scholarly journals Clarified Açaí (Euterpe oleracea) Juice as an Anticonvulsant Agent: In Vitro Mechanistic Study of GABAergic Targets

2018 ◽  
Vol 2018 ◽  
pp. 1-6 ◽  
Author(s):  
Gabriela P. F. Arrifano ◽  
Mathieu P. Lichtenstein ◽  
José Rogério Souza-Monteiro ◽  
Marcelo Farina ◽  
Hervé Rogez ◽  
...  
Keyword(s):  
Cortical Neurons ◽  
Molecular Mechanisms ◽  
Primary Cultures ◽  
Mechanistic Study ◽  
Gaba Uptake ◽  
Euterpe Oleracea ◽  
Adequate Treatment ◽  
Gabaergic Neurotransmission

Seizures affect about 50 million people around the world. Approximately 30% of seizures are refractory to the current pharmacological arsenal, so, the pursuit of new therapeutic alternatives is essential. Clarified Euterpe oleracea (EO) juice showed anticonvulsant properties similar to diazepam in an in vivo model with pentylenetetrazol, a GABAA receptor blocker. This study investigated the effects of EO on the main GABAergic targets for anticonvulsant drugs, analyzing the effect on the GABA receptor’s benzodiazepine and picrotoxinin binding sites and the GABA uptake. Primary cultures of cortical neurons and astrocytes were treated with EO (0–25%) for up to 90 min. [3H]Flunitrazepam and [3H]TBOB binding, [3H]GABA uptake, cell viability, and morphology were assayed. Nonlethal concentrations of EO increased agonist binding and decreased antagonist binding in cortical neurons. Low concentrations significantly inhibited GABA uptake, especially in astrocytes, suggesting an accumulation of endogenous GABA in the synaptic cleft. The results demonstrate, for the first time, that EO can improve GABAergic neurotransmission via interactions with GABAA receptor and modulation of GABA uptake. Understanding these molecular mechanisms will help in the treatment of seizures and epilepsy, especially in developing countries where geographic isolation and low purchasing power are the main barriers to access to adequate treatment.

scholarly journals Synthesis of New Water Soluble β-Cyclodextrin@Curcumin Conjugates and In Vitro Safety Evaluation in Primary Cultures of Rat Cortical Neurons

2021 ◽  
Vol 22 (6) ◽  
pp. 3255
Author(s):  
Amina Ben Mihoub ◽  
Samir Acherar ◽  
Céline Frochot ◽  
Catherine Malaplate ◽  
Frances T. Yen ◽  
...  
Keyword(s):  
Neurodegenerative Diseases ◽  
Cortical Neurons ◽  
Water Solubility ◽  
Photophysical Properties ◽  
Primary Cultures ◽  
Average Diameter ◽  
Water Soluble ◽  
Cell Toxicity

Self-aggregation of Curcumin (Cur) in aqueous biological environment decreases its bioavailability and in vivo therapeutic efficacy, which hampers its clinical use as candidate for reducing risk of neurodegenerative diseases. Here, we focused on the design of new Cur- β-Cyclodextrin nanoconjugates to improve the solubility and reduce cell toxicity of Cur. In this study, we described the synthesis, structural characterization, photophysical properties and neuron cell toxicity of two new water soluble β-CD/Cur nanoconjugates as new strategy for reducing risks of neurodegenerative diseases. Cur was coupled to one or two β-CD molecules via triazole rings using CuAAC click chemistry strategy to yield β-CD@Cur and (β-CD)2@Cur nanoconjugates, respectively. The synthesized nanoconjugates were found to be able to self-assemble in aqueous condition and form nano-aggregates of an average diameter size of around 35 and 120 nm for β-CD@Cur and (β-CD)2@Cur, respectively. The photophysical properties, water solubility and cell toxicity on rat embryonic cortical neurons of the designed nanoconjugates were investigated and compared to that of Cur alone. The findings revealed that both new nanoconjugates displayed better water solubility and in vitro biocompatibility than Cur alone, thus making it possible to envisage their use as future nano-systems for the prevention or risk reduction of neurodegenerative diseases.

scholarly journals In Vitro and In Vivo Models to Study Nephropathic Cystinosis

Cells ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 6
Author(s):  
Pang Yuk Cheung ◽  
Patrick T. Harrison ◽  
Alan J. Davidson ◽  
Jennifer A. Hollywood
Keyword(s):  
Cell Lines ◽  
Molecular Mechanisms ◽  
Primary Cultures ◽  
Model Systems ◽  
In Vivo Model ◽  
Nephropathic Cystinosis ◽  
In Vivo Models ◽  
Induced Pluripotent

The development over the past 50 years of a variety of cell lines and animal models has provided valuable tools to understand the pathophysiology of nephropathic cystinosis. Primary cultures from patient biopsies have been instrumental in determining the primary cause of cystine accumulation in the lysosomes. Immortalised cell lines have been established using different gene constructs and have revealed a wealth of knowledge concerning the molecular mechanisms that underlie cystinosis. More recently, the generation of induced pluripotent stem cells, kidney organoids and tubuloids have helped bridge the gap between in vitro and in vivo model systems. The development of genetically modified mice and rats have made it possible to explore the cystinotic phenotype in an in vivo setting. All of these models have helped shape our understanding of cystinosis and have led to the conclusion that cystine accumulation is not the only pathology that needs targeting in this multisystemic disease. This review provides an overview of the in vitro and in vivo models available to study cystinosis, how well they recapitulate the disease phenotype, and their limitations.

scholarly journals MKK3 sustains cell proliferation and survival through p38DELTA MAPK activation in colorectal cancer

2019 ◽  
Vol 10 (11) ◽  
Author(s):  
Lorenzo Stramucci ◽  
Angelina Pranteda ◽  
Arianna Stravato ◽  
Carla Azzurra Amoreo ◽  
Annarita Pennetti ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Map Kinases ◽  
Molecular Mechanisms ◽  
Malignant Tumors ◽  
Mapk Pathway ◽  
Primary Cultures ◽  
Mitogen Activated Protein Kinase ◽  
Antitumor Effects

Abstract Colorectal cancer (CRC) is one of the most common malignant tumors worldwide and understanding its underlying molecular mechanisms is crucial for the development of therapeutic strategies. The mitogen-activated protein kinase-kinase 3 (MKK3) is a specific activator of p38 MAP kinases (p38 MAPKs), which contributes to the regulation of several cellular functions, such as proliferation, differentiation, apoptosis as well as response to drugs. At present, the exact MKK3/p38 MAPK pathway contribution in cancer is heavily debated because of its pleiotropic function. In this work, we retrospectively explored the prognostic and pathobiologic relevance of MKK3 in a cohort of CRC patients and assessed MKK3 molecular functions in a panel of CRC lines and colonocytes primary cultures. We found increased MKK3 levels in late-stage CRC patients which correlated with shorter overall survival. Herein, we report that the MKK3 targeting by inducible RNA interference univocally exerts antitumor effects in CRC lines but not in primary colonocytes. While MKK3 depletion per se affects growth and survival by induction of sustained autophagy and death in some CRC lines, it potentiates response to chemotherapeutic drug 5-fluorouracil (5-FU) in all of the tested CRC lines in vitro. Here, we demonstrate for the first time that in CRC the MKK3 specifically activates p38delta MAPK isoform to sustain prosurvival signaling and that such effect is exacerbated upon 5-FU challenge. Indeed, p38delta MAPK silencing recapitulates MKK3 depletion effects in CRC cells in vitro and in vivo. Overall, our data identified a molecular mechanism through which MKK3 supports proliferation and survival signaling in CRC, further supporting MKK3 as a novel and extremely attractive therapeutic target for the development of promising strategies for the management of CRC patients.

scholarly journals Long-Lasting Changes in Glial Cells Isolated From Rats Subjected to the Valproic Acid Model of Autism Spectrum Disorder

2021 ◽  
Vol 12 ◽  
Author(s):  
Marianela Evelyn Traetta ◽  
Nonthué Alejandra Uccelli ◽  
Sandra Cristina Zárate ◽  
Dante Gómez Cuautle ◽  
Alberto Javier Ramos ◽  
...  
Keyword(s):  
Autism Spectrum Disorder ◽  
Valproic Acid ◽  
Cortical Neurons ◽  
Primary Cultures ◽  
Autism Spectrum ◽  
Spectrum Disorder ◽  
Saline Control ◽  
Synaptic Changes

Synaptic alterations concomitant with neuroinflammation have been described in patients and experimental models of autism spectrum disorder (ASD). However, the role of microglia and astroglia in relation to synaptic changes is poorly understood. Male Wistar rats prenatally exposed to valproic acid (VPA, 450 mg/kg, i.p.) or saline (control) at embryonic day 10.5 were used to study synapses, microglia, and astroglia in the prefrontal cortex (PFC) at postnatal days 3 and 35 (PND3 and PND35). Primary cultures of cortical neurons, microglia, and astroglia isolated from control and VPA animals were used to study each cell type individually, neuron-microglia and microglia-astroglia crosstalk. In the PFC of VPA rats, synaptic changes characterized by an increase in the number of excitatory synapses were evidenced at PND3 and persisted until PND35. At PND3, microglia and astroglia from VPA animals were morphologically similar to those of age-matched controls, whereas at PND35, reactive microgliosis and astrogliosis were observed in the PFC of VPA animals. Cortical neurons isolated from VPA rats mimicked in vitro the synaptic pattern seen in vivo. Cortical microglia and astroglia isolated from VPA animals exhibited reactive morphology, increased pro-inflammatory cytokines, and a compromised miRNA processing machinery. Microglia from VPA animals also showed resistance to a phagocytic challenge. In the presence of neurons from VPA animals, microglia isolated from VPA rats revealed a non-reactive morphology and promoted neurite outgrowth, while microglia from control animals displayed a reactive profile and promoted dendritic retraction. In microglia-astroglia co-cultures, microglia from VPA animals displayed a reactive profile and exacerbated astrocyte reactivity. Our study indicates that cortical microglia from VPA animals are insensitive or adapted to neuronal cues expressed by neurons from VPA animals. Further, long-term in vivo microgliosis could be the result of altered microglia-astroglia crosstalk in VPA animals. Thus, our study highlights cortical microglia-astroglia communication as a new mechanism implicated in neuroinflammation in ASD; consequently, we propose that this crosstalk is a potential target for interventions in this disorder.

scholarly journals COX5A over-expression protects cortical neurons from hypoxic ischemic injury in neonatal rats associated with TPI up-regulation

2020 ◽  
Author(s):  
Ya Jiang ◽  
Xue Bai ◽  
Qiong Zhao ◽  
Mohammed AL Hawwas ◽  
Yuan Jin ◽  
...  
Keyword(s):  
Cortical Neurons ◽  
Molecular Mechanisms ◽  
Brain Infarction ◽  
Glucose Deprivation ◽  
Immunofluorescent Staining ◽  
Neonatal Rats ◽  
Over Expression ◽  
Qrt Pcr

Abstract Background: Neonatal hypoxic-ischemic encephalopathy (HIE) is a destructive condition that constitutes a main cause of death in newborns. However, the underlying molecular mechanisms in brain damage are still not fully elucidated. Results: Here, we established hypoxic-ischemic (HI) injury and primary cortical neurons subjected to oxygen-glucose deprivation (OGD) to mimic HIE model in-vivo and in - vitro . Zea-longa scores, Triphenyte-trazoliumchloride (TTC) staining the Terminal Deoxynucleotidyl Transferased Utp Nick End Labeling (tunel) and immunofluorescent staining were used to detect the neurological injuries after HI. Then the expression of Cytochrome c oxidase subunit 5a (COX5A) was determined by immunohistochemistry, western blotting (WB) and quantitative real time Polymerase Chain Reaction (qRT-PCR) techniques. Moreover, HSV-mediated COX5A over-expression virus was administrated in - vitro to explore the role of COX5A in OGD neurons. Subsequently, the underlying mechanism was predicted by GeneMANIA and confirmed by WB and qRT-PCR. The results showed that HI induced a severe behavioral dysfunction, brain infarction, and cell apoptosis as well as obvious neuron loss in neonatal rats, in corresponding to the decrease on the expression of COX5A in both sides of the brain . What’s more, COX5A over-expression significantly promoted the neuron survival, reduced the apoptosis rate, and markedly increased the neurites length after OGD. Moreover, Triosephosephate isomerase (TPI) was predicted as physical interactions with COX5A, and COX5A over-expression largely increased the expressional level of TPI. Conclusions: Together, these data suggest that COX5A plays an important role in promoting neurological recovery after HI, and this process is related to TPI up-regulation.

scholarly journals LncRNA NKX2-1-AS1 As ceRNA Promotes Tumor Progression and Angiogenesis by Upregulating SERPINE1 Expression and Activating VEGFR-2 Signaling Pathway in Gastric Cancer

2020 ◽  
Author(s):  
Fei Teng ◽  
Juxiang Zhang ◽  
Yi Chen ◽  
Xiaodong Shen ◽  
Yanjiao Guo ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Tumor Progression ◽  
Molecular Mechanisms ◽  
Luciferase Assay ◽  
Luciferase Reporter ◽  
Mechanistic Study ◽  
Tumor Tissues ◽  
Vegfr2 Signaling

Abstract Background: Recent evidence indicated that the lncRNA NKX2-1-AS1 (NKX2-1 antisense RNA 1) plays an important role in cancer progression and metastasis. However, the associated molecular mechanisms of NKX2-1-AS1 in GC are still unclear. Methods: To determine the target of the study by bioinformatic analysis. NKX2-1-AS1 expression was measured in paired tumor and non-tumor tissues of 178 GC patients, by quantitative reverse transcription PCR. The in vitro and in vivo biological functions of NKX2-1-AS1 were examined by loss-of-function and gain-of-function experiments. The potential mechanisms of this competing endogenous RNA (ceRNA) were elucidated using dual-luciferase reporter assay, quantitative PCR, Western blot, and fluorescence in situ hybridization (FISH). Results: NKX2-1-AS1 expression was upregulated in GC cell lines and tumor tissues, which was correlated with tumor progression and enhanced angiogenesis. Functionally, NKX2-1-AS1 overexpression promoted GC cell proliferation, metastasis, invasion, and angiogenesis, while NKX2-1-AS1 downregulation reversed these effects, both in vitro and in vivo. Bioinformatics analysis and dual-luciferase assay showed that the microRNA miR-145-5p is a direct target of NKX2-1-AS1, and that NKX2-1-AS1 acts as a ceRNA that regulates angiogenesis in the context of GC. The mechanistic study revealed that miR-145-5p specifically targets serpin family E member 1 (SERPINE1), and that the complex NKX2-1-AS1/miR-145-5p activates VEGFR2 signaling, via SERPINE1, to promote tumor proliferation and angiogenesis. Conclusion: NKX2-1-AS1 upregulation is associated with tumor cell proliferation, increased angiogenesis, and poor prognosis in GC patients. NKX2-1-AS1 regulates SERPINE1 expression and VEGFR2 signaling by acting as a ceRNA for miR-145-5p.

scholarly journals SETD5 Binding to EP300/HIF1α is Required for Glycolysis in Breast Cancer Stem-Like Cells

2020 ◽  
Author(s):  
Zhaoting Yang ◽  
Huazi Li ◽  
Chengye Zhang ◽  
Nan Che ◽  
Ying Feng ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Molecular Mechanisms ◽  
Hypoxia Inducible Factor ◽  
Lactate Production ◽  
Xenograft Model ◽  
Regulatory Function ◽  
Mechanistic Study ◽  
The Impact

Abstract BackgroundGlycolysis plays a pivotal role in breast cancer stem-like cell reprogramming. The SET-domain containing 5 (SETD5) is a previously uncharacterized member of the histone lysine methyltransferase family. Yet, the molecular mechanisms underlying the promotion of stem-like and glycolysis activation traits of SETD5 have not been elucidated.MethodsBasing on public datasets, we explored clinicopathological and survival analysis of SETD5 on breast cancer (BC) patients. Spheroid formation, transfection experiments and measurement of glucose uptake and lactate production analyzed the regulatory function of SETD5 on glycolysis in breast cancer stem-like cells (BCSC). The impact of SETD5 on tumor growth was studied in a murine xenograft model. Immunohistochemistry, immunofluorescence, western blot, preparation of cytoplasmic and nuclear extracts and co-immunoprecipitation were used to determine the molecular mechanisms of SETD5 in cancer cell glycolysis.ResultsOur data displayed that overexpression of SETD5 in BC tissues is positively associated with progression. SETD5 overexpression is associated with poor post-progression survival in BC patients. SETD5 expression was enriched in spheroid cells. Downregulation of SETD5 significantly decreased BCSC properties and glycolysis in vitro and in vivo. Interestingly, SETD5 and glycolytic enzymes were accumulated in the central hypoxic regions of subcutaneous tumor tissues. Our mechanistic study found that SETD5 binding to EP300/hypoxia-inducible factor 1α (HIF1α) and work as an upstream effector. SETD5 knockdown reduced the expression of HIF1α, hexokinase-2, and 6-phosphofructo-2-kinase in the nucleus after treatment with cobalt chloride (CoCl2), a chemical hypoxia mimetic agent, which activates HIF1α to accumulate in the nucleus. ConclusionSETD5 is required for glycolysis in BCSCs through binding to EP300/HIF1α and could be a potential therapeutic target for BC patients.

Neuroprotective effect of dauricine in cortical neuron culture exposed to hypoxia and hypoglycemia: involvement of correcting perturbed calcium homeostasis

2007 ◽  
Vol 85 (6) ◽  
pp. 621-627 ◽  
Author(s):  
Yan-Hong Li ◽  
Pei-Li Gong
Keyword(s):  
Cerebral Ischemia ◽  
Cortical Neurons ◽  
Focal Cerebral Ischemia ◽  
Neuroprotective Effect ◽  
Primary Cultures ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Method ◽  
Fluorescence Measurements

We have previously reported that dauricine protects brain tissues from focal cerebral ischemia. To corroborate this effect, neurotoxicity due to hypoxia and hypoglycemia was assessed in primary cultures of rat cortical neurons by using a trypan blue exclusion method. To further clarify the mechanism, the intracellular Ca2+ concentration ([Ca2+]i) and mitochondrial membrane potential (ΔΨm) of dissociated rat cortical cells were monitored by fura-2 fluorescence measurements and flow cytometry, respectively. The results showed that 1 and 10 μmol/L dauricine significantly enhanced neuronal survival during 4 h of hypoxia and hypoglycemia. Dauricine inhibited the increase in [Ca2+]i and decrease in ΔΨm induced by 30 min of hypoxia and hypoglycemia. When exploring the pathway, we found that 1 μmol/L dauricine inhibited the [Ca2+]i increase induced by 7.5 nmol/L thapsigargin in either the presence or absence of extracellular Ca2+ and by 1 mmol/L l-glutamate in the presence of extracellular Ca2+. These results suggest that dauricine prevents neuronal loss from ischemia in vitro, which is in accordance with our previous research in vivo. In addition, by inhibiting Ca2+ release from the endoplasmic reticulum and Ca2+ influx from the extracellular space, dauricine suppressed the increase in [Ca2+]i and, subsequently, the decrease in ΔΨm induced by hypoxia and hypoglycemia. This effect may underlie the mechanism of action of dauricine on cerebral ischemia.

scholarly journals Controlled Decompression Attenuates Compressive Injury following Traumatic Brain Injury via TREK-1-Mediated Inhibition of Necroptosis and Neuroinflammation

2021 ◽  
Vol 2021 ◽  
pp. 1-17
Author(s):  
Tao Chen ◽  
Xiao Qian ◽  
Jie Zhu ◽  
Li-Kun Yang ◽  
Yu-Hai Wang
Keyword(s):  
Traumatic Brain Injury ◽  
Brain Injury ◽  
Intracranial Hypertension ◽  
Cortical Neurons ◽  
Molecular Mechanisms ◽  
Neuronal Injury ◽  
K Channel ◽  
Protective Effects

Decompressive craniectomy is an effective strategy to reduce intracranial hypertension after traumatic brain injury (TBI), but it is related to many postoperative complications, such as delayed intracranial hematoma and diffuse brain swelling. Our previous studies have demonstrated that controlled decompression (CDC) surgery attenuates brain injury and reduces the rate of complications after TBI. Here, we investigated the potential molecular mechanisms of CDC in experimental models. The in vitro experiments were performed in a traumatic neuronal injury (TNI) model following compression treatment in primary cultured cortical neurons. We found that compression aggravates TNI-induced neuronal injury, which was significantly attenuated by CDC for 2 h or 3 h. The results of immunocytochemistry showed that CDC reduced neuronal necroptosis and activation of RIP3 induced by TNI and compression, with no effect on RIP1 activity. These protective effects were associated with decreased levels of inflammatory cytokines and preserved intracellular Ca2+ homeostasis. In addition, the expression of the two-pore domain K+ channel TREK-1 and its activity was increased by compression and prolonged by CDC. Treatment with the TREK-1 blockers, spadin or SID1900, could partially prevent the effects of CDC on intracellular Ca2+ metabolism, necroptosis, and neuronal injury following TNI and compression. Using a traumatic intracranial hypertension model in rats, we found that CDC for 20 min or 30 min was effective in alleviating brain edema and locomotor impairment in vivo. CDC significantly inhibited neuronal necroptosis and neuroinflammation and increased TREK-1 activation, and the CDC-induced protection in vivo was attenuated by spadin and SID1900. In summary, CDC is effective in alleviating compressive neuronal injury both in vitro and in vivo, which is associated with the TREK-1-mediated attenuation of intracellular Ca2+ overload, neuronal necroptosis, and neuroinflammation.

Flavonoids as P-gp Inhibitors: A Systematic Review of SARs

2019 ◽  
Vol 26 (25) ◽  
pp. 4799-4831 ◽  
Author(s):  
Jiahua Cui ◽  
Xiaoyang Liu ◽  
Larry M.C. Chow
Keyword(s):  
Molecular Mechanisms ◽  
Metastatic Cancer ◽  
Drug Candidates ◽  
Chemical Structures ◽  
Transporter Family ◽  
Promising Area ◽  
New Generation ◽  
Nontoxic Inhibitors

P-glycoprotein, also known as ABCB1 in the ABC transporter family, confers the simultaneous resistance of metastatic cancer cells towards various anticancer drugs with different targets and diverse chemical structures. The exploration of safe and specific inhibitors of this pump has always been the pursuit of scientists for the past four decades. Naturally occurring flavonoids as benzopyrone derivatives were recognized as a class of nontoxic inhibitors of P-gp. The recent advent of synthetic flavonoid dimer FD18, as a potent P-gp modulator in reversing multidrug resistance both in vitro and in vivo, specifically targeted the pseudodimeric structure of the drug transporter and represented a new generation of inhibitors with high transporter binding affinity and low toxicity. This review concerned the recent updates on the structure-activity relationships of flavonoids as P-gp inhibitors, the molecular mechanisms of their action and their ability to overcome P-gp-mediated MDR in preclinical studies. It had crucial implications on the discovery of new drug candidates that modulated the efflux of ABC transporters and also provided some clues for the future development in this promising area.

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