Investigation of Antioxidant Efficacy of Glycyrrhiza glabra L. Extract in Glutamate Toxicity-Induced Primary Neuron Culture

Yıl 2022, Cilt: 3 Sayı: 1, 18 - 24, 30.06.2022

Cemil Bayram Ahmet Hacimuftuoglu

Öz

Background: Glutamate toxicity and reactive oxygen accumulation are life-threatening factors that play a role in the pathogenesis of neurodegenerative diseases. Today, many studies are carried out to elucidate these pathological pathways and to reveal possible treatment possibilities. Herbal medicines have been used continuously by people since ancient times due to their low side effects and easy accessibility. Glycyrrhiza glabra L. extract is also one of the plants often used for medicinal purposes. Since the antioxidant activity of Glycyrrhiza glabra L. has been reported in the studies, its protective activity at different doses was investigated in primary neuron culture with glutamate toxicity.

Methodology/Main findings: In the experiment, which consisted of control, Glutamate Toxicity, GG 100 µg/ml, GG 200 µg/ml and GG 400 µg/ml experimental groups, primary neuron culture was performed and some tests were applied in order to test cell viability and antioxidant activity in this culture.These tests are superoxide dismutase, total antioxidant level, total oxidant level, malondialhedit level, lactate dehydrogenase activity and MTT (3-(4,5-dimethyltriazol-2-yl)-2,5-diphenyltetrazolium bromide) tests. Our GG 200 µg/ml treatment group had the most significant results in all tests in statistical studies. In all tests, the glutamate toxicity group was compared with the Control group, and the treatment groups were statistically compared with the glutamate toxicity group.

Conclusion: Significant results were obtained in the tests applied to determine the oxidative stress and cell viability caused by glutamate toxicity. In the study, it was shown that Glycyrrhiza glabra L. significantly reduced glutamate-induced cell death, LDH release, lipid peroxidation and oxidation in primary neuronal cells.

Anahtar Kelimeler

Glutamate toxicity, Glycyrrhiza glabra L., Liquorice, Oxidative stress, Primer Neuron culture

Kaynakça

• M. Ahvazi, F. Khalighi-Sigaroodi, M. M. Charkhchiyan, F. Mojab, V.-A. Mozaffarian, and H. Zakeri, "Introduction of medicinal plants species with the most traditional usage in Alamut region," Iranian journal of pharmaceutical research: IJPR, vol. 11, no. 1, p. 185, 2012.
• S. Nazari, M. Rameshrad, and H. Hosseinzadeh, "Toxicological effects of Glycyrrhiza glabra (licorice): a review," Phytotherapy research, vol. 31, no. 11, pp. 1635-1650, 2017.
• P. Skehan et al., "New colorimetric cytotoxicity assay for anticancer-drug screening," JNCI: Journal of the National Cancer Institute, vol. 82, no. 13, pp. 1107-1112, 1990.
• G. Pastorino, L. Cornara, S. Soares, F. Rodrigues, and M. B. P. Oliveira, "Liquorice (Glycyrrhiza glabra): A phytochemical and pharmacological review," Phytotherapy research, vol. 32, no. 12, pp. 2323-2339, 2018.
• B. Soylemez and S. Sahin-Bolukbasi, "Evaluation of the Neurotoxicity of Strontium and Glycyrrhiza Glabra: First Report," (in eng), Turk Neurosurg, vol. 31, no. 4, pp. 505-509, 2021, doi: 10.5137/1019-5149.Jtn.30290-20.2.
• A. Hacımüftüoğlu, U. Okkay, M. Sağsöz, and M. Taşpınar, "DENEYSEL ALZHEİMER MODELİNDE HİPOKAMPÜSTE GLUTAMAT GERİ ALINIM PARAMETRELERİNİN DEĞERLENDİRİLMESİ," 2021.
• A. Lau and M. Tymianski, "Glutamate receptors, neurotoxicity and neurodegeneration," Pflügers Archiv-European Journal of Physiology, vol. 460, no. 2, pp. 525-542, 2010.
• S. M. Rothman and J. W. Olney, "Glutamate and the pathophysiology of hypoxic–ischemic brain damage," Annals of Neurology: Official Journal of the American Neurological Association and the Child Neurology Society, vol. 19, no. 2, pp. 105-111, 1986.
• H. Chi, H. Y. Chang, and T. K. Sang, "Neuronal Cell Death Mechanisms in Major Neurodegenerative Diseases," (in eng), Int J Mol Sci, vol. 19, no. 10, Oct 9 2018, doi: 10.3390/ijms19103082.
• A. Singh, R. Kukreti, L. Saso, and S. Kukreti, "Oxidative Stress: A Key Modulator in Neurodegenerative Diseases," (in eng), Molecules, vol. 24, no. 8, Apr 22 2019, doi: 10.3390/molecules24081583.
• A. A. Kritis, E. G. Stamoula, K. A. Paniskaki, and T. D. Vavilis, "Researching glutamate - induced cytotoxicity in different cell lines: a comparative/collective analysis/study," (in eng), Front Cell Neurosci, vol. 9, p. 91, 2015, doi:10.3389/fncel.2015.00091.
• A. Federico, E. Cardaioli, P. Da Pozzo, P. Formichi, G. N. Gallus, and E. Radi, "Mitochondria, oxidative stress and neurodegeneration," (in eng), J Neurol Sci, vol. 322, no. 1-2, pp. 254-62, Nov 15 2012, doi: 10.1016/j.jns.2012.05.030.
• T. M. Dawson, V. L. Dawson, and S. H. Snyder, "A novel neuronal messenger molecule in brain: the free radical, nitric oxide," (in eng), Ann Neurol, vol. 32, no. 3, pp. 297-311, Sep 1992, doi: 10.1002/ana.410320302.
• A. Dumuis, M. Sebben, L. Haynes, J. P. Pin, and J. Bockaert, "NMDA receptors activate the arachidonic acid cascade system in striatal neurons," (in eng), Nature, vol. 336, no. 6194, pp. 68-70, Nov 3 1988, doi: 10.1038/336068a0.
• J. M. McCord, "Oxygen-derived free radicals in postischemic tissue injury," (in eng), N Engl J Med, vol. 312, no. 3, pp. 159-63, Jan 17 1985, doi: 10.1056/nejm198501173120305.
• S. Bannai and E. Kitamura, "Transport interaction of L-cystine and L-glutamate in human diploid fibroblasts in culture," (in eng), J Biol Chem, vol. 255, no. 6, pp. 2372-6, Mar 25 1980.
• I. Ahiskali et al., "Effect of taxifolin on cisplatin-associated oxidative optic nerve damage in rats," Cutaneous and Ocular Toxicology, vol. 40, no. 1, pp. 1-6, 2021.
• U. Okkay and I. F. Okkay, "Beneficial effects of linagliptin in cell culture model of Parkinson’s disease," The European Research Journal, vol. 8, no. 2, pp. 242-246, 2022.
• U. Okkay and I. F. Okkay, "In vitro neuroprotective effects of allicin on Alzheimer’s disease model of neuroblastoma cell line," Journal of Surgery and Medicine, vol. 6, no. 2, pp. 209-212, 2022.
• U. Okkay and I. F. Okkay, "Neuroprotective effect of nicorandil in 6-OHDA induced in vitro model of parkinson's disease," Medicine, vol. 11, no. 1, pp. 224-7, 2022.
• B. Sadeghzadeh Oskouei, S. Zargari, P. Shahabi, M. Ghaffari Novin, and M. Pashaiasl, "Design and Microfabrication of An On-Chip Oocyte Maturation System for Reduction of Apoptosis," (in eng), Cell J, vol. 23, no. 1, pp. 32-39, 2021, doi: 10.22074/cellj.2021.7056.
• B. Rasool Hassan, "Medicinal plants (importance and uses)," Pharmaceut Anal Acta, vol. 3, no. 10, pp. 2153-2435, 2012.
• V. P. Kamboj, "Herbal medicine," Current science, vol. 78, no. 1, pp. 35-39, 2000.
• D. Sharma, P. Namdeo, and P. Singh, "Phytochemistry & Pharmacological Studies of Glycyrrhiza glabra: A Medicinal Plant Review," Int. J. Pharm. Sci. Rev. Res., vol. 67, pp. 187-194, 2021.
• G. E. Batiha, A. M. Beshbishy, D. S. Tayebwa, O. S. Adeyemi, N. Yokoyama, and I. Igarashi, "Evaluation of the inhibitory effect of ivermectin on the growth of Babesia and Theileria parasites in vitro and in vivo," (in eng), Trop Med Health, vol. 47, p. 42, 2019, doi: 10.1186/s41182-019-0171-8.
• G. E.-S. BATIHA, A. M. BESHBISHY, D. S. TAYEBWA, H. M. SHAHEEN, N. YOKOYAMA, and I. IGARASHI, "Inhibitory effects of Uncaria tomentosa bark, Myrtus communis roots, Origanum vulgare leaves and Cuminum cyminum seeds extracts against the growth of Babesia and Theileria in vitro," Jpn. J. Vet. Parasitol. Vol, vol. 17, no. 1, 2018.
• A. M. Beshbishy, G. E.-S. Batiha, O. S. Adeyemi, N. Yokoyama, and I. Igarashi, "Inhibitory effects of methanolic Olea europaea and acetonic Acacia laeta on growth of Babesia and Theileria," Asian Pacific Journal of Tropical Medicine, vol. 12, no. 9, p. 425, 2019.
• D. M. Biondi, C. Rocco, and G. Ruberto, "New Dihydrostilbene Derivatives from the Leaves of Glycyrrhiza g labra and Evaluation of Their Antioxidant Activity," Journal of natural products, vol. 66, no. 4, pp. 477-480, 2003.
• R. Isbrucker and G. Burdock, "Risk and safety assessment on the consumption of Licorice root (Glycyrrhiza sp.), its extract and powder as a food ingredient, with emphasis on the pharmacology and toxicology of glycyrrhizin," Regulatory Toxicology and Pharmacology, vol. 46, no. 3, pp. 167-192, 2006.
• J. Cinatl, B. Morgenstern, G. Bauer, P. Chandra, H. Rabenau, and H. Doerr, "Glycyrrhizin, an active component of liquorice roots, and replication of SARS-associated coronavirus," The Lancet, vol. 361, no. 9374, pp. 2045-2046, 2003.
• M. Ito et al., "Inhibitory effect of glycyrrhizin on the in vitro infectivity and cytopathic activity of the human immunodeficiency virus [HIV (HTLV-III/LAV)]," (in eng), Antiviral Res, vol. 7, no. 3, pp. 127-37, Mar 1987, doi: 10.1016/0166-3542(87)90001-5.
• S. Rahman and S. Sultana, "Chemopreventive activity of glycyrrhizin on lead acetate mediated hepatic oxidative stress and its hyperproliferative activity in Wistar rats," (in eng), Chem Biol Interact, vol. 160, no. 1, pp. 61-9, Mar 10 2006, doi: 10.1016/j.cbi.2005.12.003.
• H. Sato et al., "Therapeutic basis of glycyrrhizin on chronic hepatitis B," (in eng), Antiviral Res, vol. 30, no. 2-3, pp. 171-7, May 1996, doi: 10.1016/0166-3542(96)00942-4.
• T. Yoshida, Y. Tsuda, D. Takeuchi, M. Kobayashi, R. B. Pollard, and F. Suzuki, "Glycyrrhizin inhibits neutrophil-associated generation of alternatively activated macrophages," Cytokine, vol. 33, no. 6, pp. 317-322, 2006.
• R. Iida, Y. Otsuka, K. Matsumoto, S. Kuriyama, and T. Hosoya, "Pseudoaldosteronism due to the concurrent use of two herbal medicines containing glycyrrhizin: interaction of glycyrrhizin with angiotensin-converting enzyme inhibitor," (in eng), Clin Exp Nephrol, vol. 10, no. 2, pp. 131-5, Jun 2006, doi: 10.1007/s10157-006-0415-x.
• B. Jayaprakasam, S. Doddaga, R. Wang, D. Holmes, J. Goldfarb, and X. M. Li, "Licorice flavonoids inhibit eotaxin-1 secretion by human fetal lung fibroblasts in vitro," (in eng), J Agric Food Chem, vol. 57, no. 3, pp. 820-5, Feb 11 2009, doi: 10.1021/jf802601j.
• E. Akbay, H. Arbağ, Y. Uyar, and K. Oztürk, "Oxidative stress and antioxidant factors in pathophysiology of allergic rhinitis," Kulak Burun Bogaz Ihtisas Dergisi: KBB= Journal of Ear, Nose, and Throat, vol. 17, no. 4, pp. 189-196, 2007.
• K. J. Kim, J. S. Choi, K. W. Kim, and J. W. Jeong, "The anti-angiogenic activities of glycyrrhizic acid in tumor progression," (in eng), Phytother Res, vol. 27, no. 6, pp. 841-6, Jun 2013, doi: 10.1002/ptr.4800.
• J. M. Cherng, H. J. Lin, M. S. Hung, Y. R. Lin, M. H. Chan, and J. C. Lin, "Inhibition of nuclear factor kappaB is associated with neuroprotective effects of glycyrrhizic acid on glutamate-induced excitotoxicity in primary neurons," (in eng), Eur J Pharmacol, vol. 547, no. 1-3, pp. 10-21, Oct 10 2006, doi: 10.1016/j.ejphar.2006.06.080.
• T. C. Kao, M. H. Shyu, and G. C. Yen, "Neuroprotective effects of glycyrrhizic acid and 18beta-glycyrrhetinic acid in PC12 cells via modulation of the PI3K/Akt pathway," (in eng), J Agric Food Chem, vol. 57, no. 2, pp. 754-61, Jan 28 2009, doi: 10.1021/jf802864k.
• L. Teng et al., "Liquiritin modulates ERK‑and AKT/GSK‑3β‑dependent pathways to protect against glutamate‑induced cell damage in differentiated PC12 cells," Molecular Medicine Reports, vol. 10, no. 2, pp. 818-824, 2014.
• K. Naveen and A. Bhattacharjee, "MEDICINAL HERBS AS NEUROPROTECTIVE AGENTS," 2021.
• K. K. Chakravarthi and R. Avadhani, "Beneficial effect of aqueous root extract of Glycyrrhiza glabra on learning and memory using different behavioral models: An experimental study," Journal of natural science, biology, and medicine, vol. 4, no. 2, p. 420, 2013.
• P. Hasanein, "Glabridin as a major active isoflavan from Glycyrrhiza glabra (licorice) reverses learning and memory deficits in diabetic rats," Acta Physiologica Hungarica, vol. 98, no. 2, pp. 221-230, 2011.
• S. M. Greenwood and C. N. Connolly, "Dendritic and mitochondrial changes during glutamate excitotoxicity," Neuropharmacology, vol. 53, no. 8, pp. 891-898, 2007.
• D. Schubert, "Signaling by reactive oxygen species in the nervous system," Cellular and Molecular Life Sciences CMLS, vol. 57, no. 8, pp. 1287-1305, 2000.
• M. Irani, M. Sarmadi, and F. Bernard, "Leaves antimicrobial activity of Glycyrrhiza glabra L," Iranian journal of pharmaceutical research: IJPR, vol. 9, no. 4, p. 425, 2010.
• S. Rodino, A. Butu, M. Butu, and P. Cornea, "Comparative studies on antibacterial activity of licorice, elderberry and dandelion," Digest Journal of Nanomaterials and Biostructures, vol. 10, no. 3, pp. 947-955, 2015.
• F. Sedighinia and A. S. Afshar, "Antibacterial activity of Glycyrrhiza glabra against oral pathogens: an in vitro study," Avicenna journal of phytomedicine, vol. 2, no. 3, p. 118, 2012.
• H. Prentice, J. P. Modi, and J.-Y. Wu, "Mechanisms of Neuronal Protection against Excitotoxicity, Endoplasmic Reticulum Stress, and Mitochondrial Dysfunction in Stroke and Neurodegenerative Diseases," Oxidative Medicine and Cellular Longevity, vol. 2015, p. 964518, 2015/10/20 2015, doi: 10.1155/2015/964518.
• A. Martínez et al., "Mutagenicity and cytotoxicity of fruits and vegetables evaluated by the Ames test and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay/Mutagenicidad y citotoxicidad de frutas y vegetales evaluadas por el test de Ames y el ensayo del bromuro de 3-(4, 5-dimetiltiazol-2-il)-2, 5-difeniltetrazolio (MTT)," Food science and technology international, vol. 5, no. 5, pp. 431-437, 1999.
• S. Khazraei-Moradian, M. Ganjalikhani-Hakemi, A. Andalib, R. Yazdani, J. Arasteh, and G. A. Kardar, "The effect of licorice protein fractions on proliferation and apoptosis of gastrointestinal cancer cell lines," Nutrition and cancer, vol. 69, no. 2, pp. 330-339, 2017.
• S. K. Lee, K. K. Park, J. H. Y. Park, S. S. Lim, and W. Y. Chung, "The inhibitory effect of roasted licorice extract on human metastatic breast cancer cell‐induced bone destruction," Phytotherapy Research, vol. 27, no. 12, pp. 1776-1783, 2013.
• S. M. Nourazarian, A. Nourazarian, M. Majidinia, and E. Roshaniasl, "Effect of root extracts of medicinal herb Glycyrrhiza glabra on HSP90 gene expression and apoptosis in the HT-29 colon cancer cell line," Asian Pacific journal of cancer prevention, vol. 16, no. 18, pp. 8563-8566, 2016.
• W. Song et al., "Uralsaponins M–Y, antiviral triterpenoid saponins from the roots of Glycyrrhiza uralensis," Journal of natural products, vol. 77, no. 7, pp. 1632-1643, 2014.
• V. M. Di Mambro and M. J. Fonseca, "Assays of physical stability and antioxidant activity of a topical formulation added with different plant extracts," Journal of Pharmaceutical and Biomedical Analysis, vol. 37, no. 2, pp. 287-295, 2005.
• P. Mutaillifu et al., "Structural characterization and antioxidant activities of a water soluble polysaccharide isolated from Glycyrrhiza glabra," International journal of biological macromolecules, vol. 144, pp. 751-759, 2020.
• M. Parle, D. Dhingra, and S. K. Kulkarni, "Memory-strengthening activity of Glycyrrhiza glabra in exteroceptive and interoceptive behavioral models," (in eng), J Med Food, vol. 7, no. 4, pp. 462-6, Winter 2004, doi: 10.1089/jmf.2004.7.462.
• L. Racková et al., "Mechanism of anti-inflammatory action of liquorice extract and glycyrrhizin," (in eng), Nat Prod Res, vol. 21, no. 14, pp. 1234-41, Dec 2007, doi: 10.1080/14786410701371280.
• S. Varsha, R. Agrawal, and P. Sonam, "Phytochemical screening and determination of anti-bacterial and anti-oxidant potential of Glycyrrhiza glabra root extracts," Journal of environmental Research and Development, vol. 7, no. 4A, p. 1552, 2013.
• V. Singh, A. Pal, and M. P. Darokar, "A polyphenolic flavonoid glabridin: Oxidative stress response in multidrug-resistant Staphylococcus aureus," (in eng), Free Radic Biol Med, vol. 87, pp. 48-57, Oct 2015, doi: 10.1016/j.freeradbiomed.2015.06.016.