RESUMO
Currently, there are no effective therapeutic targets for the treatment of chronic cerebral hypoperfusion(CCH)-induced cerebral ischemic injury. Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are discovered as the inducers of neurogenesis and angiogenesis. We previously made a nanofiber membrane (NFM), maintaining a long-term release of VEGF and bFGF up to 35 days, which might make VEGF and bFGF NFM as the potential protective agents against cerebral ischemic insult. In this study, the effects of VEGF and bFGF delivered by NFM into brain were investigated as well as their underlying mechanismsin a rat model of CCH. VEGF + bFGF NFM application increased the expressions of tight junction proteins, maintained BBB integrity, and alleviated vasogenic cerebral edema. Furthermore, VEGF + bFGF NFM sticking enhanced angiogenesis and elevated CBF. Besides, VEGF + bFGF NFM treatment inhibited neuronal apoptosis and decreased neuronal loss. Moreover, roofing of VEGF + bFGF NFM attenuated microglial activation and blocked the launch of NLRP3/caspase-1/IL-1ß pathway. In addition, VEGF + bFGF NFM administration prevented disruption to the pre/postsynaptic membranes and loss of myelin sheath, relieving synaptic injury and demyelination. Oligodendrogenesis, neurogenesis and PI3K/AKT/mTOR pathway were involved in the treatment of VEGF + bFGF NFM against CCH-induced neuronal injury and hypomyelination. These findings supported that VEGF + bFGF NFM application constitutes a neuroprotective strategy for the treatment of CCH, which may be worth further clinical translational research as a novel neuroprotective approach, benifiting indirect surgical revascularization.
Assuntos
Lesões Encefálicas , Isquemia Encefálica , Nanofibras , Ratos , Animais , Fator A de Crescimento do Endotélio Vascular/uso terapêutico , Fator A de Crescimento do Endotélio Vascular/metabolismo , Fator 2 de Crescimento de Fibroblastos/uso terapêutico , Fosfatidilinositol 3-Quinases , Nanofibras/uso terapêutico , Fatores de Crescimento do Endotélio Vascular , Isquemia Encefálica/metabolismo , IsquemiaRESUMO
AIMS: Clear roles and mechanisms in explaining gut microbial dysbiosis and microbial metabolites short-chain fatty acids (SCFAs) alterations in chronic cerebral ischemic pathogenesis have yet to be explored. In this study, we investigated chronic cerebral hypoperfusion (CCH)-induced gut microbiota and metabolic profiles of SCFAs as well as the effects and mechanisms of fecal microbiota transplantation (FMT) and SCFAs treatment on CCH-induced hippocampal neuronal injury. METHODS: Bilateral common carotid artery occlusion (BCCAo) was used to establish the CCH model. Gut microbiota and SCFAs profiles in feces and hippocampus were evaluated by 16S ribosomal RNA sequencing and gas chromatography-mass spectrometry. RNA sequencing analysis was performed in hippocampal tissues. The potential molecular pathways and differential genes were verified through western blot, immunoprecipitation, immunofluorescence, and ELISA. Cognitive function was assessed via the Morris water maze test. Ultrastructures of mitochondria and synapses were tested through a transmission electron microscope. RESULTS: Chronic cerebral hypoperfusion induced decreased fecal acetic and propionic acid and reduced hippocampal acetic acid, which were reversed after FMT and SCFAs administration by changing fecal microbial community structure and compositions. Furthermore, in the hippocampus, FMT and SCFAs replenishment exerted anti-neuroinflammatory effects through inhibiting microglial and astrocytic activation as well as switching microglial phenotype from M1 toward M2. Moreover, FMT and SCFAs treatment alleviated neuronal loss and microglia-mediated synaptic loss and maintained the normal process of synaptic vesicle fusion and release, resulting in the improvement of synaptic plasticity. In addition, FMT and SCFAs supplement prevented oxidative phosphorylation dysfunction via mitochondrial metabolic reprogramming. The above effects of FMT and SCFAs treatment led to the inhibition of CCH-induced cognitive impairment. CONCLUSION: Our findings highlight FMT and SCFAs replenishment would be the feasible gut microbiota-based strategy to mitigate chronic cerebral ischemia-induced neuronal injury.
Assuntos
Isquemia Encefálica , Disfunção Cognitiva , Ratos , Animais , Transplante de Microbiota Fecal/métodos , Fezes/química , Ácidos Graxos Voláteis/análise , Isquemia Encefálica/terapia , Disfunção Cognitiva/etiologia , Disfunção Cognitiva/terapiaRESUMO
We investigated the potential effects and mechanisms of vascular endothelial growth factor (VEGF)-nanofiber membranes (NFMs) treatment in a rat model of chronic cerebral hypoperfusion (CCH). VEGF-NFMs treatment promoted angiogenesis in surgical temporal cortex and hippocampus, alleviating decreased CBF in these two cerebral regions. VEGF-NFMs application improved reduced NAA/Cr ratio, preventing neuronal loss. VEGF-NFMs sticking decreased the number of TUNEL-positive cells in surgical temporal cortex, ameliorated impaired synaptic plasticity, and inhibited the release of pro-inflammatory cytokines and the activation of microglia and astrocytes in surgical temporal cortex and hippocampus. Furthermore, BDNF-TrkB/PI3K/AKT, BDNF-TrkB/ERK and HIF-1a/VEGF/ERK pathways were involved in the treatment of VEGF-NFMs against CCH-induced neuronal injury. These results showed the neuroprotective effects of VEGF-NFMs sticking may initiate from neurovascular repairing followed by inhibition of neuronal apoptosis and neuronal and synaptic damage, eventually leading to the suppression of cognitive dysfunction, which provided theoretical foundation for further clinical transformation of VEGF-NFMs.
Assuntos
Isquemia Encefálica , Disfunção Cognitiva , Nanofibras , Ratos , Animais , Fator A de Crescimento do Endotélio Vascular/metabolismo , Fosfatidilinositol 3-Quinases , Fator Neurotrófico Derivado do Encéfalo , Isquemia Encefálica/tratamento farmacológicoRESUMO
BACKGROUND: Little is known about the association between gut microbiota and intestinal injury under a state of chronic cerebral hypoperfusion (CCH). Here, the effects of gut microbiota and short-chain fatty acids (SCFAs), as important metabolic products, on intestinal function and potential mechanisms after CCH were investigated. METHODS: Rats were subjected to bilateral common carotid artery occlusion (BCCAo) to induce CCH. The gut microbiota and metabolites of SCFAs were assessed by 16S rRNA sequencing and targeted metabolomics, respectively. Transcriptomic analysis of colon tissues was also conducted. Subsequently, potential molecular pathways and differentially expressed genes were verified by western blot, immunoprecipitation, and immunofluorescence analyses. Furthermore, the integrity of the colonic barrier was evaluated by hematoxylin and eosin and mucin 2 staining and expression levels of tight junction proteins. Besides, colonic inflammation was further assessed by flow cytometry and expression levels of inflammatory cytokines. In addition, colonic mitochondrial dysfunction was analyzed via membrane potential, reactive oxygen species, electron transport chain (ETC) activities, adenosine triphosphate content, and mitochondrial ultrastructure. RESULTS: CCH modified gut microbial composition and microbial metabolism of SCFAs, which may be associated with inhibition of mitochondrial ETC activities and oxidative phosphorylation, leading to dysregulation of mitochondrial energy metabolism. Furthermore, CCH induced differentiation of pathogenic Th17 cells, promoted the formation of complexes of interferon regulatory factor 4 and signal transducer and activator of transcription 3 (STAT3), and increased the phosphorylation of STAT3. This was associated with an impairment of colonic barrier function and chronic colonic inflammation. In contrast, FMT and SCFA replenishment ameliorated CCH-induced gut microbial dysbiosis by increasing the intestinal content of Ruminococcus_sp_N15_MGS_57 and modulating microbial metabolism of SCFAs by increasing acetic acid contents associated with an improvment of the balance between Tregs and Th17 cells, mitochondrial ETC activities, and oxidative phosphorylation to prevent colonic inflammation and dysregulation of mitochondrial energy metabolism. CONCLUSION: These findings indicate that FMT and SCFA replenishment present a promising therapeutic strategy against colonic dysfunction under a state of chronic cerebral ischemia.
Assuntos
Isquemia Encefálica , Microbioma Gastrointestinal , Ratos , Animais , Microbioma Gastrointestinal/fisiologia , Transplante de Microbiota Fecal , Células Th17/metabolismo , RNA Ribossômico 16S/metabolismo , Colo/química , Colo/metabolismo , Ácidos Graxos Voláteis/metabolismo , Inflamação/metabolismo , Isquemia Encefálica/metabolismo , Metabolismo Energético , Mitocôndrias/metabolismo , Diferenciação CelularRESUMO
BACKGROUND: Chronic cerebral hypoperfusion (CCH) is a major risk factor for vascular dementia (VaD). There are currently no broadly effective prevention or treatment strategies for VaD, but recent studies have reported promising results following vascular bypass surgery and pharmacomodulation of the brain endocannabinoid system (ECS). In this study, early effects of encephalomyosynangiosis (EMS) bypass surgery and augmented endocannabinoid signaling on CCH-induced cognitive dysfunction and neuronal damage were investigated. METHODS: An animal model of VaD was established by bilateral common carotid artery occlusion (BCCAO). Cannabinoid signaling was upregulated by treatment with the fatty acid amide hydrolase inhibitor URB597 (URB). Spatial learning and memory, cerebral blood flow (CBF), revascularization, brain-derived neurotrophic factor (BDNF)-tropomyosin receptor kinase B (TrkB) signaling, and apoptosis were compared among Sham, BCCAO, BCCAO + EMS, BCCAO + URB, and BCCAO + URB + EMS groups. Spatial learning and memory were evaluated using the Morris water maze (MWM). The CBF in cortex and hippocampus was evaluated by 3-dimensional arterial spin labeling. The neovascularization was visualized by CD34 immunofluorescence staining, and BDNF-TrkB signaling protein expression levels were assessed by Western blotting. RESULTS: Treatment with URB597 but not EMS alone reversed the spatial learning and memory deficits induced by BCCAO. Neovascularization was enhanced after EMS surgery but not by URB597. Alternatively, there were no significant differences in CBF among treatment groups. Expression levels of BDNF and TrkB were significantly reduced by CCH compared to Sham treatment, and downregulation of both proteins was reversed by URB597 treatment but not EMS. BCCAO enhanced neuronal apoptosis, which was also reversed by URB597. CONCLUSIONS: Augmentation of endogenous cannabinoid signaling but not EMS protects against CCH-induced neurodegeneration and preserves spatial learning and memory, possibly by activating BDNF-TrkB signaling.
RESUMO
OBJECTIVE: To investigate the effect of electroacupuncture (EA) at "Baihui"(GV20), "Fengfu"(GV16) and bilateral "Shenshu"(BL23) on learning-memory ability, apoptosis in the hippocampus and expression of Aß, Caspase 3, Bax and Bcl-2 proteins in the hippocampus and cerebral cortex in immature mice with Alzheimer's disease (AD), so as to explore its mechanism underlying improvement of AD. METHODS: Forty APP/PS1 transgenic male young mice were equally randomized into model and EA groups and 20 C57BL/6J male young mice were used as the normal control. EA (10 Hz, about 2 mA) was applied to GV20-BL23 and GV16-BL23 for 20 min, once daily, 6 days a week for 16 weeks. The Morris water maze swimming test was used to evaluate the animals' learning-memory ability. Congo red staining and immunohistochemical staining were used to detect senile plaques in the hippocampus (dentate gyrus) and cerebral cortex tissues. Terminal deoxynucleotidyl transferase-mediated dUTP Nick-end Labeling (TUNEL) was used to detect the cellular apoptosis of hippocampus. The expression levels of apoptosis related factors Caspase 3, Bax and Bcl-2 were detected by Western blot. RESULTS: After modeling, the escape latency of place navigation test of Morris water maze swimming tasks was significantly increased (P<0.05), while the number of platform crossing and residence time in the platform quadrant of spatial exploration test were significantly decreased in the model group in contrast to the normal control group (P<0.05). The number of apoptotic cells in the hippocampus and expression levels of Aß, Caspase 3 and Bax proteins in the hippocampus and cerebral cortex were significantly up-regulated in the model group relevant to the normal control group (P<0.05). Following EA intervention, the escape latency of place navigation test of Morris water maze swimming tasks was significantly decreased (P<0.05), while the number of platform crossing and residence time in the platform quadrant of spatial exploration test were significantly increased in the EA group in contrast to the model group (P<0.05). The hippocampal apoptotic cells, the expression of Aß, Caspase 3 and Bax proteins in hippocampus and cerebral cortex were evidently down-regulated in the EA group in contrast to the model group (P<0.05). Whereas the ratio of Bcl-2/Bax was significantly decreased in the model group relevant to the normal control group (P<0.05) and considerably increased in the EA group in contrast to the model group (P<0.05). No significant changes were found in the expression levels of Bcl-2 after modeling and after EA intervention (P>0.05). CONCLUSION: EA of GV20, GV16 and BL23 can effectively improve the learning-memory ability in AD mice, which may be related to its function in inhibiting neuronal apoptosis in the hippocampus and down-regulating the expression levels of Aß, Caspase 3 and Bax proteins in both hippocampus and cerebral cortex.
Assuntos
Doença de Alzheimer , Eletroacupuntura , Animais , Apoptose , Córtex Cerebral , Hipocampo , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
To explore the potential effects of andrographolide on chronic cerebral hypoperfusion (CCH)-induced neuronal damage as well as the underlying mechanisms. Rat CCH model was established by 2-vessel occlusion (2VO). The CCH rats received andrographolide treatment for 4 weeks. The neuron loss was detected by using neuronal nuclei (NeuN) immunofluorescent staining. The expression levels of phospho-phosphatase and tensin homolog deleted on chromosome ten (p-PTEN), protein kinase B (AKT), p-AKT, and cysteinyl aspartate specific proteinase-3 (Caspase-3) proteins were accessed by Western blotting. Moreover, the neuronal apoptosis of hippocampus tissues was detected via terminal deoxynucleotidyl transferase- mediated dUTP nick end labeling (TUNEL) staining. CCH reduced the number of NeuN-positive cells, while the number was significant increased after andrographolide treatment. CCH increased the proteins expression level of p-PTEN, Caspase-3, and decreased the p-AKT, which were reversed by andrographolide treatment. Furthermore, andrographolide treatment also down-regulated CCH-induced TUNEL-apoptosis rate. Our results suggest that the PTEN/AKT pathway may be modulated by andrographolide and the damaging effects of CCH on hippocampus may be ameliorated by andrographolide treatment. Andrographolide may act as a potential therapeutic approach for chronic ischemic insults.
Assuntos
Isquemia Encefálica/tratamento farmacológico , Isquemia Encefálica/patologia , Diterpenos/uso terapêutico , Hipocampo/patologia , Neurônios/patologia , Fármacos Neuroprotetores/uso terapêutico , Proteína Oncogênica v-akt/efeitos dos fármacos , PTEN Fosfo-Hidrolase/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Animais , Antígenos Nucleares/metabolismo , Caspase 3/efeitos dos fármacos , Doença Crônica , Hipocampo/enzimologia , Marcação In Situ das Extremidades Cortadas , Masculino , Proteínas do Tecido Nervoso/metabolismo , Neurônios/enzimologia , Ratos , Ratos Sprague-DawleyRESUMO
The purpose of this study was to explore the effects of calcitonin gene-related peptide (CGRP) on the long-term depression (LTD) of hippocampus in mice. Sixty C57BL/6J mice (30 days old) were randomly divided into control group, three CGRP (50, 100, and 200 nmol/L) groups, CGRP + CGRP8-37 group and CGRP + APV group (10 mice for each group). The effects of exogenous application of different concentrations of CGRP on synaptic plasticity and LTD in hippocampus of mice were detected by in vitro recording of local field potential. The results showed that higher doses (100 and 200 nmol/L) of CGRP significantly enhanced the induction of LTD in the hippocampus. Moreover, CGRP increased the magnitude of N-methyl-D-aspartate (NMDA) receptor-mediated excitatory postsynaptic currents. The above-mentioned effects of CGRP were blocked by either CGRP selective antagonist CGRP8-37 or NMDA receptor antagonist APV. These results suggest that CGRP can dose-dependently enhance the induction of LTD in hippocampus of mice, and the underlying mechanism involves the mediation of NMDA receptor function.
Assuntos
Peptídeo Relacionado com Gene de Calcitonina , Hipocampo , Animais , Peptídeo Relacionado com Gene de Calcitonina/farmacologia , Hipocampo/efeitos dos fármacos , Depressão Sináptica de Longo Prazo/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Distribuição AleatóriaRESUMO
BACKGROUND: Previous studies reported that URB597 (URB) had therapeutic potential for treating chronic cerebral hypoperfusion (CCH)-induced neuroinflammation and autophagy dysfunction. However, the interaction mechanisms underlying the CCH-induced abnormal excessive autophagy and neuroinflammation remain unknown. In this study, we investigated the roles of impaired autophagy in nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing (NLRP) 3 inflammasome activation in the rat hippocampus and the underlying mechanisms under the condition of induced CCH as well as the effect of URB treatment. METHODS: The CCH rat model was established by bilateral common carotid artery occlusion (BCCAo), and rats were randomly divided into 11 groups as follows: (1) sham-operated, (2) BCCAo; (3) BCCAo+autophagy inhibitor 3-methyladenine (3-MA), (4) BCCAo+lysosome inhibitor chloroquine (CQ), (5) BCCAo+microglial activation inhibitor minocycline, (6) BCCAo+ROS scavenger N-acetylcysteine (NAC), (7) BCCAo+URB, (8) BCCAo+URB+3-MA, (9) BCCAo+URB+CQ, (10) BCCAo+URB+minocycline, (11) BCCAo+URB+NAC. The cell localizations of LC3, p62, LAMP1, TOM20 and NLRP3 were assessed by immunofluorescence staining. The levels of autophagy-related proteins (LC3, p62, LAMP1, BNIP3 and parkin), NLRP3 inflammasome-related proteins (NLRP3, CASP1 and IL-1ß), microglial marker (OX-42) and proinflammatory cytokines (iNOS and COX-2) were evaluated by western blotting, and proinflammatory cytokines (IL-1ß and TNF-a) were determined by ELISA. Reactive oxygen species (ROS) were assessed by dihydroethidium staining. The mitochondrial ultrastructural changes were examined by electron microscopy. RESULTS: CCH induced microglial overactivation and ROS accumulation, promoting the activation of the NLRP3 inflammasome and the release of IL-1ß. Blocked autophagy and mitophagy flux enhanced the activation of the NLRP3-CASP1 inflammasome pathway. However, URB alleviated impaired autophagy and mitophagy by decreasing mitochondrial ROS and microglial overactivation as well as restoring lysosomal function, which would further inhibit the activation of the NLRP3-CASP1 inflammasome pathway. CONCLUSION: These findings extended previous studies indicating the function of URB in the mitigation of chronic ischemic injury of the brain.
Assuntos
Autofagia/fisiologia , Benzamidas/uso terapêutico , Isquemia Encefálica/metabolismo , Carbamatos/uso terapêutico , Modelos Animais de Doenças , Inflamassomos/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Animais , Autofagia/efeitos dos fármacos , Benzamidas/farmacologia , Isquemia Encefálica/prevenção & controle , Carbamatos/farmacologia , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Inflamassomos/antagonistas & inibidores , Masculino , Proteína 3 que Contém Domínio de Pirina da Família NLR/antagonistas & inibidores , Fármacos Neuroprotetores/farmacologia , Fármacos Neuroprotetores/uso terapêutico , Distribuição Aleatória , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVE: To observe the influence of different courses of electroacupuncture (EA) intervention on recognition memory and the proliferation and differentiation of hippocampal neural stem cells in mice with radiation-induced brain injury, so as to explore its mechanisms underlying improving radiation-induced brain injury. METHODS: Se-venty 30-day old C57BL/6J mice were randomly divided into control, model and EA groups, and the latter two groups were further divided into 1 week (W), 2 W and 3 W subgroups (n=10 in the control group and each subgroup). The ra-diation-induced brain injury model was established by radiating the mouse' left head at a dose of 8 Gy for 10 min by using a radiation linear accelerator. EA (1.5 V, 2 Hz/10 Hz) was applied to "Baihui" (GV20), "Fengfu" (GV14) and bilateral "Shenshu" (BL23) for 30 min, once daily for 1, 2 and 3 weeks, respectively. The learning-cognition memory ability was detected by using novel object recognition test in an open test box to record the time for exploring a novel object (TN) and a familiar object and to calculate the recognition index (RI). The neural stem cells' proliferation and differentiation in the hippocampus tissues were evaluated by counting the number of bromodeoxyuridine (BrdU)-labeled cells, neuronal nuclei (NeuN)/BrdU-positive cells and BrdU/glia fibrillary acidic protein (GFAP)-positive cells under microscope after immunofluorescence stain. RESULTS: After modeling, the TN at 90 min and 24 h and RI of the model subgroup 3 W at 90 min and RI of the model subgroup 1, 2 and 3 W at 24 h were significantly decreased in comparison with those of the control group (P<0.01, P<0.05). Moreover, the number of BrdU-positive cells in the model subgroup 1 W and 2 W, the BrdU/NeuN double-labeled cells in the 3 model subgroups and BrdU/GFAP double-labeled cells in the model subgroup 1 W and 3 W were significantly decreased (P<0.01, P<0.05). Following EA interventions, the TN in the 3 EA subgroups at both 90 min and 24 h, and RI of EA subgroup 3 W at 90 min and EA subgroup 2 W and 3 W at 24 h were considerably increased compared with those of the corresponding 3 model subgroups (P<0.05, P<0.01). The numbers of BrdU-positive cells as well as BrdU/NeuN and BrdU/GFAP double-labeled cells were significantly increased in the 3 EA subgroups (P<0.05, P<0.01, P<0.001). CONCLUSION: EA of GV20, GV14 and BL23 can improve the recognition memory ability of mice with radiation-induced brain injury, which may be related to its effect in promoting the proliferation and differentiation of stem cells in the hippocampus.
Assuntos
Lesões Encefálicas , Eletroacupuntura , Células-Tronco Neurais , Animais , Diferenciação Celular , Proliferação de Células , Hipocampo , Camundongos , Camundongos Endogâmicos C57BLRESUMO
The present study was aimed to investigate the effects and mechanisms of electro-acupuncture (EA) on proliferation and differentiation of neural stem cells in the hippocampus of C57 mice exposed to different doses of X-ray radiation. Thirty-day-old C57BL/6J mice were randomly divided into control, irradiation, and EA groups. The control group was not treated with irradiation. The irradiation groups were exposed to different doses of X-ray (4, 8 or 16 Gy) for 10 min. The EA groups were electro-acupunctured at Baihui, Fengfu and bilateral Shenyu for 3 courses of treatment after X-ray radiation. Immunohistochemistry was used to evaluate proliferation and differentiation of the hippocampal neural stem cell. RT-PCR and Western blot were used to detect mRNA and protein expressions of Notch1 and Mash1 in the hippocampus, respectively. The results showed that, compared with the control group, the numbers of BrdU positive cells (4, 8 Gy subgroup) and BrdU/NeuN double-labeling positive cells (3 dose subgroups) were decreased significantly in the irradiation group, but the above changes could be reversed by EA. Compared with the control group, the number of BrdU/GFAP double-labeling positive cells in each dose subgroup of irradiation group was decreased significantly, while EA could reverse the change of 4 and 8 Gy dose subgroups. In addition, compared with the control group, the expression levels of Notch1 mRNA and protein in hippocampus were up-regulated, and the expression levels of Mash1 mRNA and protein were significantly decreased in each dose subgroup of irradiation group. Compared with irradiation group, the expression levels of Notch1 mRNA and protein in hippocampus of EA group were decreased significantly in each dose subgroup, and the expression levels of Mash1 mRNA and protein were increased significantly in 4 and 8 Gy subgroups. These results suggest that irradiation affects the proliferation and differentiation of neural stem cells in hippocampus of mice, whereas EA may significantly increase the proliferation and differentiation of hippocampal neural stem cells via the regulation of Notch signaling pathway.
Assuntos
Diferenciação Celular , Proliferação de Células , Eletroacupuntura , Células-Tronco Neurais/citologia , Raios X/efeitos adversos , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Hipocampo/citologia , Hipocampo/efeitos da radiação , Camundongos Endogâmicos C57BL , Células-Tronco Neurais/efeitos da radiação , Distribuição Aleatória , Receptor Notch1/metabolismoRESUMO
Andrographolide is a medical herbal compound with documented anti-inflammatory activity and therapeutic efficacy in animal models of Alzheimer's disease, traumatic brain injury, and ischemic stroke. The present study examined the potential therapeutic effects of andrographolide on chronic cerebral hypoperfusion (CCH)-induced hippocampal neuronal damage and cognitive dysfunction. A CCH model was established in male Sprague Dawley (SD) rats using 2-vessel occlusion (2VO). After 4 weeks of CCH, spatial learning and memory were assessed in the Morris water maze and structural damage to the hippocampus by hematoxylin and eosin (HE) staining. Astrocyte activation was examined by immunohistochemical staining and Western blotting for glial fibrillary acid protein (GFAP), while expression levels of the pro-inflammatory cytokine-tumor necrosis factor alpha (TNF-α) and interleukin 1 beta (IL-1ß), the apoptosis effector cysteinyl aspartate specific proteinase-3 (caspase-3), and the neuroprotectant brain-derived neurotrophic factor (BDNF) and the TrkB receptor were estimated by Western blotting. After 4 weeks of CCH, the hippocampus of 2VO rats exhibited marked neurodegeneration as well as elevated GFAP, TNF-α, IL-1ß, and caspase-3 compared to Sham controls. In addition, spatial learning was impaired compared to Sham controls. Andrographolide treatment during CCH suppressed astrocyte activation as evidenced by reduced GFAP expression, enhanced expression of BDNF and TrkB, improved impaired spatial learning and memory, and reversed upregulated TNF-α, IL-1ß, and caspase-3 expression. These results reveal a potential neuroprotective effect of andrographolide on hippocampal neuronal damage and cognitive impairment from CCH due to suppression of astrocyte activation and enhancement of BDNF-TrkB signaling.
Assuntos
Isquemia Encefálica/tratamento farmacológico , Diterpenos/farmacologia , Transtornos da Memória/tratamento farmacológico , Fármacos Neuroprotetores/farmacologia , Animais , Apoptose/efeitos dos fármacos , Astrócitos/efeitos dos fármacos , Astrócitos/metabolismo , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Disfunção Cognitiva/tratamento farmacológico , Modelos Animais de Doenças , Hipocampo/efeitos dos fármacos , Hipocampo/patologia , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Memória Espacial/efeitos dos fármacosRESUMO
URB597 (URB) has therapeutic potential for treating chronic cerebral hypoperfusion (CCH)-induced neuronal death. The present study investigated the protective effects of URB on autopahgy and mitophagy in a CCH model as well as the underlying mechanisms. The ultrastructural changes were examined by electron microscopy. The mitochondrial membrane potential was assessed by immunofluorescence. The expressions of autophagy-related proteins (beclin-1, p62, and LC3), lysosome-related proteins (CTSD and LAMP1), and mitophagy-related proteins (BNIP3, cyt C and parkin) were evaluated by western blotting, and the interaction of beclin-1 and Bcl-2 were determined by immunoprecipitation. CCH significantly decreased the protein expression of p62, CTSD, and LAMP1 and increased the protein expression of beclin-1, parkin, and BNIP3, the LC3-II to LC3-I ratio, and the release of cyt C from mitochondria to cytoplasm. Furthermore, CCH induced the accumulation of ubiquitinated proteins in PSDs. However, URB significantly reversed these results. Besides, URB significantly inhibited the beclin-1 from beclin-1/Bcl-2 complex to whole-cell lysates. The above results indicate that URB could inhibit impaired autophagy degradation and the disruption of beclin-1/Bcl-2 complex and subsequently cut off BNIP3-cyt C- and parkin-required mitophagy, finally preventing the abnormal excessive autophagy and mitophagy. These findings provide new insights that URB is a promising agent for therapeutic management of CCH.
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Autofagia/efeitos dos fármacos , Benzamidas/farmacologia , Isquemia Encefálica/patologia , Carbamatos/farmacologia , Mitofagia/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Adenilato Quinase/metabolismo , Animais , Autofagossomos/efeitos dos fármacos , Autofagossomos/metabolismo , Autofagossomos/ultraestrutura , Proteína Beclina-1/metabolismo , Hipocampo/patologia , Lisossomos/efeitos dos fármacos , Lisossomos/metabolismo , Masculino , Fusão de Membrana/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Mitocôndrias/ultraestrutura , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Neurônios/ultraestrutura , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ratos Sprague-Dawley , Sinapses/efeitos dos fármacos , Sinapses/patologia , Sinapses/ultraestrutura , Serina-Treonina Quinases TOR/metabolismoRESUMO
Vascular dementia (VaD) is characteristic of chronic brain ischemia and progressive memory decline, which has a high incidence in the elderly. However, there are no effective treatments for VaD, and the underlying mechanism of its pathogenesis remains unclear. This study investigated the effects of a synthetic cannabinoid receptor agonist WIN55,212-2 (WIN) on VaD, and molecular mechanisms of the effects. VaD model was induced by 2-vessel occlusion (2VO). Spatial reference learning was evaluated by the Morris water maze, and recognition memory was assessed using the novel object recognition test. Autophagy-related proteins [microtubule-associated protein 1 light chain 3 (LC-3) and Beclin-1] were examined by immunohistochemistry and Western blot. Caspase-3 was detected by Western blot. Inflammatory factors, tumor necrosis factor alpha (TNF-α) and interleukin 1 beta (IL-1ß), were estimated by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. VaD increased the levels of LC-3, Beclin-1, and inflammatory factors, which were reversed by chronic treatment with WIN. WIN decreased the expression of Capase-3, and improved the learning and memory impairment of VaD rats. These data indicate that WIN exerts a neuroprotective effect on the cognitive deficits of VaD rats, which may be associated with the suppression of excessive autophagy and inflammation.
Assuntos
Autofagia/efeitos dos fármacos , Benzoxazinas/uso terapêutico , Agonistas de Receptores de Canabinoides/uso terapêutico , Disfunção Cognitiva/tratamento farmacológico , Demência Vascular/tratamento farmacológico , Morfolinas/uso terapêutico , Naftalenos/uso terapêutico , Fármacos Neuroprotetores/uso terapêutico , Animais , Autofagia/fisiologia , Benzoxazinas/farmacologia , Agonistas de Receptores de Canabinoides/farmacologia , Disfunção Cognitiva/patologia , Disfunção Cognitiva/psicologia , Demência Vascular/patologia , Demência Vascular/psicologia , Modelos Animais de Doenças , Inflamação/tratamento farmacológico , Inflamação/patologia , Inflamação/psicologia , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Aprendizagem em Labirinto/fisiologia , Morfolinas/farmacologia , Naftalenos/farmacologia , Fármacos Neuroprotetores/farmacologia , Ratos , Ratos Sprague-DawleyRESUMO
The purpose of this study was to explore the effects of different concentrations of calcitonin gene-related peptide (CGRP) on long-term potentiation (LTP) in the hippocampus of mice. C57BL/6J mice (30 days old) were randomly divided into control group, three CGRP groups, and CGRP + CGRP8-37 group (10 mice for each group). Different concentrations of CGRP (50, 100 and 200 nmol/L) were given to the hippocampal slices of mice. The presynaptic release of neurotransmitters and the induction of LTP were measured by extracellular field recording techniques. The result showed that different concentrations of CGRP did not affect the presynaptic release of neurotransmitters, but 100 and 200 nmol/L CGRP increased the amplitude of LTP induced in the hippocampus of mice. This facilitation effect of CGRP was blocked by its specific antagonist CGRP8-37. These results suggest that CGRP dose-dependently facilitates the induction of LTP in the hippocampus of mice through its specific receptor.
Assuntos
Peptídeo Relacionado com Gene de Calcitonina/farmacologia , Hipocampo/efeitos dos fármacos , Potenciação de Longa Duração , Fragmentos de Peptídeos/farmacologia , Animais , Antagonistas do Receptor do Peptídeo Relacionado ao Gene de Calcitonina , Camundongos , Camundongos Endogâmicos C57BL , Neurotransmissores/análiseRESUMO
The present study explored the protective effects of cannabinoid receptor agonist WIN55,212-2 (WIN) and fatty acid amide hydrolase inhibitor URB597 (URB) against neuroinflammation in rats with chronic cerebral hypoperfusion (CCH). Activated microglia, astrocytes, and nuclear factor kappa B (NF-κB) p65-positive cells were measured by immunofluorescence. Reactive oxygen species (ROS) was assessed by dihydroethidium staining. The protein levels of cluster of differentiation molecule 11b (OX-42), glial fibrillary acidic protein (GFAP), NF-κB p65, inhibitor of kappa B alpha (IκB-a), IκB kinase a/ß (IKK a/ß), phosphorylated IKK a/ß (p-IKK a/ß), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-α, and interleukin-1ß (IL-1ß) were examined by western blotting or enzyme-linked immunosorbent assay. All the protein levels of OX-42, GFAP, TNF-a, IL-1ß, COX-2, and iNOS are increased in CCH rats. WIN and URB downregulated the levels of OX-42, GFAP, TNF-α, IL-1ß, COX-2 and iNOS and inhibited CCH-induced ROS accumulation in CCH rats, indicating that WIN and URB might exert their neuroprotective effects by inhibiting the neuroinflammatory response. In addition, the NF-κB signaling pathway was activated by CCH in frontal cortex and hippocampus, while the aforementioned changes were reversed by WIN and URB treatment. These findings suggest that WIN and URB treatment ameliorated CCH-induced neuroinflammation through inhibition of the classical pathway of NF-κB activation, resulting in mitigation of chronic ischemic injury.
Assuntos
Amidoidrolases/antagonistas & inibidores , Anti-Inflamatórios/uso terapêutico , Benzamidas/uso terapêutico , Benzoxazinas/uso terapêutico , Agonistas de Receptores de Canabinoides/uso terapêutico , Carbamatos/uso terapêutico , Transtornos Cerebrovasculares/tratamento farmacológico , Morfolinas/uso terapêutico , NF-kappa B/antagonistas & inibidores , Naftalenos/uso terapêutico , Neurite (Inflamação)/tratamento farmacológico , Transdução de Sinais/efeitos dos fármacos , Animais , Anti-Inflamatórios/farmacologia , Benzamidas/farmacologia , Benzoxazinas/farmacologia , Agonistas de Receptores de Canabinoides/farmacologia , Carbamatos/farmacologia , Transtornos Cerebrovasculares/fisiopatologia , Endocanabinoides/fisiologia , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Hipocampo/patologia , Ativação de Macrófagos/efeitos dos fármacos , Masculino , Morfolinas/farmacologia , Naftalenos/farmacologia , Neurite (Inflamação)/fisiopatologia , Córtex Pré-Frontal/efeitos dos fármacos , Córtex Pré-Frontal/metabolismo , Córtex Pré-Frontal/patologia , Ratos , Ratos Sprague-Dawley , Fator de Transcrição RelA/efeitos dos fármacosRESUMO
The present study further investigated the protective effects of cannabinoid receptor agonist WIN55,212-2 (WIN) and fatty acid amide hydrolase (FAAH) inhibitor URB597 (URB) on chronic cerebral hypoperfusion (CCH)-induced cognitive impairment in rats. Spatial learning and memory were assessed with the Morris water maze and by measuring Long-term potentiation. The expression of microtubule-associated protein-2 (MAP)-2, growth-associated protein-43 (GAP)-43, synaptophysin, cannabinoid receptor 1 (CB1), brain-derived neurotrophic factor (BDNF), FAAH, N-acylphosphatidylethanolamine phospholipase D(NAPE-PLD) and monoacyl glycerol lipase (MGL) as well as phosphoinositide 3-kinase (PI3K)/AKT signaling pathway molecules and downstream targets including AKT, phosphorylated (p-)AKT, cyclic AMP response element- binding protein (CREB), p-CREB, Bcl-2-associated death protein (BAD), p-BAD, glycogen synthase kinase (GSK)-3ß, p-GSK-3ß, forkhead box protein (FOXO) 3A and p-FOXO3A was determined by western blotting. WIN and URB treatment improved learning and memory performance, effects that were abolished by co-administration of the PI3K/AKT inhibitor LY294002. Moreover, WIN and URB reversed the decreases in MAP-2 and synaptophysin expression resulting from CCH, and stimulated BDNF and CB1 expression as well as CREB, FOXO3A, GSK-3ß, and BAD phosphorylation, confirming that WIN and URB mediate neuroprotection by preventing neuronal apoptosis and improving cognition via PI3K/AKT signaling. These findings suggest that WIN and URB are promising agents for therapeutic management of CCH.
Assuntos
Benzamidas/farmacologia , Benzoxazinas/farmacologia , Agonistas de Receptores de Canabinoides/farmacologia , Carbamatos/farmacologia , Potenciação de Longa Duração/efeitos dos fármacos , Oxigenases de Função Mista/antagonistas & inibidores , Morfolinas/farmacologia , Naftalenos/farmacologia , Receptores de Canabinoides/efeitos dos fármacos , Animais , Disfunção Cognitiva/tratamento farmacológico , Ácidos Graxos/metabolismo , Quinase 3 da Glicogênio Sintase/metabolismo , Masculino , Aprendizagem em Labirinto/fisiologia , Fármacos Neuroprotetores/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacosRESUMO
Detected unruptured intracranial aneurysms (UIA) are becoming more common with the increased utilization of CT angiography, MR angiography and digital subtraction angiography. A proportion of patients with UIA remain untreated. We investigated to assess cognitive function, depression, anxiety and quality of life (QoL) in Chinese patients with untreated UIA. Thirty one Chinese patients with untreated UIA and 25 healthy controls were identified and matched for variables including age, sex, and living area. Cognitive function was evaluated with the Montreal Cognitive Assessment (MoCA). Depression, anxiety and QoL were screened with the Self-Rating Depression Scale, Self-Rating Anxiety Scale, and Short Form-36, respectively. Non-parametric tests were used for comparisons between groups. No patient had cognitive dysfunction at 1 month or 1 year after detection of UIA. However, a significant decrease of overall MoCA subscores was found in 30 (97%) of 31 patients 5 years after UIA discovery, suggestive of mild cognitive impairment. A significant decrease in depression and anxiety was found in patients over time. QoL in patients was reduced most prominently in psychosocial function and social activities 1 year after detection of UIA, but these improved to within normal limits at the end of the follow-up period. For Chinese patients with untreated UIA, depression, anxiety and reduced QoL may be short-term complications. Mild cognitive impairment may be a long-term complication.
Assuntos
Ansiedade , Cognição , Depressão , Aneurisma Intracraniano/psicologia , Qualidade de Vida , Adulto , Ansiedade/epidemiologia , China/epidemiologia , Estudos de Coortes , Depressão/epidemiologia , Feminino , Seguimentos , Humanos , Entrevistas como Assunto , Aneurisma Intracraniano/epidemiologia , Masculino , Pessoa de Meia-Idade , Testes Neuropsicológicos , Escalas de Graduação Psiquiátrica , Inquéritos e QuestionáriosRESUMO
Aneurysmal subarachnoid hemorrhage (aSAH)-induced cerebral vasospasm and delayed ischemic neurological deficit (DIND) are the major causes of morbidity and mortality in patients with aSAH. The effects of statins-use for patients with aSAH remain controversial. Here,a total of 249 patients from six randomized controlled trials(RCTs) were subjected to meta-analysis. No significant decrease was found in the incidence of vasospasm(RR, 0.80; 95% CI, 0.54-1.17), with substantial heterogeneity (I(2) = 49%, P = 0.08), which was verified by the further sensitivity analysis and subgroup meta-analysis. Furthermore, no significant difference was presented in the incidence of poor neurological outcome(RR, 0.94; 95% CI, 0.77-1.16), and potential side effects(RR, 2.49; 95% CI, 0.75-8.33). Nevertheless, significant difference was reported in the occurrence of DIND(RR, 0.58; 95% CI, 0.37-0.92) and mortality(RR, 0.30; 95% CI, 0.14-0.64). At present, although statins-use in the patients with aSAH should not be considered standard care at present, statins-use may have the potential effects in the prevention of mortality in patients with aSAH.
Assuntos
Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Hemorragia Subaracnóidea/epidemiologia , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/efeitos adversos , Incidência , Razão de Chances , Prognóstico , Viés de Publicação , Ensaios Clínicos Controlados Aleatórios como Assunto , Hemorragia Subaracnóidea/complicações , Resultado do Tratamento , Vasoespasmo Intracraniano/epidemiologia , Vasoespasmo Intracraniano/etiologia , Vasoespasmo Intracraniano/prevenção & controleRESUMO
BACKGROUND: Although international guideline recommended routine intracranial pressure (ICP) monitoring for patients with severe traumatic brain injury(TBI), there were conflicting outcomes attributable to ICP monitoring according to the published studies. Hence, we conducted a meta-analysis to evaluate the efficacy and safety of ICP monitoring in patients with TBI. METHODS: Based on previous reviews, PubMed and two Chinese databases (Wangfang and VIP) were further searched to identify eligible studies. The primary outcome was mortality. Secondary outcomes included unfavourable outcome, adverse events, length of ICU stay and length of hospital stay. Weighted mean difference (WMD), odds ratio (OR) and 95% confidence intervals (CIs) were calculated and pooled using fixed-effects or random-effects model. RESULTS: two randomized controlled trials (RCTs) and seven cohort studies involving 11,038 patients met the inclusion criteria. ICP monitoring was not associated with a significant reduction in mortality (OR, 1.16; 95% CI, 0.87-1.54), with substantial heterogeneity (I(2)â= 80%, P<0.00001), which was verified by the sensitivity analyses. No significant difference was found in the occurrence of unfavourable outcome (OR, 1.40; 95% CI, 0.99-1.98; I(2)â= 4%, P = 0.35) and adverse events (OR, 1.04; 95% CI, 0.64-1.70; I(2)â= 78%, P = 0.03). However, we should be cautious to the result of adverse events because of the substantial heterogeneity in the comparison. Furthermore, longer ICU and hospital stay were the consistent tendency according to the pooled studies. CONCLUSIONS: No benefit was found in patients with TBI who underwent ICP monitoring. Considering substantial clinical heterogeneity, further large sample size RCTs are needed to confirm the current findings.