C6 Glioma Cell Line Research Articles

Guar gum and carboxymethyl cellulose nanocomposite loaded with piperine: A multipurpose nanoparticle with potential antibacterial and anticancer properties

Guar gum-carboxymethyl cellulose derived piperine nanocomposite (GG-CMC@PIP) was synthesized to develop a phytochemical-based nanomaterial by following a simple synthetic procedure. In this study, piperine, an alkaloid prodrug was nanonized by entrapping within guar gum-carboxymethyl cellulose derived nanocomposite (GG-CMC). GG-CMC@PIPs were of (i) size (25 ± 3) nm by TEM, (ii) zeta potential (−) 33.1 ± 0.73 mV and (iii) entrapment efficiency 86 ± 0.46 %. High loading percentage of piperine in polymeric nanocomposites revealed excellent incorporation efficiency. In addition, pH responsive release behaviour of piperine from nanocomposites exhibited sustained but faster release pattern in pH 5.5. Minimal inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of piperine loaded nanocomposite on Gram positive Enterococcus faecalis and Gram-negative Escherichia coli, Pseudomonas aeruginosa were in the range of 200–400 μg/ml. IC50 value of GG-CMC@PIP was found to be around 500 μg/ml against C6 glioma cell line. Experimental outcome confirmed excessive accumulation of reactive oxygen species (ROS) mediated cytotoxicity, mitochondrial depolarization and higher degree of nuclear damage followed by increased mortality of C6 glioma cell after treatment with piperine-loaded nanocomposite.

Design and synthesis of new benzothiazole-piperazine derivatives and in vitro and in silico investigation of their anticancer activity

In this work, novel benzothiazole-piperazine acetamide (2a-2l) analogs were synthesized and evaluated their anticancer activity. The 1HNMR , 13CNMR and LC-MS/MS spectral data and elemental analyses were used to clarify the structure of the final molecules. The title compounds were procured by reacting 2‑chloro-N-(6-substituted benzothiazole-2-yl) acetamide with some piperazine derivatives. Cytotoxicity and apoptosis parameters including Annexin V binding capacities, effect on cell cycle, caspase-3 activation and mitochondrial membrane depolarization effect were evaluated to determine anticancer profile of the final molecules on A549 (human lung adenocarcinoma), C6 (glioma cell line) and NIH/3T3 (mouse embryoblast cell line). Mostly, the final molecules showed significant cytotoxic profile with prominent selectivity. 2c (IC50: 7.23±1.17, SI: 6.93), 2d (IC50: 17.50±4.95; SI: 11.59), and 2 h (IC50: 10.83±0.76; SI: 29.23) showed significant and selective cytotoxic activity. Compounds 2b, 2c and 2e provoked apoptosis of A549 cells with potential higher than cisplatin as determined by flow cytometry. The cell cycle analysis of 2b, 2e, 2 g, and 2 h suggested that the A549 cells were affected during G0/G1 phase. Caspase-3 activation was observed on cells by compounds 2a, 2e, 2i and 2l at most. The highest mitochondrial membrane depolarization ratios were seen treated by compounds 2e, 2 h, and 2j. None of the analogs displayed any obvious inhibition activity on MMP-9. The physicochemical properties were predicted for all compounds and also molecular docking and dynamics simulation studies were realized for compound 2e, namely N-(6-methoxybenzothiazol-2-yl)-2-(4-(p-tolyl) piperazin-1-yl) acetamide. The results indicated that merging benzothiazole and piperazine derivatives via acetamide bridge is promising in cancer research because these moieties can interact with the loop and ß-sheet regions of MMP-9. So, 2e was considered as a potential therapeutic agent against lung carcinoma.

Effect of Xanthohumol, a Bioactive Natural Compound from Hops, on Adenosine Pathway in Rat C6 Glioma and Human SH-SY5Y Neuroblastoma Cell Lines.

Xanthohumol (Xn) is an antioxidant flavonoid mainly extracted from hops (Humulus lupulus), one of the main ingredients of beer. As with other bioactive compounds, their therapeutic potential against different diseases has been tested, one of which is Alzheimer's disease (AD). Adenosine is a neuromodulatory nucleoside that acts through four different G protein-coupled receptors: A1 and A3, which inhibit the adenylyl cyclases (AC) pathway, and A2A and A2B, which stimulate this activity, causing either a decrease or an increase, respectively, in the release of excitatory neurotransmitters such as glutamate. This adenosinergic pathway, which is altered in AD, could be involved in the excitotoxicity process. Therefore, the aim of this work is to describe the effect of Xn on the adenosinergic pathway using cell lines. For this purpose, two different cellular models, rat glioma C6 and human neuroblastoma SH-SY5Y, were exposed to a non-cytotoxic 10 µM Xn concentration. Adenosine A1 and A2A, receptor levels, and activities related to the adenosine pathway, such as adenylate cyclase, protein kinase A, and 5'-nucleotidase, were analyzed. The adenosine A1 receptor was significantly increased after Xn exposure, while no changes in A2A receptor membrane levels or AC activity were reported. Regarding 5'-nucleotidases, modulation of their activity by Xn was noted since CD73, the extracellular membrane attached to 5'-nucleotidase, was significantly decreased in the C6 cell line. In conclusion, here we describe a novel pathway in which the bioactive flavonoid Xn could have potentially beneficial effects on AD as it increases membrane A1 receptors while modulating enzymes related to the adenosine pathway in cell cultures.

In Vitro and In Silico Anti-Glioblastoma Activity of Hydroalcoholic Extracts of Artemisia annua L. and Artemisia vulgaris L.

Glioblastoma, the most aggressive and challenging brain tumor, is a key focus in neuro-oncology due to its rapid growth and poor prognosis. The C6 glioma cell line is often used as a glioblastoma model due to its close simulation of human glioma characteristics, including rapid expansion and invasiveness. Alongside, herbal medicine, particularly Artemisia spp., is gaining attention for its anticancer potential, offering mechanisms like apoptosis induction, cell cycle arrest, and the inhibition of angiogenesis. In this study, we optimized extraction conditions of polyphenols from Artemisia annua L. and Artemisia vulgaris L. herbs and investigated their anticancer effects in silico and in vitro. Molecular docking of the main phenolic compounds of A. annua and A. vulgaris and potential target proteins, including programmed cell death (apoptosis) pathway proteins proapoptotic Bax (PDB ID 6EB6), anti-apoptotic Bcl-2 (PDB ID G5M), and the necroptosis pathway protein (PDB ID 7MON), mixed lineage kinase domain-like protein (MLKL), in complex with receptor-interacting serine/threonine-protein kinase 3 (RIPK3), revealed the high probability of their interactions, highlighting the possible influence of chlorogenic acid in modulating necroptosis processes. The cell viability of rat C6 glioma cell line was assessed using a nuclear fluorescent double-staining assay with Hoechst 33342 and propidium iodide. The extracts from A. annua and A. vulgaris have demonstrated anticancer activity in the glioblastoma model, with the synergistic effects of their combined compounds surpassing the efficacy of any single compound. Our results suggest the potential of these extracts as a basis for developing more effective glioblastoma treatments, emphasizing the importance of further research into their mechanisms of action and therapeutic applications.

Comparison of apoptotic effects of lupeol on A549 and C6 cell lines

Lupeol compound is a cyclic triterpene alcohol that is widely found in plants. The compound lupeol has been reported to exhibit antitumor, anti-inflammatory, anxiolytic, neuroprotective and hepatoprotective effects. Recent research shows that lupeol could be a potential medicine for various diseases and also an adjuvant for intractable diseases. Cancer poses a health threat that is increasingly common around the world. Among new cancer cases, lung cancer is one of the most common and deadly cancers worldwide. In this study, the anticancer efficiency of lupeol on human lung adenocarcinoma A549 and glioma C6 cell lines were examined. Various concentrations of lupeol (500, 250, 125, 62.5, 31.25, 15.62, 7.81, 3.90 μM) effectually reduced cell viability in the A549 and C6 cell lines in a dose-dependent manner. These results showed that lupeol had selective anticancer activity against A549 and C6 cell lines.

Synthesis, self-aggregation, thermal stability and cytotoxicity trends of alkyloxypropoxycyclohexane based pyridinium ionic liquids

New ether-functionalized pyridinium ionic liquids (ILs) has been synthesized by simple methodology and described by different spectroscopic techniques. Presence of two positional isomers has been confirmed with the help of NMR spectroscopy. The critical micelle concentration and other surface parameters of these ILs were deduced by conductivity, surface tension and fluorescence method. A collection of useful physical parameters i.e. critical micelle concentration (cmc), surface tension accomplished at cmc (γcmc), effectiveness of diminution in surface tension (Πcmc), maximum surface excess concentration (Ʈmax) and minimum surface area per molecule (Amin) at the air–water interface were also estimated. The average size of aggregates formed by investigated pyridinium ILs have been well examined by dynamic light scattering (DLS) experiments. The thermal degradation analysis of the present ILs has also been suggested that these ILs possessed thermal stability ranges from 200 °C to 240 °C. The investigated ILs have also been tested by MTT assay (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) on the C6 glioma cell line and found to revealed very low cytotoxicity in the concentration range required for biomedical applications. Such a low cytotoxicity of present ILs opens several new applications in the many biological purposes like drug and gene delivery agents. The physical study of investigated ILs indicate that the presence of ether functionality and varying hydrophobic chain length significantly altered the physicochemical properties of these pyridinium ILs than related to earlier synthesized N-(cyclohexanoxycarbonylmethyl) and 3-Cyclohexyloxy-2-Hydroxypropyl Pyridinium and Imidazolium surface active ILs.

Anoikis and cancer cell differentiation: novel modes of shikonin derivatives anticancer action in vitro.

Shikonin is a naturally occurring naphthoquinone found in the roots of several genera of the Boraginaceae family, widely known for its numerous biological activities, such as antiinflammatory, antioxidant, antimicrobial and anticancer. In this study, the antitumor effect of six naphthoquinones isolated from the roots of Onosma visianii was evaluated using two cell lines, mouse melanoma B16 and highly aggressive rat glioma cell line C6. All examined shikonins dose-dependently decreased the viability of tested cells, with compounds 5 and 6 being the most potent ones and hence subjected to further analysis. The diminished viability of B16 melanoma cells was in correlation with detected caspase-mediated apoptosis. Importantly, observed altered cell morphology along with the loss of dividing potential upon exposure to both shikonins implied reprogram of B16 cell phenotype. Elevated expression of myelin basic protein indicated the acquirement of Schwann-like cell phenotype, while detected autophagy might be connected to this phenomenon. On the contrary, upon exposure to both agents, C6 cells underwent specific cell death-anoikis, provoked by detachment from the extracellular matrix and compromised integrin signaling. Oppositely to compound 5, compound 6 realized anoikis in a caspase-independent manner and under sustained ERK1/2 activation, indicating the deviation from standard proanoikis signaling. Herein, we have pointed out the diversity and novelty in the mode of action of shikonin derivatives depending on the tumor cell features, which represents a good platform for new investigations of these promising natural compounds.

Borneol promotes autophagic degradation of HIF-1α and enhances chemotherapy sensitivity in malignant glioma.

Gliomas are characterized by high mortality rates and resistance. Even with conventional chemotherapy the prognosis of glioblastoma remains poor. Many medications are not optimally effective due to limited bioavailability. The bioavailability of medicine can be enhanced by borneol, a monoterpenoid substance. In this study, we investigated the effect of borneol, a commonly used Chinese medicine, on chemosensitivity in C6 glioma and U251 human glioma cell lines and elucidated its therapeutic molecular targets. The chemosensitivity-inducing effects of borneol in C6 and U251 cells were examined using CCK8 and clonal formation assays. The mechanism underlying the effect of borneol was evaluated through immunohistochemistry and western blotting assays. Further, the number of autophagosomes was determined via transmission electron microscopy. Finally, the chemical sensitization effect of borneol was evaluated in SD rats after C6 orthotopic tumor transplantation. Borneol increased cytotoxicity in C6 and U251 cells in response to temozolomide (TMZ). In addition, through transmission electron microscopy, western blotting, and immunohistochemical tests, we found that borneol combined with TMZ significantly increased the level of autophagy and that hypoxia inducible factor-1(HIF-1α) is a candidate target through which borneol enhances the cytotoxic effect of TMZ. Borneol's ability to enhance HIF-1α degradation was counteracted following the administration of autophagy inhibitors. In vivo, borneol treatment was found to enhance the anticancer effect of TMZ and delay tumor progression, and this effect was closely related to its ability to promote the autophagic degradation of HIF-1α. HIF-1α might be a valid therapeutic target of borneol, which can be potentially applied as a chemosensitizing drug used for glioma treatment.

CLEC19A overexpression inhibits tumor cell proliferation/migration and promotes apoptosis concomitant suppression of PI3K/AKT/NF-κB signaling pathway in glioblastoma multiforme.

GBM is the most frequent malignant primary brain tumor in humans. The CLEC19A is a member of the C-type lectin family, which has a high expression in brain tissue. Herein, we sought to carry out an in-depth analysis to pinpoint the role of CLEC19A expression in GBM. To determine the localization of CLEC19A, this protein was detected using Western blot, Immunocytochemistry/Immunofluorescence, and confocal microscopy imaging. CLEC19A expression in glioma cells and tissues was evaluated by qRT-PCR. Cell viability, proliferation, migration, and apoptosis were examined through MTT assay, CFSE assay, colony formation, wound healing assay, transwell test, and flow cytometry respectively after CLEC19A overexpression. The effect of CLEC19A overexpression on the PI3K/AKT/NF-κB signaling pathway was investigated using Western blot. An in vivo experiment substantiated the in vitro results using the glioblastoma rat models. Our in-silico analysis using TCGA data and measuring CLEC19A expression level by qRT-PCR determined significantly lower expression of CLEC19A in human glioma tissues compared to healthy brain tissues. By employment of ICC/IF, confocal microscopy imaging, and Western blot we could show that CLEC19A is plausibly a secreted protein. Results obtained from several in vitro readouts showed that CLEC19A overexpression in U87 and C6 glioma cell lines is associated with the inhibition of cell proliferation, viability, and migration. Further, qRT-PCR and Western blot analysis showed CLEC19A overexpression could reduce the expression levels of PI3K, VEGFα, MMP2, and NF-κB and increase PTEN, TIMP3, RECK, and PDCD4 expression levels in glioma cell lines. Furthermore, flow cytometry results revealed that CLEC19A overexpression was associated with significant cell cycle arrest and promotion of apoptosis in glioma cell lines. Interestingly, using a glioma rat model we could substantiate that CLEC19A overexpression suppresses glioma tumor growth. To our knowledge, this is the first report providing in-silico, molecular, cellular, and in vivo evidences on the role of CLEC19A as a putative tumor suppressor gene in GBM. These results enhance our understanding of the role of CLEC19A in glioma and warrant further exploration of CLEC19A as a potential therapeutic target for GBM.

Expression of brain-derived neurotrophic factor and formation of migrasome increases in the glioma cells induced by the adipokinetic hormone.

It has been previously shown that brain-derived neurotrophic factor is linked with various types of cancer. Brain-derived neurotrophic factor is found to be highly expressed in multiple human cancers and associated with tumor growth, invasion, and metastasis. Adipokinetic hormones are functionally related to the vertebrate glucagon, as they have similar functionalities that manage the nutrient-dependent secretion of these two hormones. Migrasomes are new organelles that contain numerous small vesicles, which aid in transmitting signals between the migrating cells. Therefore, the aim of this study was to investigate the effects of Anax imperator adipokinetic hormone on brain-derived neurotrophic factor expression and ultrastructure of cells in the C6 glioma cell line. The rat C6 glioma cells were treated with concentrations of 5 and 10 Anax imperator adipokinetic hormone for 24 h. The effects of the Anax imperator adipokinetic hormone on the migrasome formation and brain-derived neurotrophic factor expression were analyzed using immunocytochemistry and transmission electron microscope. The rat C6 glioma cells of the 5 and 10 μM Anax imperator adipokinetic hormone groups showed significantly high expressions of brain-derived neurotrophic factor and migrasomes numbers, compared with the control group. A positive correlation was found between the brain-derived neurotrophic factor expression level and the formation of migrasome, which indicates that the increased expression of brain-derived neurotrophic factor and the number of migrasomes may be involved to metastasis of the rat C6 glioma cell line induced by the Anax imperator adipokinetic hormone. Therefore, the expression of brain-derived neurotrophic factor and migrasome formation may be promising targets for preventing tumor proliferation, invasion, and metastasis in glioma.

Investigation of the Effect of Indatraline on Oxidative Damage Induced by Hydrogen Peroxide in C6 Glioma Cell Line

Oxidative stress is defined as an imbalance between the generation of reactive oxygen species (ROS) and their scavenging. Indatralin, which has serotonin reuptake inhibitory activity, has not yet been studied for its ability to prevent oxidative damage. Our research's objective was to find out how indatraline defends against oxidative damage. C6 cells were used in the study and four different cell groups were created. The control group received no therapy at all. For 24 hours, cells in the H2O2 group were exposed to 0.5 mM H2O2. The indatraline group received indatraline treatments for 24 hours at various doses (0.5, 1, 2.5, 5 and 10 μM). For one hour, indatraline was administered to the indatraline + H2O2 group at various concentrations (0.5, 1, 2.5, 5 and 10 μM) before the group was subjected to 0.5 mM H2O2 for 24 hours. Following the occurrence of oxidative damage, total antioxidant status (TAS) and total oxidant status (TOS) levels were determined. Cell viability was also evaluated using the XTT assay. As a result, after hydrogen peroxide-induced oxidative damage, indatraline at doses of 10, 5, and 2.5 μM showed a protective effect by significantly enhanced cell survival in C6 cells(p < 0.001). Additionally, indatraline boosted the lowered TAS level while decreasing the elevated TOS levels following hydrogen peroxide-induced oxidative damage (p<0.001).

Neurotrophic phenolic glycosides from the roots of Armoracia rusticana

Armoracia rusticana P. G. Gaertner. belongs to the Brassicaceae family and has aroused scientific interest for its anti-inflammatory and anticancer activities. In a continuing investigation to discover bioactive constituents from A. rusticana, we isolated 19 phenolic glycosides including three undescribed flavonol glycosides and one undescribed neolignan glycoside from MeOH extract of this plant. Their structures were elucidated based on NMR spectroscopic analysis (1H, 13C, 1H–1H COSY, HSQC, and HMBC), HRESIMS, and chemical methods. The determination of their absolute configuration was accomplished by ECD and LC-MS analysis. All the compounds were assessed for their potential neurotrophic activity through induction of nerve growth factor in C6 glioma cell lines and for their anti-neuroinflammatory activity based on the measurement of inhibition levels of nitric oxide production and pro-inflammatory cytokines (i.e., IL-1β, IL-6, and TNF-α) in lipopolysaccharide-activated microglia BV-2 cells.

In vivo C6 glioma models: an update and a guide toward a more effective preclinical evaluation of potential anti-glioblastoma drugs.

Glioblastoma multiform (GBM) is the most common primary brain tumor with a poor prognosis and few therapeutic choices. In vivo, tumor models are useful for enhancing knowledge of underlying GBM pathology and developing more effective therapies/agents at the preclinical level, as they recapitulate human brain tumors. The C6 glioma cell line has been one of the most widely used cell lines in neuro-oncology research as they produce tumors that share the most similarities with human GBM regarding genetic, invasion, and expansion profiles and characteristics. This review provides an overview of the distinctive features and the different animal models produced by the C6 cell line. We also highlight specific applications of various C6 invivo models according to the purpose of the study and offer some technical notes for more convenient/repeatable modeling. This work also includes novel findings discovered in our laboratory, which would further enhance the feasibility of the model in preclinical GBM investigations.

Surface-Modified Biobased Polymeric Nanoparticles for Dual Delivery of Doxorubicin and Gefitinib in Glioma Cell Lines.

Glioma is a malignant form of brain cancer that is challenging to treat due to the progressive growth of glial cells. To target overexpressed folate receptors in glioma brain tumors, we designed and investigated doxorubicin-gefitinib nanoparticles (Dox-Gefit NPs) and folate conjugated Dox-Gefit NPs (Dox-Gefit NPs-F). Dox-Gefit NPs and Dox-Gefit NPs-F were characterized by multiple techniques including Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), differential scanning calorimetry (DSC), proton nuclear magnetic resonance (1H NMR), and transmission electron microscopy (TEM). In vitro release profiles were measured at both physiological and tumor endosomal pH. The cytotoxicity of the Dox-Gefit NP formulations was measured against C6 and U87 glioma cell lines. A hemolysis assay was performed to investigate biocompatibility of the formulations, and distribution of the drugs in different organs was also estimated. The Dox-Gefit NPs and Dox-Gefit NPs-F were 109.45 ± 7.26 and 120.35 ± 3.65 nm in size and had surface charges of -18.0 ± 3.27 and -20.0 ± 8.23 mV, respectively. Dox-Gefit NPs and Dox-Gefit NPs-F significantly reduced the growth of U87 cells, with IC50 values of 9.9 and 3.2 μM. Similarly, growth of the C6 cell line was significantly reduced, with IC50 values of 8.43 and 3.31 μM after a 24 h incubation, in Dox-Gefit NPs and Dox-Gefit NPs-F, respectively. The percentage drug releases of Dox and Gefit from Dox-Gefit NPs at pH 7.4 were 60.87 ± 0.59 and 68.23 ± 0.1%, respectively. Similarly, at pH 5.4, Dox and Gefit releases from NPs were 70.87 ± 0.28 and 69.24 ± 0.12%, respectively. Biodistribution analysis revealed that more Dox and Gefit were present in the brain than in the other organs. The functionalized NPs inhibited the growth of glioma cells due to high drug concentrations in the brain. Folate conjugated NPs of Dox-Gefit could be a treatment option in glioma therapy.

Synthesis, physical properties and cytotoxic assessment of ester-terminated gemini imidazolium surfactants

Ester-functionalized gemini imidazolium amphiphiles appended with different spacers have been developed. The desired structures were confirmed using mass spectroscopy and NMR techniques. With the help of tensiometry, conductometry, and spectrofluorometry, the investigated amphiphiles have been characterized for a variety of physical properties including Gibbs free energy of adsorption (ΔG0ads), C20 (surfactant concentration required to reduce the surface tension of the solvent by 20 mN•m−1), maximum surface excess (Ʈmax), critical micelle concentration (cmc), Gibbs free energy of the micellization (ΔG0mic) and Amin (minimum surface area/molecule). Dynamic light scattering (DLS) measurements were taken to examine aggregate size distribution and zeta potential. Thermal decomposition temperature of investigated amphiphiles was determined from thermogravimetric analysis (TGA) analysis. By using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, the toxicity of examined amphiphiles towards the C6 glioma cell line was assessed. The physicochemical study of these surfactants revealed that varying spacer length considerably affects the properties.

Unveiling the Anti-cancer Potential of Bergenia ciliata (haw.) Sternb: A Mechanistic Study on UPR Modulation and ROS Generation

Bergenia ciliata (haw.) Sternb, a well-known medicinal plant, has traditionally been used to treat various ailments, such as diabetes, microbial infections, and kidney stones, owing to its anti-inflammatory and anti-tussive properties. Here we aimed to evaluate the anti-cancer potential of the plant extract and deduce the molecular pathways involved. To investigate this, we used the MTT assay to determine the IC50 values of the methanolic extract of B. ciliata (BcME) in MDA-MB-231 and C6-Glioma cell lines. After treating the cancer cell lines with BcME for 18 hours, we evaluated the UPR signalling markers, including pIRE1, Xbp1, ATF6, eIF2α, ATF4, and CHOP, using Immunoblotting. We also used RT-PCR to determine the mRNA levels of ATF4 and performed enzyme assays using Spectrophotometric techniques to measure ROS levels. Our results indicate that B. ciliata has potent anti-cancer properties due to its ability to modulate UPR and ROS pathways. Specifically, the plant extract effectively repressed the cytoprotective UPR by blocking IRE1-Xbp1 and ATF6 pathways while enhancing the PERK-ATF4-CHOP pathway, which is known to switch UPR towards apoptosis. Furthermore, our study revealed that B. ciliata significantly caused the accumulation of ROS in cancer cells and inhibited the antioxidant enzymes catalase and superoxide dismutase. These results conclusively suggest that BcME works synergistically on both UPR and ROS pathways to promote apoptosis and eliminate cancerous cells and thus serves as a potential primary source for bioactive molecules, selectively targeting cancer cells.

Swimming Training and Nanoformulation of Traditional Herbal Medicines Recovered Sensory-Motor Impairment by Modulating the Molecular Signaling Pathway in Rat Brain Tumor Models

Background: Glioblastoma (GBM) is one of the most aggressive tumors, health-threatening, and resistant to available treatment approaches for people worldwide. Objectives: This study aimed to evaluate effects of swimming training and a supplement comprising four nanoliposome herbal extracts in a rat model with mid-brain tumor. Methods: In this experimental study, 56-male-Wistar rats (230 ± 20 g) were divided into eight groups (n = 7): Normal group, model group, model + exe group, model + lipo, model + extract, model + lipo-extract, model + extract-exe, model + lipo-extract + exe. Mid-brain tumor model was induced by injection of the C6 glioma cell line (5 × 105 cell suspension) using stereotaxic techniques in the substantia nigra area of rats. Consumption of nanoformulation of herbals extract (100 mg/kg/day), crude extract (100 mg/kg/day), and swimming training (30 min, 3 days/week) were performed for six weeks. P53, Hras, IL-10, and Casp8 were studied by bioinformatics analysis. We assessed the relative expression of genes in the mid-brain tumor via the quantitative Real-Time PCR (qRT-PCR) method. Beam test and sciatic functional index assessed nerve function and motor coordination. Results: In silico analysis highlighted substantial genes with significant differential expression compared to healthy status as biomarkers in pathogenesis conditions. The expression level of the P53 and Hras increased with swimming training and nanoliposomes enriched complex complement. Besides, the expression of the IL-10 and Casp8 decreased with swimming training and nanoliposomes enriched complex complement. Conclusions: Our findings show that swimming training and nanoliposome-enriched combined supplements in rats can possibly be considered effective complementary medicine for motor impairment recovery induced by the brain tumor in the substantia nigra area.

The effect of hydro alcoholic extract of Nigella sativa seeds on inflammatory mediators in C6 glioma cell line

Nigella sativa (N. sativa) is a highly valued nutritional plant, which has long been used in traditional medicine to treat a variety of human diseases. The multifaceted pharmacological impacts of N. sativa, such as attenuating oxidative stress and inflammation, make it a suitable therapeutic candidate against cardiovascular, hepatic, and neurological disorders as well as cancer. Therefore, the current study aimed to evaluate the effect of the hydroalcoholic extract of N.sativa seeds on several pro-inflammatory cytokines in the C6 glioma cell line and to compare it with the effect of the extract on the normal fibroblast cell line. C6 and fibroblast cell lines were treated with the extract of N.sativa seeds, and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay was performed to determine the half-maximal inhibitory concentration (IC50) after 72h of treatment. Real-time polymerase chain reaction (RT-PCR) was carried out to assess the expression levels of interleukin (IL)-6, IL-10, tumor necrosis factor-alpha (TNF-α), and transforming growth factor- β1 (TGF-β1) at the mRNA level in both cell lines after 72h of treatment with non-toxic and IC50 concentrations obtained from C6 cell line. The IC50 values for the hydroalcoholic extract of N.sativa seeds were 260±20μg/mL in the C6 cell line and 398±27μg/mL in fibroblast cells. The real-time PCR results indicated that the treatment of C6 and fibroblast cells with the extract at the IC50 value of N.sativa in C6 for 72h could increase the mRNA expression levels of IL-10 and reduce the mRNA expression levels of IL-6, TNF-α, and TGF-β1 in C6 and fibroblast cells. The N.sativa extract showed a higher anti-inflammatory effect on C6 cells in comparison with fibroblast cells. Regarding the anti-inflammatory effect of Nigella sativa in C6 cell line, it may be considered a promising candidate to fortify antitumor actions in combination with other therapeutic options in the treatment of patients with GBM.

Exposure of aluminium to C6 glioma cells modulates molecular and functional neurotoxic markers.

The risk of aluminium exposure to humans is very high as it may get into the human body through excessive dietary contaminants, inhalation of fine particulate matter, or through parenteral routes as a vaccine adjuvant and so forth. The increased level of aluminium in brain tissue has been shown to be associated with several neurodegenerative and neurotoxic adverse effects, including AD. However, the exact mechanism of aluminium-induced neurotoxicity is still unclear. Therefore, our study aimed to investigate the mechanism of neurotoxic and neurodegenerative effects through in vitro exposure of aluminium in rat glioma C6 cell line. The findings of our study have indicated that aluminium chloride exposure may lead to alteration in acetylcholine levels, increased oxidative imbalanceand induction of molecular structural and functional markers of neuronal inflammation. This study also demonstrated that aluminium exposure may lead to the induction of caspase-3 along with apoptotic cell death and a significant increase in amyloid-beta and hyperphosphorylated tau levels in C6 cells. Thus, this study may provide a mechanistic understanding of the regulation of neuroinflammatory and neurodegenerative biomarkers due to aluminium exposure.

Anti-Proliferative Potential of Quercetin Loaded Polymeric Mixed Micelles on Rat C6 and Human U87MG Glioma Cells.

Quercetin (Qu) is a natural flavonoid present in many commonly consumed food items and is also identified as a potential anticancer agent. The present study evaluates the Qu-loaded polymeric mixed micelles (Qu-PMMs) against C6 and U87MG glioma cell lines. The Box–Behnken Design (BBD) was employed to study the influence of independent variables such as Soluplus, Vitamin-E polyethyleneglycol-1000 succinate (E-TPGS), and poloxamer 407 concentrations on dependent variables including particle size (PS), polydispersity index (PDI), and percentage entrapment efficiency (%EE) of the prepared Qu-PMMs. The Qu-PMMs were further characterized by Fourier Transform Infrared Spectroscopy (FTIR), X-ray Diffraction (XRD), Scanning Electron Microscope (SEM), and were assessed for in vitro drug release, effect on cell viability, migration, cellular uptake, and apoptosis assays. The PS, PDI, and % EE of the optimized PMMs were 107.16 ± 1.06 nm, 0.236 ± 0.053, and 77.46 ± 1.94%, respectively. The FTIR and XRD revealed that the Qu was completely entrapped inside the PMMs. The SEM analysis confirmed the spherical shape of micelles. The in vitro cell viability study showed that the Qu-PMMs had 1.7 times higher cytotoxicity against C6 and U87MG cells than Qu pure drug (Qu-PD). Furthermore, Qu-PMMs demonstrated superior cellular uptake, inhibited migration, and induced apoptosis when tested against C6 and U87MG cells than pure Qu. Thus, the polymeric mixed micelle (PMMs) enhanced the therapeutic effect of Qu and can be considered an effective therapeutic strategy to treat Glioma.