scholarly journals Abnormal Cannabidiol Affects Production of Pro-Inflammatory Mediators and Astrocyte Wound Closure in Primary Astrocytic-Microglial Cocultures

Molecules ◽  
2020 ◽  
Vol 25 (3) ◽  
pp. 496 ◽  
Author(s):  
Julian Cardinal von Widdern ◽  
Tim Hohmann ◽  
Faramarz Dehghani
Keyword(s):  
Knockout Mice ◽  
Wound Closure ◽  
Scar Formation ◽  
Neuroprotective Effects ◽  
Proinflammatory Mediators ◽  
Nitrite Production ◽  
Scratch Wound ◽  
Scratch Wound Assay ◽  
The Impact

Abnormal cannabidiol (abn-CBD) exerts neuroprotective effects in vivo and in vitro. In the present study, we investigated the impact of abn-CBD on the glial production of proinflammatory mediators and scar formation within in vitro models. Primary astrocytic-microglial cocultures and astrocytic cultures from neonatal C57BL/6 mice and CB2 receptor knockout mice were stimulated with lipopolysaccharide (LPS), and the concentrations of tumor necrosis factor α (TNFα), interleukin-6 (IL-6) and nitrite were determined. Furthermore, we performed a live cell microscopy-based scratch-wound assay. After LPS stimulation, TNFα, IL-6 and nitrite production was more strongly increased in cocultures than in isolated astrocytes. Abn-CBD treatment attenuated the LPS-induced production of TNFα and nitrite in cocultures, while IL-6 production remained unaltered. In isolated astrocytes, only LPS-induced TNFα production was reduced by abn-CBD. Similar effects were observed after abn-CBD application in cocultures of CB2 knockout mice. Interestingly, LPS-induced TNFα and nitrite levels were far lower in CB2 knockout cultures compared to wildtypes, while IL-6 levels did not differ. In the scratch-wound assay, treatment with abn-CBD decelerated wound closure when microglial cells were present. Our data shows a differential role of abn-CBD for modulation of glial inflammation and astrocytic scar formation. These findings provide new explanations for mechanisms behind the neuroprotective potential of abn-CBD.

Auswirkungen Konservierungsmittel-freier Fluorchinolon-Augenlösungen auf menschliche Hornhaut-Epithelzellen in vitro

2015 ◽  
Vol 1 (1) ◽  
pp. 26-27
Keyword(s):  
Scratch Wound ◽  
Scratch Wound Assay

Ziele: Die Beurteilung der biologischen Wirkungen Konservierungsmittel-freier Fluorchinolon-Augenlösungen auf Zellkulturen menschlichen Hornhaut-Epithels in vitro.Methoden: Wir untersuchten die Auswirkung von topischen Fluorchinolonen verschiedener Generationen, wie Ofloxacin 0,3%, Levofloxacin 0,5%, Tosufloxacin 0,3%, Moxifloxacin 0,5% und Gatifloxacin 0,3%, auf gezüchtete menschliche Hornhaut-Epithelzellen. Die Untersuchung erfolgte mittels MTT-basiertem kalorimetrischem Assay, Laktatdehydrogenase(LDH)-Leakage-Assay und Scratch-Wound-Assay. Die Morphologie der Hornhaut-Epithelzellen wurde mittels inverser Lichtmikroskopie und Transmissionselektronenmikroskopie untersucht.Ergebnisse: Bei allen topischen Fluorchinolonen ging die metabolische Aktivität der Hornhaut-Epithelzellen zeitabhängig zurück, und der LDH-Titer stieg mit zunehmender Dauer der Wirkstoffexposition. Insbesondere nach einer Exposition gegenüber Moxifloxacin 0,5% und Gatifloxacin 0,3% war ein signifikanter Anstieg der LDH-Titer im Vergleich zu den Kontrollen festzustellen. Die Migrationsraten der Hornhaut-Epithelzellen waren bei Ofloxacin 0,3% und Levofloxacin 0,5% höher als bei den anderen Fluorchinolonen. Nach einer Exposition gegenüber Moxifloxacin 0,5% und Gatifloxacin 0,3% waren schwere morphologische Schäden an den Zellen zu beobachten.Schlussfolgerung: Da Moxifloxacin 0,5% und Gatifloxacin 0,3% eine stärkere toxische Wirkung auf die Hornhaut-Epithelzellen ausübten als die anderen Fluorchinolone, sind diese Fluorchinolon-Augenlösungen der 4. Generation nur nach sorgfältiger Abwägung im Hinblick auf die mögliche Schädigung des Hornhaut-Epithels bei langer Behandlungsdauer oder zu hoher Dosierung anzuwenden.Übersetzung aus Ophthalmic Res 2014;51:216-223 (DOI: 10.1159/000357976)

scholarly journals Sigma 1 Receptor is Overexpressed in Hepatocellular Adenoma: Involvement of ERα and HNF1α

Cancers ◽  
2020 ◽  
Vol 12 (8) ◽  
pp. 2213
Author(s):  
Laure Villemain ◽  
Sylvie Prigent ◽  
Aurélie Abou-Lovergne ◽  
Laura Pelletier ◽  
Magali Chiral ◽  
...  
Keyword(s):  
Knockout Mice ◽  
Hepatocellular Adenoma ◽  
Integral Membrane Protein ◽  
Hepatocyte Proliferation ◽  
Sigma 1 Receptor ◽  
Cellular Models ◽  
Huh7 Cells ◽  
The Impact

Sigma receptor 1 (SigR1) is an endoplasmic reticulum resident integral membrane protein whose functions remain unclear. Although the liver shows the highest expression of SigR1, its role in this organ is unknown. SigR1 is overexpressed in many cancers and its expression is correlated to hormonal status in hormone-dependent cancers. To better understand the role of SigR1 in hepatocytes we focused our work on the regulation of its expression in tumoral liver. In this context, hepatocellular adenomas, benign hepatic tumors associated with estrogen intake are of particular interest. The expression of SigR1 mRNA was assessed in hepatocellular adenoma (HCA) patients using qPCR. The impact of estrogen on the expression of SigR1 was studied in vivo (mice) and in vitro (HepG2 and Huh7 cells). The effect of HNF1α on the expression of SigR1 was studied in vivo by comparing wild type mice to HNF1 knockout mice. Estrogen enhanced SigR1 expression through its nuclear receptor ERα. HNF1α mutated HCA (H-HCA) significantly overexpressed SigR1 compared to all other HCA subtypes. HNF1 knockout mice showed an increase in SigR1 expression. Overexpressing SigR1 in cellular models increases proliferation rate and storage of lipid droplets, which phenocopies the H-HCA phenotype. SigR1 is involved in hepatocyte proliferation and steatosis and may play an important role in the control of the H-HCA phenotype.

scholarly journals Impact of Kefir DerivedLactobacillus kefirion the Mucosal Immune Response and Gut Microbiota

2015 ◽  
Vol 2015 ◽  
pp. 1-12 ◽  
Author(s):  
P. Carasi ◽  
S. M. Racedo ◽  
C. Jacquot ◽  
D. E. Romanin ◽  
M. A. Serradell ◽  
...  
Keyword(s):  
Fecal Microbiota ◽  
Mucosal Immune Response ◽  
Pcr Analysis ◽  
Mesenteric Lymph Nodes ◽  
Reporter System ◽  
Gm Csf ◽  
Proinflammatory Mediators ◽  
Probiotic Treatment ◽  
The Impact

The evaluation of the impact of probiotics on host health could help to understand how they can be used in the prevention of diseases. On the basis of our previous studies andin vitroassays on PBMC and Caco-2 ccl20:luc reporter system presented in this work, the strainLactobacillus kefiriCIDCA 8348 was selected and administrated to healthy Swiss mice daily for 21 days. The probiotic treatment increased IgA in feces and reduced expression of proinflammatory mediators in Peyer Patches and mesenteric lymph nodes, where it also increased IL-10. In ileum IL-10, CXCL-1 and mucin 6 genes were upregulated; meanwhile in colon mucin 4 was induced whereas IFN-γ, GM-CSF, and IL-1βgenes were downregulated. Moreover, ileum and colon explants showed the anti-inflammatory effect ofL. kefirisince the LPS-induced increment of IL-6 and GM-CSF levels in control mice was significantly attenuated inL. kefiritreated mice. Regarding fecal microbiota, DGGE profiles allowed differentiation of experimental groups in two separated clusters. Quantitative PCR analysis of different bacterial groups revealed only significant changes inLactobacilluspopulation. In conclusion,L. kefiriis a good candidate to be used in gut inflammatory disorders.

scholarly journals Beneficial Effect of Tempol, a Membrane-Permeable Radical Scavenger, on Inflammation and Osteoarthritis in In Vitro Models

Biomolecules ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 352
Author(s):  
Giovanna Calabrese ◽  
Alessio Ardizzone ◽  
Michela Campolo ◽  
Sabrina Conoci ◽  
Emanuela Esposito ◽  
...  
Keyword(s):  
Biological Properties ◽  
In Vitro Models ◽  
Radical Scavenger ◽  
Proinflammatory Mediators ◽  
Nitrite Production ◽  
Inflammatory State ◽  
Anti Inflammatory ◽  
Vitro Model ◽  
Antioxidant Effects

Osteoarthritis (OA) is one of the most common and widespread diseases which is highly disabling for humans. This makes OA a chronic disease for which it is urgent to find new therapeutic strategies. The inflammatory state in OA contributes to its progression through multiple mechanisms involving the recruitment of phagocytes and leukocytes, inflammatory response, and reactive oxygen species (ROS) production. Tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl) is classifiable as a piperidine nitroxide, with excellent antioxidant effects, while its anti-inflammatory role is not yet clear. On this basis, we explored its promising biological properties in two in vitro model:, macrophage (J774) and chondrocyte (CC) cell lines. With this aim in mind, we induced inflammation in J774 and CC using lipopolysaccharide (LPS) and Interleukin1β (IL-1β), and after 24, 72 and 168 h of tempol treatment analyzed their effects on cytotoxicity and anti-inflammatory activity. Our data suggested that tempol treatment is able to reduce inflammation and nitrite production in LPS-induced J774 as well as reducing the production of proinflammatory mediators including cytokines, enzymes, and metalloproteases (MMPs) in IL-1β-stimulated CC. Thus, since inflammation and oxidative stress have a crucial role in the pathogenesis and progression of OA, tempol could be considered as a new therapeutic approach for this pathology.

Delayed skin wound repair in proline-rich protein tyrosine kinase 2 knockout mice

2014 ◽  
Vol 306 (10) ◽  
pp. C899-C909 ◽  
Author(s):  
Aaron C. Koppel ◽  
Alexi Kiss ◽  
Anna Hindes ◽  
Carole J. Burns ◽  
Barry L. Marmer ◽  
...  
Keyword(s):  
Wound Repair ◽  
Protein Tyrosine Kinase ◽  
Wound Closure ◽  
Skin Wound ◽  
Dominant Negative ◽  
Dominant Negative Mutant ◽  
Tyrosine Kinase 2 ◽  
Scratch Wound ◽  
Protein Tyrosine Kinase 2

Proline-rich protein tyrosine kinase 2 (Pyk2) is a member of the focal adhesion kinase family. We used Pyk2 knockout (Pyk2-KO) mice to study the role of Pyk2 in cutaneous wound repair. We report that the rate of wound closure was delayed in Pyk2-KO compared with control mice. To examine whether impaired wound healing of Pyk2-KO mice was caused by a keratinocyte cell-autonomous defect, the capacities of primary keratinocytes from Pyk2-KO and wild-type (WT) littermates to heal scratch wounds in vitro were compared. The rate of scratch wound repair was decreased in Pyk2-KO keratinocytes compared with WT cells. Moreover, cultured human epidermal keratinocytes overexpressing the dominant-negative mutant of Pyk2 failed to heal scratch wounds. Conversely, stimulation of Pyk2-dependent signaling via WT Pyk2 overexpression induced accelerated scratch wound closure and was associated with increased expression of matrix metalloproteinase (MMP)-1, MMP-9, and MMP-10. The Pyk2-stimulated increase in the rate of scratch wound repair was abolished by coexpression of the dominant-negative mutant of PKCδ and by GM-6001, a broad-spectrum inhibitor of MMP activity. These results suggest that Pyk2 is essential for skin wound reepithelialization in vivo and in vitro and that it regulates epidermal keratinocyte migration via a pathway that requires PKCδ and MMP functions.

scholarly journals Characterization of genetically modified mice for phosphoglycerate mutase, a vitally-essential enzyme in glycolysis

PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0250856
Author(s):  
Takumi Mikawa ◽  
Eri Shibata ◽  
Midori Shimada ◽  
Ken Ito ◽  
Tomiko Ito ◽  
...  
Keyword(s):  
Glucose Metabolism ◽  
Knockout Mice ◽  
Physiological Role ◽  
Phosphoglycerate Mutase ◽  
Physiological Homeostasis ◽  
Essential Enzyme ◽  
The Impact

Glycolytic metabolism is closely involved in physiological homeostasis and pathophysiological states. Among glycolytic enzymes, phosphoglycerate mutase (PGAM) has been reported to exert certain physiological role in vitro, whereas its impact on glucose metabolism in vivo remains unclear. Here, we report the characterization of Pgam1 knockout mice. We observed that homozygous knockout mice of Pgam1 were embryonic lethal. Although we previously reported that both PGAM-1 and -2 affect global glycolytic profile of cancers in vitro, in vivo glucose parameters were less affected both in the heterozygous knockout of Pgam1 and in Pgam2 transgenic mice. Thus, the impact of PGAM on in vivo glucose metabolism is rather complex than expected before.

Effect of different chitosan derivatives on in vitro scratch wound assay: A comparative study

2015 ◽  
Vol 76 ◽  
pp. 236-241 ◽  
Author(s):  
Francesca Felice ◽  
Ylenia Zambito ◽  
Ester Belardinelli ◽  
Angela Fabiano ◽  
Tatiana Santoni ◽  
...  
Keyword(s):  
Comparative Study ◽  
Chitosan Derivatives ◽  
Scratch Wound ◽  
Scratch Wound Assay

scholarly journals The Protective Role of E-64d in Hippocampal Excitotoxic Neuronal Injury Induced by Glutamate in HT22 Hippocampal Neuronal Cells

2021 ◽  
Vol 2021 ◽  
pp. 1-14
Author(s):  
RuiJin Xie ◽  
TianXiao Li ◽  
XinYu Qiao ◽  
HuiYa Mei ◽  
GuoQin Hu ◽  
...  
Keyword(s):  
Neuroprotective Effect ◽  
Epileptic Seizures ◽  
Mitochondrial Pathway ◽  
Neuroprotective Effects ◽  
Ht22 Cells ◽  
Pathway Activity ◽  
Metabolism Enzymes ◽  
The Impact ◽  
Chaperone Mediated Autophagy

Epilepsy is the most common childhood neurologic disorder. Status epilepticus (SE), which refers to continuous epileptic seizures, occurs more frequently in children than in adults, and approximately 40–50% of all cases occur in children under 2 years of age. Conventional antiepileptic drugs currently used in clinical practice have a number of adverse side effects. Drug-resistant epilepsy (DRE) can progressively develop in children with persistent SE, necessitating the development of novel therapeutic drugs. During SE, the persistent activation of neurons leads to decreased glutamate clearance with corresponding glutamate accumulation in the synaptic extracellular space, increasing the chance of neuronal excitotoxicity. Our previous study demonstrated that after developmental seizures in rats, E-64d exerts a neuroprotective effect on the seizure-induced brain damage by modulating lipid metabolism enzymes, especially ApoE and ApoJ/clusterin. In this study, we investigated the impact and mechanisms of E-64d administration on neuronal excitotoxicity. To test our hypothesis that E-64d confers neuroprotective effects by regulating autophagy and mitochondrial pathway activity, we simulated neuronal excitotoxicity in vitro using an immortalized hippocampal neuron cell line (HT22). We found that E-64d improved cell viability while reducing oxidative stress and neuronal apoptosis. In addition, E-64d treatment regulated mitochondrial pathway activity and inhibited chaperone-mediated autophagy in HT22 cells. Our findings indicate that E-64d may alleviate glutamate-induced damage via regulation of mitochondrial fission and apoptosis, as well as inhibition of chaperone-mediated autophagy. Thus, E-64d may be a promising therapeutic treatment for hippocampal injury associated with SE.

Synthesis and evaluation of an alginate-methacrylate xerogel for insulin delivery towards wound healing applications

2021 ◽  
Vol 12 (3) ◽  
pp. 215-234
Author(s):  
GP Rajalekshmy ◽  
MR Rekha
Keyword(s):  
Wound Healing ◽  
Transmission Rate ◽  
Wound Care ◽  
Physical Stability ◽  
Analytical Techniques ◽  
Insulin Delivery ◽  
Water Vapour Transmission Rate ◽  
Scratch Wound ◽  
Scratch Wound Assay

Background: Alginate is one of the most widely used biopolymer for wound healing. But poor mechanical strength and degradability limits its application especially as a drug-delivery matrix. The aim of this study was to develop stable alginate based scaffold for insulin delivery toward wound care. Materials & methods: The xerogel alginate-g-poly (methacrylic acid; AGM2S) was characterized by various analytical techniques. Results: AGM2S xerogel showed improved physical stability, low degradation, good swelling and water vapour transmission rate (WVTR). About 70% of insulin was released from loaded xerogel over a period of 48 h and favorably modulated the healing response in in vitro scratch wound assay. Conclusion: Grafting improved the strength and stability of alginate xerogel and the results suggest the application of insulin loaded AGM2S xerogels as a potential wound healing material.

scholarly journals Avaliação dos efeitos do inibidor multiquinase sorafenibe sobre as células musculares lisas arteriais de Rattus norvegicus: Estudo in vitro / Evaluation of the effects of the multikinase inhibitor sorafenib on the arterial smooth muscle cells of Rattus norvegicus: An in vitro study

2021 ◽  
Vol 7 (12) ◽  
pp. 116307-116318
Author(s):  
Rafael de Nogueira Riberio ◽  
Gabriela Mendonça Dos Reis ◽  
Laura Bainy Rodrigues De Freitas ◽  
Gabriela Jouglard Vasquez Amado ◽  
Jessica Machado Miri ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Rattus Norvegicus ◽  
In Vitro Study ◽  
Multikinase Inhibitor ◽  
Scratch Wound ◽  
Scratch Wound Assay ◽  
Ciclo Celular ◽  
Arterial Smooth Muscle Cells

A reestenose arterial é um processo inflamatório que pode ocorrer após colocação de stent por cateterismo. Os stents farmacológicos surgiram para reduzir esse problema e o inibidor multiquinase sorafenibe demonstrou ser um composto com ação efetiva. Este estudo in vitro avaliou os efeitos do sorafenibe sobre a citotoxicidade, migração celular e distribuição das células nas fases do ciclo celular. A linhagem celular de músculo liso de rato A7r5 foi tratada com sorafenibe em concentrações que variaram de 0 a 5 μM. Os efeitos citotóxicos foram avaliados por dois ensaios colorimétricos, MTT e SRB após 24 horas de tratamento. A distribuição das células nas fases do ciclo celular foi avaliada por citometria de fluxo e a capacidade de cicatrização/migração celular pelo ensaio scratch wound assay. Comparado com o controle positivo paclitaxel, o sorafenibe demonstrou um efeito 1,6 vezes maior na redução da proliferação celular. Na avaliação do ciclo celular, o sorafenibe mostrou um bloqueio na fase G0/G1. Além disso, o sorafenibe aumentou o número de A7r5 células na fase sub-G1, sugerindo morte celular. No entanto, no estudo de cicatrização/migração celular, não foi observado efeito quando comparado ao controle negativo. Assim, esses resultados in vitro sugerem que o sorafenibe é eficaz para uso em stents farmacológicos, sugerindo uma continuidade na investigação desse fármaco.

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