Water Extract from the Leaves of Withania somnifera Protect RA Differentiated C6 and IMR-32 Cells against Glutamate-Induced Excitotoxicity

Department of Biotechnology, Guru Nanak Dev University, Amritsar, India.
PLoS ONE (Impact Factor: 3.23). 05/2012; 7(5):e37080. DOI: 10.1371/journal.pone.0037080
Source: PubMed


Glutamate neurotoxicity has been implicated in stroke, head trauma, multiple sclerosis and neurodegenerative disorders. Search for herbal remedies that may possibly act as therapeutic agents is an active area of research to combat these diseases. The present study was designed to investigate the neuroprotective role of Withania somnifera (Ashwagandha), also known as Indian ginseng, against glutamate induced toxicity in the retinoic acid differentiated rat glioma (C6) and human neuroblastoma (IMR-32) cells. The neuroprotective activity of the Ashwagandha leaves derived water extract (ASH-WEX) was evaluated. Cell viability and the expression of glial and neuronal cell differentiation markers was examined in glutamate challenged differentiated cells with and without the presence of ASH-WEX. We demonstrate that RA-differentiated C6 and IMR-32 cells, when exposed to glutamate, undergo loss of neural network and cell death that was accompanied by increase in the stress protein HSP70. ASH-WEX pre-treatment inhibited glutamate-induced cell death and was able to revert glutamate-induced changes in HSP70 to a large extent. Furthermore, the analysis on the neuronal plasticity marker NCAM (Neural cell adhesion molecule) and its polysialylated form, PSA-NCAM revealed that ASH-WEX has therapeutic potential for prevention of neurodegeneration associated with glutamate-induced excitotoxicty.

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Available from: Hardeep Kataria, Oct 01, 2015
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    • "In another study, it was demonstrated that W. somnifera root extract and withanolide-A were capable of restoring spatial memory deficit by inhibiting oxidative stress induced alteration in glutamergic neurotransmission, where W. somnifera reduces the expression of N-metyl-D-aspartate (NMDA) receptor, which is responsible for spatial memory loss in epileptic rats (Soman et al., 2012). Leaf extracts of W. somnifera were also showing its protective action against glutamate induced toxicity in human neuroblastoma (IMR-32) cells, by inhibiting over expression of stress protein 70 kilodalton heat shock proteins (Kataria et al., 2012). In another study it was found that W. somnifera root extract and withanolide-A regulate the expression and function of α-Amino-3-hydroxy-5-methyl-4- isoxazolepropionic acid receptor and glutamate levels in brain dopaminergic nervous system and results are attributed to improvement in motor learning in pilocarpine-induced temporal lobe epilepsy model (Soman et al., 2013). "
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    ABSTRACT: Withania somnifera (Ashwagandha) is an important Rasayana herb and widely considered as Indian ginseng in Ayurveda. In traditional system of Indian medicine, it is used as tonic to rejuvenate the body and increase longevity. In Ayurvedic preparations, various parts of the plant have been used to treat variety of ailments that affect the human health. However, dried roots of the plant are widely used for the treatment of nervous and sexual disorders. The major active chemical constituents of this plant are withanolides, which is responsible for its wide range of biological activities. Since the beginning of the 20 th century, a significant amount of research has been done and efforts are ongoing to further explore other bioactive constituents, and many pharmacological studies have been carried out to describe their disease preventing mechanisms. In this chapter, we have reviewed the chemistry and pharmacological basis of W. somnifera in various human ailments.
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    • "Consistent with these reports, expression of NCAM was found to decrease in lead nitrate exposed C6 cells, whereas Ashwagandha treatment was observed to augment the lead mediated downregulation of NCAM partially as shown by immunostaining and RT-PCR. A recent study on Ashwagandha treatment in C6 glioma cells has shown significant increase in expression of NCAM [39] and also increase in NCAM expression after Ashwagandha treatment has been associated with protection against glutamate induced damage in RA differentiated neuronal cultures [34]. "
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    ABSTRACT: Withania somnifera (Ashwagandha), also known as Indian Ginseng, is a well-known Indian medicinal plant due to its antioxidative, antistress, antigenotoxic, and immunomodulatory properties. The present study was designed to assess and establish the cytoprotective potential of Ashwagandha leaf aqueous extract against lead induced toxicity. Pretreatment of C6 cells with 0.1% Ashwagandha extract showed cytoprotection against 25 μM to 400 μM concentration of lead nitrate. Further pretreatment with Ashwagandha extract to lead nitrate exposed cells (200 μM) resulted in normalization of glial fibrillary acidic protein (GFAP) expression as well as heat shock protein (HSP70), mortalin, and neural cell adhesion molecule (NCAM) expression. Further, the cytoprotective efficacy of Ashwagandha extract was studied in vivo. Administration of Ashwagandha extract provided significant protection to lead induced altered antioxidant defense that may significantly compromise normal cellular function. Ashwagandha also provided a significant protection to lipid peroxidation (LPx) levels, catalase, and superoxide dismutase (SOD) but not reduced glutathione (GSH) contents in brain tissue as well as peripheral organs, liver and kidney, suggesting its ability to act as a free radical scavenger protecting cells against toxic insult. These results, thus, suggest that Ashwagandha water extract may have the potential therapeutic implication against lead poisoning.
    BioMed Research International 04/2014; 2014. DOI:10.1155/2014/182029 · 3.17 Impact Factor
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    • "Although the use of various compounds like retinoids, taxol, paclitaxel, and PKC inhibitors have been shown to have therapeutic potential but the very nature of glioma exhibiting resistance against chemotherapy and radiotherapy demands new therapeutic drugs [79]. Differentiation inducing effect of TCE was found to be comparable to the effect of ATRA (All Trans-Retinoic Acid) that we have used to induce differentiation in C6 cells in our previous studies [80]. Current data suggests that TCE may have the potential to induce differentiation in C6 glioma cells by targeting different pathways related to cell proliferation, differentiation, senescence and ultimately apoptosis. "
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    ABSTRACT: Glioblastomas are the most aggressive primary brain tumors and their heterogeneity and complexity often renders them non responsive to various conventional treatments. Search for herbal products having potential anti-cancer activity is an active area of research in the Indian traditional system of medicine i.e., Ayurveda. Tinospora cordifolia, also named as 'heavenly elixir' is used in various ayurvedic decoctions as panacea to treat several body ailments. The current study investigated the anti-brain cancer potential of 50% ethanolic extract of Tinospora cordifolia (TCE) using C6 glioma cells. TCE significantly reduced cell proliferation in dose-dependent manner and induced differentiation in C6 glioma cells, resulting in astrocyte-like morphology as indicated by phase contrast images, GFAP expression and process outgrowth data of TCE treated cells which exhibited higher number and longer processes than untreated cells. Reduced proliferation of cells was accompanied by enhanced expression of senescence marker, mortalin and its translocation from perinuclear to pancytoplasmic spaces. Further, TCE showed anti-migratory and anti-invasive potential as depicted by wound scratch assay and reduced expression of plasticity markers NCAM and PSA-NCAM along with MMP-2 and 9. On analysis of the cell cycle and apoptotic markers, TCE treatment was seen to arrest the C6 cells in G0/G1 and G2/M phase, suppressing expression of G1/S phase specific protein cyclin D1 and anti-apoptotic protein Bcl-xL, thus supporting its anti-proliferative and apoptosis inducing potential. Present study provides the first evidence for the presence of anti-proliferative, differentiation-inducing and anti-migratory/anti-metastatic potential of TCE in glioma cells and possible signaling pathways involved in its mode of action. Our primary data suggests that TCE and its active components may prove to be promising phytotherapeutic interventions in gliobalstoma multiformae.
    PLoS ONE 10/2013; 8(10):e78764. DOI:10.1371/journal.pone.0078764 · 3.23 Impact Factor
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