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BACKGROUND: Hypodermin A (HA) is a serine esterase that degrades complement, a key element of the innate immune system. Immunosuppressive properties of HA have previously been studied in vitro. However, such properties have not been fully demonstrated in vivo. The aim of this study was to evaluate the effect of HA in inhibiting allograft rejection in an HA transgenic mouse model. METHODS: FVB (HA transgenic mice or wild-type mice) to BALB/c mice skin transplantation model were used. Skin grafts were analyzed by histology, immunohistochemistry, and Western blotting. RESULTS: HA overexpression resulted in significantly prolonged skin allograft survival. Histologic changes in the skin allografts paralleled the gross appearance of rejection. ELISA and Western blotting showed that HA significantly reduced the content of complement C3 and C9 in HA skin allografts. The expressions of CD4, B7-2, and MHC class II were all significantly suppressed in HA skin allografts compared with the control group. CONCLUSIONS: These findings suggest that HA effectively prolongs skin allograft survival. The study results provide insight into a promising strategy to improve the survival of grafts in humans.
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Terapia Genética/métodos , Rechazo de Injerto/prevención & control , Supervivencia de Injerto/inmunología , Serina Endopeptidasas/inmunología , Trasplante de Piel , Animales , Biomarcadores/metabolismo , Western Blotting , Ensayo de Inmunoadsorción Enzimática , Femenino , Rechazo de Injerto/enzimología , Rechazo de Injerto/genética , Rechazo de Injerto/inmunología , Supervivencia de Injerto/genética , Inmunohistoquímica , Ratones , Ratones Endogámicos BALB C , Ratones Transgénicos , Serina Endopeptidasas/genética , Serina Endopeptidasas/metabolismo , Trasplante Homólogo , Resultado del Tratamiento , Regulación hacia ArribaRESUMEN
The mechanisms of fetal semi-allograft acceptance by the mother's immune system have been the target of many immunological studies. Early pregnancy factor (EPF) is a molecule present in the serum of pregnant mammals soon after conception that has been reported to have immunomodulatory effects. In the present study, we aimed to determine whether immune cells such as CD4+CD25+ regulatory T cells (Tregs) are involved in the suppressive mechanism of EPF. Accordingly, CD4+CD25- T cells were isolated from spleens of female C57BL/6 mice and stimulated with anti-CD3 antibody, anti-CD28 antibody and IL-2 in the presence or absence of EPF. Flow cytometry was used to analyze the differentiation of CD4+CD25- T cells to CD4+CD25+ Tregs. We thus found a remarkable rise in the Treg ratio in the EPF-treated cells. Higher mRNA and protein levels of fork head box P3 (Foxp3), a marker of the Treg lineage, were also observed in cells treated with EPF. Furthermore, the effect of EPF on Treg immunosuppressive capacity was evaluated. EPF treatment induced the expression of interleukin-10 and transforming growth factor ß1 in Tregs. The suppressive capacity of Tregs was further measured by their capability to inhibit T cell receptor-mediated proliferation of CD4+CD25- T cells. We thus found that EPF exposure can enhance the immunosuppressive functions of Tregs. Overall, our data suggest that EPF induces the differentiation of Tregs and increases their immunosuppressive activities, which might be an important mechanism to inhibit immune responses during pregnancy.
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Antígenos CD4/metabolismo , Chaperonina 10/farmacología , Subunidad alfa del Receptor de Interleucina-2/metabolismo , Proteínas Gestacionales/farmacología , Factores Supresores Inmunológicos/farmacología , Linfocitos T Reguladores/metabolismo , Animales , Femenino , Factores de Transcripción Forkhead/metabolismo , Humanos , Terapia de Inmunosupresión , Ratones Endogámicos C57BL , Receptores de Antígenos de Linfocitos T/metabolismo , Linfocitos T Reguladores/efectos de los fármacosRESUMEN
Studies on the bench and at bedside have demonstrated that the process of epileptogenesis is involved in neuroinflammatory responses. As the receptor of proinflammatory cytokine IL-1ß, IL-1ß type 1 receptor (IL-1R1) is reported to express abundantly in the endothelial cells in epileptic brains, which is deemed to be implicated in the epileptogenic process. However, whether and how endothelial IL-1R1 modulates neuroinflammatory responses in the pathological process of epileptic seizures and/or status epilepticus (SE) remains obscure. Here, we indicated endothelial IL-1R1 is involved in neuroinflammation, facilitating epilepsy progress via Nrf2/HO-1/NLRP3. In vitro, we observed upregulation of inflammatory cytokines in co-culture model under IL-1ß challenge, as well as in BV2 cells after stimulation with conditional medium (CM) from IL-1ß-stimulated bEnd.3 cells. In vivo, mice with conditional knockout of endothelial IL-1R1 (IL-1R1-CKO) were generated by hybrid IL-1R1flox/flox mice with Tek-Cre mice. IL-1R1-CKO reduced seizure susceptibility in kainic acid (KA)-induced SE model. In addition, IL-1R1-CKO KA mice exhibited lessened hippocampal neuroinflammation, mitigated neuronal damage, and decreased abnormal neurogenesis. In cognitive behavioral tests, IL-1R1-CKO KA mice presented improvement in learning and memory. Furthermore, we also indicated blockage of endothelial IL-1R1 downregulated the expressions of Nrf2/HO-1/NLRP3 pathway-related proteins. Nrf2-siRNA reversed the downregulation of HO-1, NLRP3, caspase-1, and IL-1ß. These results demonstrated CKO of endothelial IL-1R1 reduces seizure susceptibility and attenuates SE-related neurobehavioral damage by suppressing hippocampal neuroinflammation via Nrf2/HO-1/NLRP3.
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Purpose: A novel CYPA-targeted, SiO2 encapsulated Au star@AgAu yolk-shell nanostructure (YSNS) was synthesized and used for ovarian cancer early diagnosis and therapy. Methods: Diverse spectroscopic and microscopic methods were utilized to investigate the pattern of the yolk-shell nanostructure. In addition, in vitro and in vivo experiments were carried out. Results: It can be found that the ratio of HAuCl4 and AgNO3 played a critical role in the constitution of the yolk-shell nanostructure. The as-prepared yolk-shell nanostructure showed excellent SERS performance, which could be utilized as SERS substrate for specific sensitivity analysis of ovarian cancer markers cyclophilin A (CYPA) with detectable limit of 7.76*10-10 µg/mL. In addition, the as-prepared yolk-shell nanostructure possessed outstanding photothermal performance, which could be used as photothermal agent for ovarian cancer therapy. Experiments in vitro and in vivo proved that the as-prepared yolk-shell nanostructures are ideal candidate for early diagnosis and therapy for ovarian cancer in one platform. Conclusion: This work holds promise to offer a new method for the detection and therapy of ovarian cancer in the early stage.
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Nanoestructuras , Neoplasias Ováricas , Humanos , Femenino , Detección Precoz del Cáncer , Dióxido de Silicio/química , Espectrometría Raman/métodos , Nanoestructuras/uso terapéutico , Nanoestructuras/química , Neoplasias Ováricas/diagnóstico , Neoplasias Ováricas/terapia , Oro/químicaRESUMEN
Ischemic stroke (IS) is the leading cause of disability and death worldwide. Owing to the aging population and unhealthy lifestyles, the incidence of cerebrovascular disease is high. Vascular risk factors include hypertension, diabetes, dyslipidemia, and obesity. Therefore, in addition to timely and effective reperfusion therapy for IS, it is crucial to actively control these risk factors to reduce the incidence and recurrence rates of IS. Evidence from human and animal studies suggests that moderate intermittent hypoxia (IH) exposure is a promising therapeutic strategy to ameliorate common vascular risk factors and comorbidities. Given the complex pathophysiological mechanisms underlying IS, effective treatment must focus on reducing injury in the acute phase and promoting repair in the recovery phase. Therefore, this review discusses the preclinical perspectives on IH conditioning as a potential treatment for neurovascular injury and highlights IH pre and postconditioning strategies for IS. Hypoxia conditioning reduces brain injury by increasing resistance to acute ischemic and hypoxic stress, exerting neuroprotective effects, and promoting post-injury repair and regeneration. However, whether IH produces beneficial effects depends not only on the hypoxic regimen but also on inter-subject differences. Therefore, we discuss the factors that may influence the effectiveness of IH treatment, including age, sex, comorbidities, and circadian rhythm, which can be used to help identify the optimal intervention population and treatment protocols for more accurate, individualized clinical translation. In conclusion, IH conditioning as a non-invasive, non-pharmacological, systemic, and multi-targeted intervention can not only reduce brain damage after stroke but can also be applied to the prevention and functional recovery of IS, providing brain protection at different stages of the disease. It represents a promising therapeutic strategy. For patients with IS and high-risk groups, IH conditioning is expected to develop as an adjunctive clinical treatment option to reduce the incidence, recurrence, disability, and mortality of IS and to reduce disease burden.
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Epileptogenesis is a dynamic process accompanied by the hippocampal neuroinflammation, aberrant neurogenesis and cognitive decline. Accordingly, anti-neuroinflammation and neuroprotective therapy may be considered in effective prophylaxis and therapeutics for epilepsy. Insulin-like growth factor-1 (IGF-1) is well acknowledged as a neuroprotective factor to promote neurogenesis and neuronal survival and synaptogenesis, wherein there has been controversy as to its implication in epilepsy. In our prior experiments, we established transgenic mice with overexpressed IGF-1 specifically in neural stem cells (NSCs). This experiment sought to investigate the implications of overexpressed IGF-1 in a PTZ-induced mouse model of chronic epilepsy. Herein we demonstrated that increasing hippocampal levels of IGF-1 sufficed to exhibit long-term anti-epileptogenesis effect on epileptic mouse model. The severity of epilepsy was assessed with onset latency, Racine score, percentage of mice with stage IV-V seizures, duration of seizures. Strikingly, IGF-1 transgenic mice showed more lessened severity than the wild-type (W-T) mice. Moreover, IGF-1 transgenic mice exhibited a lesser population of IBA-1-positive cells in CA1 of hippocampus than did W-T mice in the chronic phase, with a significant downregulation of IL-1ß, IL-6, and TNF-α in hippocampus, indicating that IGF-1 overexpression significantly mitigated inflammation in the hippocampus. In addition, Nissl staining revealed a greater population of viable neurons in CA1 of IGF-1 transgenic mice versus the W-T mice. TUNEL staining and western blotting indicated fewer apoptotic cells in transgenic mice. Moreover, we identified the evident increase in BrdU-positive and DCX-positive cell populations in dentate gyrus (DG) in IGF-1 transgenic mice, versus the W-T mice, indicative of the role of IGF-1 overexpression in promoting long-term hippocampal neurogenesis in the chronic phase of epilepsy. Collectively, our data authenticated the neuroprotective and antiinflammatory effects of IGF-1 in the chronic phase of epilepsy, which may benefit the long-term anti-epileptogenesis regimens.
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Epilepsia , Fármacos Neuroprotectores , Animales , Antiinflamatorios/farmacología , Modelos Animales de Enfermedad , Epilepsia/tratamiento farmacológico , Hipocampo , Factor I del Crecimiento Similar a la Insulina , Ratones , Ratones Transgénicos , Fármacos Neuroprotectores/farmacología , Fármacos Neuroprotectores/uso terapéutico , Pentilenotetrazol/farmacología , Convulsiones/tratamiento farmacológicoRESUMEN
Epilepsy is the most common disease of the nervous system, characterized by aberrant normal brain activity. Neuroinflammation is a prominent feature in the brain in epileptic humans and animal models of epilepsy. However, it remains elusive as to how peripheral inflammation affects epilepsy. Herein we demonstrated significantly greater seizure susceptibility and severity of epilepsy under kainic acid (KA) via intraperitoneal (i.p.) injection of lipopolysaccharide (LPS) in mouse model of epilepsy. Nissl staining was employed for assessment of the neuronal damage, immunofluorescence for staining of the microglial cells and astrocytes in the mouse brain slices, and ELISA for detection of the changes of inflammatory factors. We observed a smaller population of viable neurons in CA1 and CA3 regions, a greater population of IBA-1-positive and GFAP-positive cells, with a significant upregulation of IL-1ß and IL-6 in hippocampus of epileptic mice when treated with LPS, indicating that LPS aggravates hippocampal neuron injury in epilepsy, and induces neuroinflammation in the hippocampus. In addition, we provide an evident increase in BrdU+/DCX+ and Nestin+ cell populations in dentate gyrus (DG) in LPS-treated group, versus saline group on epileptic mouse model, which demonstrated LPS treatment enhanced hippocampal neurogenesis. In order to investigate whether interleukin-1 type 1 (IL-1R1) signaling is involved in this process, we adopted IL-1R1 globally restored mice (IL-1R1GR/GR) as an IL-1R1 reporter to visualize labeling of IL-1R1 mRNA and protein by means of RFP staining. Strikingly, the RFP immunofluorescence revealed increased IL-1R1 expression in LPS-treated group, versus saline group. Further, blockage of central IL-1R1 alleviated seizure susceptibility and severity of epilepsy. In summary, our findings suggested that LPS could enhance central inflammatory response and aggravate the susceptibility to epileptic seizure, which we postulated to be mediated by IL-1R1.
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Epilepsia , Lipopolisacáridos , Animales , Modelos Animales de Enfermedad , Epilepsia/inducido químicamente , Epilepsia/metabolismo , Hipocampo/metabolismo , Humanos , Lipopolisacáridos/metabolismo , Lipopolisacáridos/farmacología , Ratones , Ratones Endogámicos C57BL , Receptores de Interleucina-1/metabolismo , Convulsiones/metabolismoRESUMEN
Ischemic stroke is associated with increasing morbidity and has become the main cause of death and disability worldwide. Cerebral edema is a serious complication arising from ischemic stroke. It causes an increase in intracranial pressure, rapid deterioration of neurological symptoms, and formation of cerebral hernia, and is an important risk factor for adverse outcomes after stroke. To date, the detailed mechanism of cerebral edema after stroke remains unclear. This limits advances in prevention and treatment strategies as well as drug development. This review discusses the classification and pathological characteristics of cerebral edema, the possible relationship of the development of cerebral edema after ischemic stroke with aquaporin 4, the SUR1-TRPM4 channel, matrix metalloproteinase 9, microRNA, cerebral venous reflux, inflammatory reactions, and cerebral ischemia/reperfusion injury. It also summarizes research on new therapeutic drugs for post-stroke cerebral edema. Thus, this review provides a reference for further studies and for clinical treatment of cerebral edema after ischemic stroke.
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Ischemic stroke (IS) is a cerebrovascular disease with serious neurological function impairment, which may activate endoplasmic reticulum (ER) stress. However, the underlying regulatory mechanism of ER stress under IS remains unclear. miR-9-5p is enriched in the brain tissues and plays a role in the pathological process of IS. Therefore, the purpose of this study is to explore the effect of miR-9 on ER stress and underlying mechanism in IS. Here, a middle cerebral artery occlusion (MCAO) rat model was utilized to examine the alteration of brain pathology, and the expressions of miR-9 and ER stress-related proteins. Then SH-SY5Y cells with oxygen-glucose deprivation (OGD) were performed to further evaluate the functional role of miR-9 and preliminary mechanism. The results showed that miR-9 levels were decreased in the ischemic region of rats after MCAO. MCAO significantly increased the brain infract volume, reduced Nissl bodies and cell apoptosis, and increased ER stress-related proteins (ERMP1, GRP78, p-PERK, p-eIF2α and CHOP). Furthermore, overexpression of miR-9 by miR-9 mimics increased cell viability, inhibited LDH activity and cell apoptosis, and inactivated ER stress in OGD-neurons. Luciferase activity results showed that miR-9 negatively regulated ERMP1 expression by directly targeting ERMP1 3' UTR. Subsequently, we found that ERMP1 overexpression reversed the inhibition of miR-9 on GRP78-PERK-CHOP pathway in OGD neurons. In summary, our results suggest that the attenuation of miR-9 on ischemic injury may be involved in targeting ERMP1-mediated ER stress, which provides an available target for IS treatment.
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Infarto Encefálico/metabolismo , Estrés del Retículo Endoplásmico , Proteínas de la Membrana/biosíntesis , Metaloproteasas/biosíntesis , MicroARNs/metabolismo , Accidente Cerebrovascular/metabolismo , Animales , Infarto Encefálico/patología , Línea Celular Tumoral , Modelos Animales de Enfermedad , Masculino , Ratas , Ratas Sprague-Dawley , Accidente Cerebrovascular/patologíaRESUMEN
Ovariectomy disrupts estrogen production and homeostasis. However, whether exercise training (ET) could counteract the ovariectomy-induced effect on muscular autophagy has remained elusive. This study examined muscular autophagy in ovariectomized (OVX) rats following 8 weeks of swimming ET. Here, 40 6-month-old female Sprague-Dawley rats were randomly divided into five groups: sham-operated control (Sham), OVX control (OVX), OVX with 60-min ET (OVX-60ET), 90-min ET (OVX-90ET), and 120-min ET (OVX-120ET) for 6 days/week. According to the results of Western blotting, the expression levels of autophagy-related proteins in the OVX gastrocnemius muscle, including mammalian target of rapamycin, uncoordinated 51-like kinase 1, Beclin-1, autophagy-related gene (Atg-7), and microtubule-associated protein light chains 3 were significantly decreased (all P < 0.05), while there was an elevation on the p62 level. ET appreciably mitigated the OVX-induced negative effects on muscle quality and the autophagy pathway, which seemed to be dependent on ET volume. The most optimal outcomes were observed in the OVX-90ET group. The OVX-120 group had an adversely augmented catabolic process associated with gastrocnemius muscle atrophy. In conclusion, the expression levels of autophagy proteins are decreased in OVX rats, which can be appreciably mitigated following 8 weeks of swimming ET.
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Although immunosuppressive therapies have made organ transplantation a common medical procedure worldwide, chronic toxicity is a major issue of long-term treatment. One method to improve such therapies is the application of immunomodulatory agents from parasites, such as Hypoderma lineatum (Diptera: Oestridae). Hypodermin C (HC) is an enzyme secreted by H. lineatum larvae, and our previous study showed that recombinant HC could degrade guinea pig C3 and inhibit the complement pathway in vitro, suggesting potential activity for inhibiting transplant rejection. However, such properties have not been fully demonstrated in vivo. In this study, we investigated the impact of HC on a fully MHC-mismatched, life-sustaining, murine model of kidney allograft rejection using B6 donors and BABL/c (HC transgenic or wild-type) recipients. Kidney grafts were analyzed by histology, immunohistochemistry and western blotting. The results suggested that HC could effectively inhibit kidney allograft rejection. These findings suggest HC is a promising strategy to improve the survival of human implants.
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Rechazo de Injerto/tratamiento farmacológico , Supervivencia de Injerto/efectos de los fármacos , Inmunosupresores/uso terapéutico , Trasplante de Riñón , Serina Endopeptidasas/uso terapéutico , Aloinjertos/inmunología , Animales , Enfermedad Crónica , Dípteros/inmunología , Modelos Animales de Enfermedad , Antígenos de Histocompatibilidad/inmunología , Humanos , Inmunomodulación , Terapia de Inmunosupresión , Larva , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Transgénicos , Serina Endopeptidasas/genéticaRESUMEN
Cyclodextrin (ß-CD)-functionalized reduced graphene oxide was successfully synthesized by a simple hydrothermal method, followed by conjugating with polyethylene glycol (PEG) and folic acid (FA). Microscopic and spectroscopic techniques were used to characterize the nanocomposites. Photothermal experiments showed that ß-CD-functionalized reduced graphene oxide exhibited higher photothermal conversion efficiency in the near infrared region than reduced graphene oxide functionalized with other molecules under the same conditions. Cytotoxicity experiments indicated that rGO@CD@PEG@FA possessed good biocompatibility even at high concentration. When doxorubicin (DOX) was loaded on the rGO@CD@PEG@FA nanocomposite, it showed the stimulative effect of heat, pH response, and sustained drug release. Cytotoxicity experiments also confirmed the targeted effect and high efficiency of the combined therapy. The findings of the present study provide an ideal drug delivery system for malignant cancer therapy due to the advanced synergistic chemo-photothermal targeted therapy and good drug release properties.
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Diabetic retinopathy is the most universal and severe complication of diabetes mellitus. The primary aim of the present study was to determine whether tanshinone IIA (TSA) has an inhibitory effect on the proliferation, migration and vascularization of human retinal endothelial cells (HREC) under high glucose (HG) conditions and the associated underlying mechanism. It was demonstrated that TSA exhibited a significant inhibitory effect on the proliferation, migration and vascularization of HRECs in a dosedependent manner, under conditions of high glucose (25 mM) medium. However, there was no distinct inhibitory effect on HREC proliferation, migration and vascularization under normal glucose (NG, 5.5 mM glucose) conditions. Reverse transcriptionquantitative polymerase chain reaction, western blotting and immunofluorescence experiments were conducted to evaluate the effects of TSA on the expression levels of vascular endothelial growth factor (VEGF) and intercellular adhesion molecule (ICAM)1. It was demonstrated that TSA significantly downregulated the expression levels of VEGF and ICAM1 in a dosedependent manner under HG conditions. Overall, the results of the present study indicate that TSAmediated inhibition of proliferation, migration and vascularization in HRECs may be associated with its ability to affect the expression levels of VEGF and ICAM1.
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Abietanos/farmacología , Movimiento Celular/efectos de los fármacos , Células Endoteliales/citología , Glucosa/toxicidad , Neovascularización Fisiológica/efectos de los fármacos , Retina/citología , Línea Celular , Proliferación Celular/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Células Endoteliales/efectos de los fármacos , Células Endoteliales/metabolismo , Humanos , Molécula 1 de Adhesión Intercelular/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
The neurotrophic factor insulin-like growth factor (IGF)-1 promotes neurogenesis in the mammalian brain and provides protection against brain injury. However, studies regarding the effects of IGF-1 on cognitive function in aged mice remain limited. We investigated the effects of overexpression of IGF-1 specifically in neural stem cells of the hippocampal dentate gyrus on the recognitive function in 18-month-old transgenic mice. Immunohistocytochemistry and Nissl staining revealed the increased population of BrdU-positive cells as well as the upregulated expression of Nestin and neuronal nuclei (NeuN), respective markers for neural progenitors and neurons, in the hippocampus of the aged IGF-1 transgenic mice versus the wild-type, suggesting that IGF-1 overexpression promotes neurogenesis. In addition, the IGF-1 receptor (IGF-1R), the phosphorylation of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) and extracellular signal-regulated kinase (ERK) were enhanced in the transgenic mice than in the wild-type. Transgenic mice also showed superior performance in the Morris water maze and step-down memory tests to their wild-type counterparts. Moreover, the learning and memory abilities of transgenic mice were significantly undermined with the blockage of CaMKII and ERK signaling pathway. Accordingly, our findings indicated that IGF-1 may mitigate the aged-associated cognitive decline via promoting neurogenesis in the hippocampus and activating CaMKII and ERK signaling by binding with IGF-1R.
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Proteína Quinasa Tipo 2 Dependiente de Calcio Calmodulina/metabolismo , Cognición/fisiología , Factor I del Crecimiento Similar a la Insulina/biosíntesis , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Animales , Giro Dentado/metabolismo , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Expresión Génica , Hipocampo/metabolismo , Aprendizaje por Laberinto , Memoria/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Células-Madre Neurales/metabolismo , Neurogénesis , Neuronas/metabolismo , Fosforilación , Receptor IGF Tipo 1/metabolismo , Transducción de Señal , Regulación hacia ArribaRESUMEN
Growth factors like insulin-like growth factor 1 (IGF-1) is reported to mediate neurogenesis in the subgranular zone (SGZ) and the subventricular zone (SVZ) of the adult mammalian brain, but its regulatory mechanism remains unclear. We generated transgenic mice overexpressing IGF-1 specifically in neural stem cells (NSCs) and assessed the effect of IGF-1 on neurogenesis in adult mice NSCs. Overexpression of IGF-1 could stimulate the expression of phospho-Akt and phospho-ERK1/2 while inducing proliferation and differentiation of NSCs in the SGZ and SVZ. The MEK/ERK inhibitor U0126 could inhibit ERK1/2 phosphorylation, further inhibiting the proliferation of NSCs in the SGZ and SVZ but had no effect on the phosphorylation of Akt. By contrast, The PI3K/Akt inhibitor LY294002 inhibited phosphorylation of Akt and differentiation of NSCs in the SGZ and SVZ, resulting in no change in the proliferation of NSCs and ERK1/2 phosphorylation. These results demonstrate that IGF-1 upregulates the proliferation of NSCs by triggering MEK/ERK pathway signaling in the adult mice SGZ and SVZ. Meanwhile, IGF-1 also induces differentiation of NSCs via the PI3K/Akt pathway in adult mice. However, we found no evidence of crosstalk between the PI3K/Akt and MEK/ERK pathways in adult mice NSCs. Our work provides new experimental evidence of the involvement of the PI3K/Akt and MEK/ERK pathways in the proliferation and differentiation of the NSCs of adult mice.
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Factor I del Crecimiento Similar a la Insulina/biosíntesis , Células-Madre Neurales/fisiología , Neurogénesis/fisiología , Células Madre Adultas/fisiología , Animales , Proliferación Celular/fisiología , Células Cultivadas , Ratones , Ratones Endogámicos C57BL , Ratones TransgénicosRESUMEN
Immunosuppressive agents play an important role in the success of organ transplantation, however the chronic toxicity of these agents is a major issue over the long-term. Hypodermin A (HA) is an enzyme secreted by the larvae of Hypoderma lineatum (Diptera: Oestridae), and has been implicated in immunosuppression in cattle. Malassagne et al. have demonstrated that HA can degrade the C3 protein, and could be used to prevent hyperacute xenogeneic rejection. We found that overexpression of HA in RAW264.7 cells induced significant secretion of prostaglandin E2 (PGE2), which mediates a variety of innate and adaptive immune responses through four E-type prostanoid (EP) receptor subtypes (EP1-4). PGE2 is useful in the management of allogeneic acute rejection. In addition, we found that induction of PGE2 expression downregulates the expression of interferon (IFN)-γ and interleukin (IL)-2, and promotes the secretion of IL-10 in vitro through the EP4 receptor. It was previously shown that activation of IL-2 and IFN-γ is involved in allograft acute rejection. IL-10 is known to prevent inflammation, and can improve allograft survival rates. We concluded that besides preventing hyperacute xenogeneic rejection, HA might also be a potential therapeutic candidate for ameliorating acute rejection during allotransplantation.
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Rechazo de Injerto/terapia , Macrófagos/fisiología , Serina Endopeptidasas/metabolismo , Enfermedad Aguda , Animales , Bovinos , Línea Celular , Citocinas/metabolismo , Dinoprostona/metabolismo , Dípteros/inmunología , Rechazo de Injerto/inmunología , Humanos , Terapia de Inmunosupresión , Ratones , Subtipo EP4 de Receptores de Prostaglandina E/metabolismo , Serina Endopeptidasas/genética , Transgenes/genéticaRESUMEN
Pigment epithelium-derived factor (PEDF) has many biological activities. But it's not known whether PEDF and its functional peptides could protect against hypoxia-induced cell death and the mechanisms are still unclear. We used cultured H9c2 cells and primary cardiomyocytes to show that apoptosis and necroptosis were significantly increased after hypoxia. Both PEDF and its fuctional peptides 44mer reduced apoptosis and necroptosis rates and inhibited the expression of cleaved caspase 3 and receptor-interacting protein 3 (RIP3). Furthermore, PEDF and 44mer could up-regulate super oxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) levels, promote clearing of reactive oxygen species (ROS) and malondialdehyde (MDA). While, 34mer, another functional peptides had no effect on cell apoptosis and necroptosis. Hereby this is the first evidence that PEDF and its functional peptide 44mer protect cultured H9c2 cells and primary cardiomyocytes against apoptosis and necroptosis under hypoxic condition via the anti-oxidative mechanism.
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Antioxidantes/metabolismo , Apoptosis/fisiología , Proteínas del Ojo/metabolismo , Hipoxia/metabolismo , Miocitos Cardíacos/metabolismo , Necrosis/metabolismo , Factores de Crecimiento Nervioso/metabolismo , Péptidos/metabolismo , Serpinas/metabolismo , Animales , Caspasa 3/metabolismo , Células Cultivadas , Glutatión Peroxidasa/metabolismo , Estrés Oxidativo/fisiología , Ratas , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno/metabolismo , Proteína Serina-Treonina Quinasas de Interacción con Receptores/metabolismo , Superóxido Dismutasa/metabolismoRESUMEN
Heme oxygenase (HO) represents an intrinsic antiinflammatory system based on its ability to inhibit expression of proinflammatory cytokines. The constitutive isoform heme oxygenase-2 (HO-2) has high expression and activity in cerebral microvascular endothelial cells (CMVEC). This study was undertaken to evaluate the role of HO-2 in regulation of TLR4/MyD88-dependent signaling and to study the effect of HO-2 on the expression and secretion of the proinflammatory cytokines tumor necrosis factor α (TNF-α) and Interleukin-6 (IL6) in CMVEC. HO-2 short hairpin RNA (shRNA) and HO-2 overexpression plasmids were used to observe the effect of HO-2 on proinflammatory cytokines in CMVEC in vitro, and the results showed that the messenger RNA (mRNA) and protein levels of TNF-α and IL6 were increased and decreased, respectively, compared with control groups. LPS-stimulated TNF-α and IL6 mRNA and protein were also reduced in CMVEC treated with an inhibitor of TLR4 signaling, CLI-095, or HO-2 overexpression. CLI-095 and HO-2 overexpression both reduced TLR4 expression in CMVEC, and HO-2 shRNA blocked these effects of CLI-095. CLI-095 and HO-2 overexpression potently suppressed TLR4/MyD88-dependent proinflammatory cytokine expression in CMVEC. These results suggest that HO-2 plays an important role in protecting CMVEC against cytokine-mediated inflammation.
Asunto(s)
Células Endoteliales/metabolismo , Endotelio Vascular/metabolismo , Hemo Oxigenasa (Desciclizante)/metabolismo , Interleucina-6/metabolismo , Transducción de Señal/fisiología , Receptor Toll-Like 4/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Animales , Células Cultivadas , Corteza Cerebral/citología , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/metabolismo , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Hemo Oxigenasa (Desciclizante)/genética , Interleucina-6/genética , Lipopolisacáridos/farmacología , Ratones , ARN Interferente Pequeño , Transducción de Señal/efectos de los fármacos , Sulfonamidas/farmacología , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
Hypodermin A (HA), a serine protease secreted by first-instar larvae of Hypoderma lineatum (Diptera: Oestridae) is associated with inflammatory and the specific immune responses in cattle hosts. In the present study, the cDNA sequence of HA was synthesized, and found to have fifteen amino acids which differed from the sequence available in GenBank. We then examined the association between recombinant HA and guinea-pig complement component 3 (C3) through a co-immunoprecipitation assay. Cos7 cells stably expressing HA were generated, and were found to be more resistant to lysis by guinea-pig C3 than the controls. HA was also able to degrade the C6 and C5b-9 of guinea-pig C3. The presumed DNA binding site of HA with guinea-pig C3 was detected by an electrophoretic mobility shift assay (EMSA). In contrast, after stable transfection, mHA was unable to reduce the amount of C3 or to inhibit its cytotoxicity, while HA could degrade guinea-pig C3 and inhibit the complement pathway. The findings suggest that recombinant HA could serve as an immunosuppressive agent against organ rejection after xenotransplantation.
Asunto(s)
Proteínas del Sistema Complemento/metabolismo , Serina Endopeptidasas/farmacología , Secuencia de Aminoácidos , Animales , Células COS , Chlorocebus aethiops , ADN Complementario/genética , Datos de Secuencia MolecularRESUMEN
Studies have shown that estrogen has neuroprotective effects on the nigrostriatal system. The present study established a Parkinson's disease model in C57BL/6 mice by intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrapyridine. The mice were subjected to 17ß estradiol injection into the lateral ventricle. Immunofluorescence double staining showed that estrogen increased tyrosine hydroxylase and calbindin-D28K expression and co-expression in dopaminergic neurons of midbrain substantia nigra pars compacta of model mice. Behavior experiments showed that estrogen improved swimming and hanging behaviors in this mouse model of Parkinson's disease.