blot. Quantitative RT-PCR and ELISA were used to detect the expression and secretion of SASP factors, respectively. The results showed that, 8 days after the treatment of NIH-3T3 cells with MMC (1 µg/mL) for 12 h or 24 h, the cells became enlarged and the ratios of ß-gal-positive (blue-stained) cells significantly increased, up to 77.4% and 90.4%, respectively. Meanwhile, the expression of P21 protein increased and the integration ratios of EdU significantly decreased (P < 0.01). Quantitative RT-PCR detection showed that the mRNA levels of several SASP genes, including IL-6, TNF-α, IL-1α and IL-1ß increased evidently. ELISA detection further observed an enhanced secretion of IL-6 (P < 0.01). On the contrary, although wildtype MEF could also be induced into senescence by MMC treatment for 12 h or 24 h, embodied by the enlarged cell volume, increased ratios of ß-gal-positive cells (up to 71.7% and 80.2%, respectively) and enhanced expression of P21 protein, the secretion of IL-6 displayed no significant change. Our study indicated that, although MMC could induce senescence in both mouse NIH-3T3 cells and wildtype MEF, only senescent NIH-3T3 cells displayed the canonical SASP phenomena. Current study suggested that senescent NIH-3T3 cells might be an appropriate in vitro SASP model of mouse cells.
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Senescencia Celular/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Mitomicina/farmacología , Animales , Proliferación Celular , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/genética , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Citocinas/genética , Citocinas/metabolismo , Daño del ADN , Interleucina-6/metabolismo , Ratones , Células 3T3 NIH , FenotipoRESUMEN
Aquaporin-4 (AQP-4) is the predominant water channel in the brain and primarily expressed in astrocytes. Astrocytes have been generally believed to play important roles in regulating synaptic plasticity and information processing. A growing number of evidence shows that AQP-4 plays a potential role in the regulation of astrocyte function. However, little is known about the function of AQP-4 for synaptic plasticity in the hippocampus. Therefore, we evaluated long-term depression (LTD) in the hippocampus and the extinction of fear memory of AQP-4 knockout (KO) and wild-type (WT) mice. We found that AQP-4 deficiency facilitated fear memory extinction and NMDA receptors (NMDARs)-dependent LTD in the CA3-CA1 pathway. Furthermore, AQP-4 deficiency selectively increased GluN2B-NMDAR-mediated excitatory postsynaptic currents (EPSCs). The excessive activation of extrasynaptic GluN2B-NMDAR contributed to the facilitation of NMDAR-dependent LTD and enhancement of fear memory extinction in AQP-4 KO mice. Thus, it appears that AQP-4 may be a potential target for intervention in fear memory extinction. This article is part of the Special Issue entitled 'Ionotropic glutamate receptors'.
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Acuaporina 4/fisiología , Extinción Psicológica/fisiología , Miedo/fisiología , Hipocampo/fisiología , Depresión Sináptica a Largo Plazo , Receptores de N-Metil-D-Aspartato/fisiología , Animales , Acuaporina 4/genética , Sistema de Señalización de MAP Quinasas , Masculino , Ratones , Ratones NoqueadosRESUMEN
MicroRNA-194 (miR-194), a typical p53 responsive miRNA, serves as a tumor suppressor similar as p53, and has been demonstrated to play an anti-proliferation role in various human cancers. In spite of the pivotal role of p53 during aging process, the knowledge of miR-194's contribution to cellular senescence is limited. We herein sought to explore the role of miR-194 in the replicative senescence and stress-induced senescence of mouse embryonic fibroblasts. Our results unraveled that, compared to young cells, miR-194 is highly expressed in senescent cells, and extra expression of miR-194 significantly triggers the replicative senescence of MEFs and H2 O2 -induced senescence of NIH/3T3 cells, while inhibition of miR-194 exhibited the opposite effect. We further unveiled that DNMT3A was a direct and authentic target of miR-194, which has been reported to be closely associated with cellular senescence. Taken together, our data suggest that miR-194 may significantly promote the development of cellular senescence in mouse embryonic fibroblasts, which potentially occurs through inhibiting the DNMT3A expression.
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Senescencia Celular/fisiología , Fibroblastos/fisiología , MicroARNs/biosíntesis , MicroARNs/genética , Animales , Células HEK293 , Células Endoteliales de la Vena Umbilical Humana/fisiología , Humanos , Ratones , Ratones Endogámicos C57BL , Células 3T3 NIHRESUMEN
MicroRNAs are a large class of tiny noncoding RNAs, which have emerged as critical regulators of gene expression, and thus are involved in multiple cellular processes, including cellular senescence. MicroRNA-33 has previously been established to exert crucial effect on cell proliferation, lipid metabolism and cholesterol metabolism. Nonetheless, the association between microRNA-33 and cellular senescence and its underlying molecular mechanism are far to be elucidated. The present study has attempted to probe into the effect of microRNA-33 on MEFs senescence. Our data unveiled that microRNA-33 was dramatically down-regulated in senescent MEFs compared to the young MEFs, and ectopic expression of microRNA-33 promoted MEFs senescence, while knock-down of microRNA-33 exhibited a protective effect against senescence phenotype. Moreover, we verified CDK6 as a direct target of microRNA-33 in mouse. Silencing of CDK6 induced the premature senescence phenotype of MEFs similarly as microRNA-33, while enforced expression of CDK6 significantly reverse the senescence-induction effect of microRNA-33. Taken together, our results suggested that microRNA-33 enhanced the replicative senescence of MEFs potentially by suppressing CDK6 expression.
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Senescencia Celular/fisiología , Quinasa 6 Dependiente de la Ciclina/metabolismo , Embrión de Mamíferos/fisiología , Fibroblastos/citología , Fibroblastos/fisiología , MicroARNs/metabolismo , Animales , Proliferación Celular/fisiología , Células Cultivadas , Embrión de Mamíferos/citología , Masculino , Ratones , Ratones Endogámicos C57BLRESUMEN
Chinese herbal medicine (CHM), as the largest application category of traditional Chinese medicine (TCM), is widely accepted among cancer patients in China. Herbal slice (HS) and Chinese patent drug (CPD) are commonly used CHM in China. This study aimed to investigate the utilization of CHM among clinicians and cancer patients in central China. Five hundred and twenty-five patients and 165 clinicians in 35 comprehensive hospitals in central China were asked to complete an anonymous questionnaire that was designed to evaluate the use of CHM. The results showed that 90.74% clinicians and 72.24% cancer patients used CHM during cancer treatment. The educational backgrounds of the clinicians and the age, education level, annual income, and cancer stage of the cancer patients were related to use of CHM. More than 90% clinicians and cancer patients had used CPD. Comparatively, the percentage of HS use was 10% lower than that of CPD use among clinicians and cancer patients. More clinicians preferred to use CHM after surgery than cancer patients did (20.41% vs. 5.37%). Enhancing physical fitness and improving performance status were regarded as the most potential effect of CHM on cancer treatment (85.71% among clinicians and 94.07% among cancer patients), in comparison with directly killing tumor cells (24.49% among clinicians and 31.36% among patients). As for refusal reasons, imprecise efficacy was the unanimous (100%) reason for clinicians' rejection of CHM, and 95.58% patients objected to using CHM also for this reason. Furthermore, the side effects of CHM were more concerned by clinicians than by patients (33.33% vs. 15.81%). In conclusion, our survey revealed that CHM was popularly accepted by clinicians and cancer patients in central China. The reasons of use and rejection of CHM were different between clinicians and cancer patients.
Asunto(s)
Medicina de Hierbas , Hospitales , Cuerpo Médico de Hospitales , Medicina Tradicional China , Neoplasias/terapia , Adulto , China , Femenino , Humanos , Masculino , Oncología Médica , Persona de Mediana Edad , Encuestas y Cuestionarios , Recursos HumanosRESUMEN
The recombinant adenoviruses which can overexpress fosb and Delta2Deltafosb proteins were produced. Primary calvarial osteoblasts from new born CD1 mice were infected with those adenoviruses by different proportion of Ad-Delta2Deltafosb and induced by 5 mmol/L beta-glycerophosphate, 50 microg/ml ascorbic acid and 10 nmol/L dexamethasone for 23 days and 29 days, respectively. The results by Alizarin Red Staining showed that the primary osteoblasts overexpressing Delta2Deltafosb protein formed more nodus than the control cells and the nodus in overexpressing Delta2Deltafosb protein osteoblasts were smaller than those in control cells on day 23. Differentiation of the primary osteoblasts overexpressing Delta2Deltafosb protein increased more than that of the control cells on day 29. It is an evidence for the reason that bone density of Delta2Deltafosb transgenic mice was markedly increased throughout the skeleton in new born and mature mice.