Pharmacological Investigations in Glia Culture Model of Inflammation

Astrocytes and microglia are the main cell population besides neurons in the central nervous system (CNS). Astrocytes support the neuronal network via maintenance of transmitter and ion homeostasis. They are part of the tripartite synapse, composed of pre- and postsynaptic neurons and perisynaptic astrocytic processes as a functional unit. There is an increasing evidence that astroglia are involved in the pathophysiology of CNS disorders such as epilepsy, autoimmune CNS diseases or neuropsychiatric disorders, especially with regard to glia-mediated inflammation. In addition to astrocytes, investigations on microglial cells, the main immune cells of the CNS, offer a whole network approach leading to better understanding of non-neuronal cells and their pathological role in CNS diseases and treatment. An in vitro astrocyte-microglia co-culture model of inflammation was developed by Faustmann et al. (2003), which allows to study the endogenous inflammatory reaction and the cytokine expression under drugs in a differentiated manner. Commonly used antiepileptic drugs (e.g., levetiracetam, valproic acid, carbamazepine, phenytoin, and gabapentin), immunomodulatory drugs (e.g., dexamethasone and interferon-beta), hormones and psychotropic drugs (e.g., venlafaxine) were already investigated, contributing to better understanding mechanisms of actions of CNS drugs and their pro- or anti-inflammatory properties concerning glial cells. Furthermore, the effects of drugs on glial cell viability, proliferation and astrocytic network were demonstrated. The in vitro astrocyte-microglia co-culture model of inflammation proved to be suitable as unique in vitro model for pharmacological investigations on astrocytes and microglia with future potential (e.g., cancer drugs, antidementia drugs, and toxicologic studies).

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Progress and Promise of Nur77-based Therapeutics for Central Nervous System Disorders

Current Neuropharmacology ◽

10.2174/1570159x18666200606231723 ◽

2020 ◽

Vol 18 ◽

Author(s):

Lu Liu ◽

Di Ma ◽

La Zhuo ◽

Xinyuan Pang ◽

Jiulin You ◽

...

Keyword(s):

Central Nervous System ◽

Nervous System ◽

Neurological Disorders ◽

Cns Diseases ◽

Multiple Systems ◽

Mitochondrial Homeostasis ◽

Natural Ligand ◽

The Central Nervous System

: Nur77 belongs to the NR4A subgroup of the nuclear receptor superfamily. Unlike other nuclear receptors, a natural ligand for Nur77 has not been identified yet. However, a few small molecules can interact with this receptor and induce a conformational change to mediate its activity. The expression and activation of Nur77 can be rapidly increased using various physiological and pathological stimuli. In vivo and in vitro studies have demonstrated its regulatory role in tissues and cells of multiple systems by means of participation in cell differentiation, apoptosis, metabolism, mitochondrial homeostasis, and other processes. Although research on Nur77 in the pathophysiology of the central nervous system (CNS) is currently limited, the present data support the fact that Nur77 is involved in many neurological disorders such as stroke, multiple sclerosis, Parkinson’s disease. This indicates that activation of Nur77 has considerable potential in treating these diseases. This review summarizes the regulatory mechanisms of Nur77 in CNS diseases and presents available evidence for its potential as a targeted therapy, especially for cerebrovascular and inflammation-related CNS diseases.

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A Primary Neural Cell Culture Model for Neuroinflammation

10.1101/2020.01.23.914226 ◽

2020 ◽

Author(s):

Noah Goshi ◽

Rhianna K. Morgan ◽

Pamela J. Lein ◽

Erkin Seker

Keyword(s):

Cell Types ◽

Mechanical Trauma ◽

Cortical Cells ◽

Neuroinflammatory Response ◽

Physiological Relevance ◽

Culture Model ◽

Culture Models ◽

The Central Nervous System

AbstractInteractions between neurons, astrocytes and microglia critically influence neuroinflammatory responses to insult in the central nervous system. Studying neuroinflammation in vitro has been difficult because most primary culture models do not include all three critical cell types. We describe an in vitro model of neuroinflammation comprised of neurons, astrocytes and microglia. Primary rat cortical cells were cultured in a serum-free medium used to co-culture neurons and astrocytes that is supplemented with three factors (IL-34, TGF-β and cholesterol) used to support isolated microglia. This “tri-culture” can be maintained for at least 14 days in vitro while retaining a physiologically-relevant representation of all three cell types. Additionally, we demonstrate that the tri-culture system responds to lipopolysaccharide, mechanical trauma and excitotoxicity with both neurotoxic and neuroprotective aspects of the neuroinflammatory response observed in vivo. We expect the tri-culture model will enable mechanistic studies of neuroinflammation in vitro with enhanced physiological relevance.

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Senescent Phenotype of Astrocytes Leads to Microglia Activation and Neuronal Death

10.21203/rs.3.rs-1186816/v1 ◽

2021 ◽

Author(s):

You Wen Zhang ◽

Xuehan Yang ◽

Jingyue Liu ◽

Yichen Pan ◽

Ming Zhang ◽

...

Keyword(s):

Oxidative Stress ◽

Brain Aging ◽

Cell Model ◽

Neurological Diseases ◽

Ion Homeostasis ◽

Senescent Phenotype ◽

Age Related ◽

The Central Nervous System ◽

Aged Subjects

Abstract Astrocyte, the most abundant cell type in the central nervous system, is increasingly recognized and is thought to depend on curial and diverse roles in maintaining brain homeostasis, the blood-brain barrier, ion homeostasis, secrete neurotrophic factors and regulate synaptic transmission which is essential to tune individual-to-network neuronal activity. Senescence in astrocytes has been discovered to be an important contributor to several age-related neurological diseases like Alzheimer's and Parkinson's disease. However, the latest research about astrocytes from aged subjects or aged astrocytes in vitro is not yet adequate to be well elucidated on their curial process in the regulation of brain function. In this study, an in vitro cell model of aged astrocytes was constructed by serial passaging until passage 20-25, and those passages within 1-5 were used as young astrocytes. Meanwhile, oxidative induced astrocyte senescence model was also constructed by H2O2 induction. Our results indicate that after serial passaging or oxidative stress-induced astrocytes, all showed manifest changes in several established markers of cellular senescence like P53, P21, the release of inflammatory cytokine IL-6 and SA-β-gal positive cells. Results also showed mitochondrial dysfunction in the oxidative stress-induced astrocyte senescence model and treatment of berberine could reverse these alterations. What’s more interests us is that those two types of senescent astrocytes’ conditioned medium co-cultured with neuronal cells could do impact on neuron apoptosis no matter in direct or indirect ways. This study may help us better understand the fundamental role of astrocyte senescence on the regulation of normal and pathological brain aging.

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1120: Citrate and Vitamin E Blunt the SWL-Induced Free Radical Surge in an In-Vitro MDCK Cell Culture Model

The Journal of Urology ◽

10.1016/s0022-5347(18)38357-5 ◽

2004 ◽

Vol 171 (4S) ◽

pp. 295-295

Author(s):

Fernando C. Delvecchio ◽

Ricardo M. Brizuela ◽

Karen J. Byer ◽

W. Patrick Springhart ◽

Saeed R. Khan ◽

...

Keyword(s):

Cell Culture ◽

Free Radical ◽

Vitamin E ◽

Mdck Cell ◽

Cell Culture Model ◽

Culture Model

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Interferon-beta führt zu einer Stabilisierung der Barriereigenschaften in Hirnkapillarendothelzellen von vier verschiedenen Spezies in vitro

Aktuelle Neurologie ◽

10.1055/s-2005-919611 ◽

2005 ◽

Vol 32 (S 4) ◽

Author(s):

J Kraus ◽

K Voigt ◽

M Scholz ◽

A Schuller ◽

P Oschmann ◽

...

Keyword(s):

Interferon Beta

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Effect of [Asu,]eel calcitonin on prolactin release in normal subjects and patients with prolactinoma

Acta Endocrinologica ◽

10.1530/acta.0.1080297 ◽

1985 ◽

Vol 108 (3) ◽

pp. 297-304 ◽

Cited By ~ 6

Author(s):

Hidesuke Kaji ◽

Kazuo Chihara ◽

Naoto Minamitani ◽

Hitoshi Kodama ◽

Tetsuya Kita ◽

...

Keyword(s):

Body Weight ◽

Bolus Injection ◽

Normal Subjects ◽

Regular Insulin ◽

Prolactin Release ◽

Saline Administration ◽

The Central Nervous System ◽

Plasma Prl ◽

Male Subjects

Abstract. The effect of [Asu]eel calcitonin (ECT), an equipotent analogue of eel CT, on prolactin (Prl) secretion was examined in 12 healthy male subjects and in 6 patients with prolactinoma. In healthy subjects, ECT (0.5 μg/kg body weight · h) or saline was infused for 2 h and TRH was injected iv as a bolus of 500 μg at 1 h of ECT or saline administration. ECT did not affect basal Prl levels during 1 h of infusion. TRH caused a significant increase of plasma Prl with peak values of 75.2 ± 11.6 ng/ml in ECT-infused subjects, which did not differ from those infused with saline (68.5 ± 8.3 ng/ml). Next, an iv bolus injection of regular insulin (0.1 U/kg body weight) was followed by an infusion of ECT or saline alone. Plasma Prl peaks after hypoglycaemic stress were significantly lower in ECT-infused subjects than those in saline-injected controls (ECT, 16.5 ± 3.1 vs 33.5 ± 9.6 ng/ml, P < 0.05). In patients with prolactinoma, basal levels of plasma Prl ranging from 42.0–4130 ng/ml failed to change during iv infusion of ECT. Moreover, ECT (10−9–10−6m) did not affect Prl release from prolactinoma tissues perifused in vitro. These findings suggest that ECT may not act directly on the pituitary to modify Prl release. Rather, peripherally administered ECT appears to suppress Prl release via the central nervous system.

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Neural Stem Cells to Glial Cells an Important Factor for Brain Injury

Journal of Regenerative Biology and Medicine ◽

10.37191/mapsci-2582-385x-2(3)-025 ◽

2020 ◽

Author(s):

Prithiv K R Kumar

Keyword(s):

Stem Cells ◽

Central Nervous System ◽

Nervous System ◽

Neural Stem Cells ◽

Glial Cells ◽

Brain Injuries ◽

The Body ◽

The Central Nervous System ◽

Near Future

Stem cells have the capacity to differentiate into any type of cell or organ. Stems cell originate from any part of the body, including the brain. Brain cells or rather neural stem cells have the capacitive advantage of differentiating into the central nervous system leading to the formation of neurons and glial cells. Neural stem cells should have a source by editing DNA, or by mixings chemical enzymes of iPSCs. By this method, a limitless number of neuron stem cells can be obtained. Increase in supply of NSCs help in repairing glial cells which in-turn heal the central nervous system. Generally, brain injuries cause motor and sensory deficits leading to stroke. With all trials from novel therapeutic methods to enhanced rehabilitation time, the economy and quality of life is suppressed. Only PSCs have proven effective for grafting cells into NSCs. Neurons derived from stem cells is the only challenge that limits in-vitro usage in the near future.

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Beyond Oncological Hyperthermia: Physically Drivable Magnetic Nanobubbles as Novel Multipurpose Theranostic Carriers in the Central Nervous System

Molecules ◽

10.3390/molecules25092104 ◽

2020 ◽

Vol 25 (9) ◽

pp. 2104 ◽

Cited By ~ 1

Author(s):

Eleonora Ficiarà ◽

Shoeb Anwar Ansari ◽

Monica Argenziano ◽

Luigi Cangemi ◽

Chiara Monge ◽

...

Keyword(s):

Central Nervous System ◽

Nervous System ◽

Brain Tumors ◽

Ad Hoc ◽

Superparamagnetic Iron Oxide ◽

Conventional Radiotherapy ◽

The Central Nervous System ◽

Magnetic Resonance Imaging Mri ◽

The Brain

Magnetic Oxygen-Loaded Nanobubbles (MOLNBs), manufactured by adding Superparamagnetic Iron Oxide Nanoparticles (SPIONs) on the surface of polymeric nanobubbles, are investigated as theranostic carriers for delivering oxygen and chemotherapy to brain tumors. Physicochemical and cyto-toxicological properties and in vitro internalization by human brain microvascular endothelial cells as well as the motion of MOLNBs in a static magnetic field were investigated. MOLNBs are safe oxygen-loaded vectors able to overcome the brain membranes and drivable through the Central Nervous System (CNS) to deliver their cargoes to specific sites of interest. In addition, MOLNBs are monitorable either via Magnetic Resonance Imaging (MRI) or Ultrasound (US) sonography. MOLNBs can find application in targeting brain tumors since they can enhance conventional radiotherapy and deliver chemotherapy being driven by ad hoc tailored magnetic fields under MRI and/or US monitoring.

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An In Vitro Cell Culture Model for Pyoverdine-Mediated Virulence

Pathogens ◽

10.3390/pathogens10010009 ◽

2020 ◽

Vol 10 (1) ◽

pp. 9

Author(s):

Donghoon Kang ◽

Natalia V. Kirienko

Keyword(s):

Cell Culture ◽

Virulence Factors ◽

Model Organisms ◽

Cell Culture Model ◽

Chronic Infections ◽

In Vitro Cell Culture ◽

Mitochondrial Homeostasis ◽

Culture Model ◽

Wide Range

Pseudomonas aeruginosa is a multidrug-resistant, opportunistic pathogen that utilizes a wide-range of virulence factors to cause acute, life-threatening infections in immunocompromised patients, especially those in intensive care units. It also causes debilitating chronic infections that shorten lives and worsen the quality of life for cystic fibrosis patients. One of the key virulence factors in P. aeruginosa is the siderophore pyoverdine, which provides the pathogen with iron during infection, regulates the production of secreted toxins, and disrupts host iron and mitochondrial homeostasis. These roles have been characterized in model organisms such as Caenorhabditis elegans and mice. However, an intermediary system, using cell culture to investigate the activity of this siderophore has been absent. In this report, we describe such a system, using murine macrophages treated with pyoverdine. We demonstrate that pyoverdine-rich filtrates from P. aeruginosa exhibit substantial cytotoxicity, and that the inhibition of pyoverdine production (genetic or chemical) is sufficient to mitigate virulence. Furthermore, consistent with previous observations made in C. elegans, pyoverdine translocates into cells and disrupts host mitochondrial homeostasis. Most importantly, we observe a strong correlation between pyoverdine production and virulence in P. aeruginosa clinical isolates, confirming pyoverdine’s value as a promising target for therapeutic intervention. This in vitro cell culture model will allow rapid validation of pyoverdine antivirulents in a simple but physiologically relevant manner.

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Miniaturization and Automation of a Human In Vitro Blood–Brain Barrier Model for the High-Throughput Screening of Compounds in the Early Stage of Drug Discovery

Pharmaceutics ◽

10.3390/pharmaceutics13060892 ◽

2021 ◽

Vol 13 (6) ◽

pp. 892

Author(s):

Elisa L. J. Moya ◽

Elodie Vandenhaute ◽

Eleonora Rizzi ◽

Marie-Christine Boucau ◽

Johan Hachani ◽

...

Keyword(s):

Drug Discovery ◽

Blood Brain Barrier ◽

Early Stage ◽

Molecular Transport ◽

Brain Barrier ◽

Blood Brain ◽

Cns Drugs ◽

The Impact ◽

The Brain

Central nervous system (CNS) diseases are one of the top causes of death worldwide. As there is a difficulty of drug penetration into the brain due to the blood–brain barrier (BBB), many CNS drugs treatments fail in clinical trials. Hence, there is a need to develop effective CNS drugs following strategies for delivery to the brain by better selecting them as early as possible during the drug discovery process. The use of in vitro BBB models has proved useful to evaluate the impact of drugs/compounds toxicity, BBB permeation rates and molecular transport mechanisms within the brain cells in academic research and early-stage drug discovery. However, these studies that require biological material (animal brain or human cells) are time-consuming and involve costly amounts of materials and plastic wastes due to the format of the models. Hence, to adapt to the high yields needed in early-stage drug discoveries for compound screenings, a patented well-established human in vitro BBB model was miniaturized and automated into a 96-well format. This replicate met all the BBB model reliability criteria to get predictive results, allowing a significant reduction in biological materials, waste and a higher screening capacity for being extensively used during early-stage drug discovery studies.

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