TOXICOLOGICAL AND BEHAVIOR ANALYSES OF A FOLIOSE LICHEN PARMOTREMA TINCTORUM (DELISE EX NYL.) HALE EXTRACT USING ZEBRAFISH (DANIO RERIO) MODEL
DOI:
https://doi.org/10.22159/ajpcr.2021.v14i10.42134Keywords:
Lichen, Parmotrema tinctorum, Zebrafish, Toxicity, Nutraceutical effect, Histopathology, Inflammatory markersAbstract
Objective: The objective of the study was to evaluate the toxicity and behavior analysis with the extract of foliose lichen Parmotrema tinctorum against the adult zebrafish.
Methods: The zebrafishes were exposed to different concentrations of P. tinctorum extract such as 50, 100, and 200 mg/ml for 7 days of exposure, in which the mortality and behavioral responses of zebrafishes were recorded. The standard histopathological examination was conducted with the lichen extracts of P. tinctorum.
Results: The results revealed that P. tinctorum extract did not show any prominent behavior abnormalities in zebrafishes even at a high concentration of 200 mg/ml. The extract was found to have dose-dependent toxic to zebrafish and the number of neutrophil cells in the muscle bundles reduced at a high concentration. The results of the inflammatory marker gene expression using polymerase chain reaction results suggested that the dose-dependent suppression of tumor necrosis factor gene by the P. tinctorum lichen extract.
Conclusion: Overall, concluded that the extract might contain anti-inflammatory induction properties and further tests are required to prove apoptosis and anticancer activity using other in vivo or in vitro techniques.
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References
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[35] Matthias KA, Roche AM, Standish AJ, Shchepetov M, Weiser JN. Neutrophil-Toxin Interactions Promote Antigen Delivery and Mucosal Clearance of Streptococcus pneumoniae. J Immunol 2008;180:6246–54. https://doi.org/10.4049/jimmunol.180.9.6246.
[36] Woodfin A, Voisin M-B, Beyrau M, Colom B, Caille D, Diapouli F-M, et al. The junctional adhesion molecule JAM-C regulates polarized transendothelial migration of neutrophils in vivo. Nat Immunol 2011;12:761–769.
[37] Johnston IA, Bower NI, Macqueen DJ. Growth and the regulation of myotomal muscle mass in teleost fish. J Exp Biol 2011;214:1617–28. https://doi.org/10.1242/jeb.038620.
[38] Hong J-M, Kim JE, Min SK, Kim KH, Han SJ, Yim JH, et al. Anti-Inflammatory Effects of Antarctic Lichen Umbilicaria antarctica Methanol Extract in Lipopolysaccharide-Stimulated RAW 264.7 Macrophage Cells and Zebrafish Model. BioMed Res Int 2021;2021.
[2] Rodriguez CM, Bennett JP, Johnson CJ. Lichens: unexpected anti-prion agents? Prion 2012;6:11–16.
[3] Shukla V, Joshi GP, Rawat MSM. Lichens as a potential natural source of bioactive compounds: a review. Phytochem Rev 2010;9:303–314.
[4] Zambare VP, Christopher LP. Biopharmaceutical potential of lichens. Pharm Biol 2012;50:778–798.
[5] Crawford SD. Lichens Used in Traditional Medicine. In: Rankovi? B, editor. Lichen Second. Metab. Bioact. Prop. Pharm. Potential, Cham: Springer International Publishing; 2019, p. 31–97. https://doi.org/10.1007/978-3-030-16814-8_2.
[6] Ismail HF, Hashim Z, Soon WT, Rahman NSA, Zainudin AN, Majid FAA. Comparative study of herbal plants on the phenolic and flavonoid content, antioxidant activities and toxicity on cells and zebrafish embryo. J Tradit Complement Med 2017;7:452–65. https://doi.org/10.1016/j.jtcme.2016.12.006.
[7] Bencan Z, Sledge D, Levin ED. Buspirone, chlordiazepoxide and diazepam effects in a zebrafish model of anxiety. Pharmacol Biochem Behav 2009;94:75–80. https://doi.org/10.1016/j.pbb.2009.07.009.
[8] Benneh CK, Biney RP, Mante PK, Tandoh A, Adongo DW, Woode E. Maerua angolensis stem bark extract reverses anxiety and related behaviours in zebrafish-Involvement of GABAergic and 5-HT systems. J Ethnopharmacol 2017;207:129–45. https://doi.org/10.1016/j.jep.2017.06.012.
[9] Poornima S, Nagarjun N, Ponmurugan P, Gnanamangai BM, Narasimman S. Toxicity and anti-inflammatory study of Parmotrema austrosinense extract against oxozalone induced intestinal inflammation in zebrafish (Danio rerio) model. Biocatal Agric Biotechnol 2019;21:101278. https://doi.org/10.1016/j.bcab.2019.101278.
[10] Sztanke M, Rzymowska J, Sztanke K. Anticancer active trifluoromethylated fused triazinones are safe for early-life stages of zebrafish (Danio rerio) and reveal a proapoptotic action. J Enzyme Inhib Med Chem 2021;36:336–44. https://doi.org/10.1080/14756366.2020.1865944.
[11] Fernandes H, Peres H, Carvalho AP. Dietary protein requirement during juvenile growth of zebrafish (Danio rerio). Zebrafish 2016;13:548–555.
[12] Upreti DK, Divakar PK, Nayaka S. Notes on species of the lichen genus Phyllopsora in India. Bibl Lichenol 2002;86:185–191.
[13] Honda NK, Pavan FR, Coelho RG, de Andrade Leite SR, Micheletti AC, Lopes TIB, et al. Antimycobacterial activity of lichen substances. Phytomedicine 2010;17:328–32. https://doi.org/10.1016/j.phymed.2009.07.018.
[14] Tiwari P, Rai H, Upreti DK, Trivedi S, Shukla P. Antifungal activity of a common Himalayan foliose lichen Parmotrema tinctorum (Despr. ex Nyl.) Hale. Nat Sci 2011;9:167–171.
[15] Anjali DB, Mohabe S, Reddy AM, Nayaka S. Antimicrobial activities of 2-Propanol crude extract from lichen Parmotrema tinctorum (Despr. ex. Nyl.) Hale, collected from Eastern Ghats, India. Curr Res Environ Appl Mycol 2015;5:160–168.
[16] Vivek MN, Yashoda K, Manasa M, Prashith KTR. Antibacterial activity of three Parmotrema species from Western Ghats of Karnataka against clinical isolates of Burn and Dental caries. Sci Technol Arts Res J 2014;3:132–135.
[17] Poornima S, Ponmurugan P, Mythili Gnanamangai B, Nagarjun N, Ayyappadasan G. Evaluation of antimicrobial, antioxidant and anticancer activities of few macrolichens collected from Eastern Ghats of Tamil Nadu, India. Int Res J Pharm 2017;8:39–43.
[18] Ganesan A, Thangapandian M, Ponnusamy P, Sundararaj JP, Nayaka S. Antioxidant and antibacterial activity of parmeliod lichens from Shevaroy hills of Eastern Ghats, India. Int J Pharm Tech Res 2015;8:13–23.
[19] Ponmurugan P, Ayyappadasan G, Verma RS, Nayaka S. Survey, distribution pattern and elemental composition of lichens in Yercaud hills of Eastern Ghats in southern India. J Environ Biol 2016;37:407.
[20] Awasthi DD. A Compendium of the Macrolichens from India, Nepal and Sri Lank. Bishen Singh Mahendra Pal Sin; 2007.
[21] Orange A, James PW, White FJ. Microchemical Methods for the Identification of Lichens. Microchem Methods Identif Lichens 2001.
[22] Rajendran K, Poornima S, Ponnusamy P. Antimicrobial and Antiproliferative Activities of Depside Compound Isolated from the Mycobiont Culture of Parmotrema austrosinense (Zahlbr.) Hale. J Pure Appl Microbiol 2020;14:2525–41. https://doi.org/10.22207/JPAM.14.4.29.
[23] Culberson CF. Improved conditions and new data for identification of lichen products by standardized thin-layer chromatographic method. J Chromatogr A 1972;72:113–25. https://doi.org/10.1016/0021-9673(72)80013-X.
[24] OECD. Test No. 203: fish, acute toxicity test. OECD Publishing, Paris; 1992.
[25] Pittman J, Hylton A. Behavioral, endocrine, and neuronal alterations in zebrafish (Danio rerio) following sub-chronic coadministration of fluoxetine and ketamine. Pharmacol Biochem Behav 2015;139:158–62. https://doi.org/10.1016/j.pbb.2015.08.014.
[26] Cachat J, Stewart A, Grossman L, Gaikwad S, Kadri F, Chung KM, et al. Measuring behavioral and endocrine responses to novelty stress in adult zebrafish. Nat Protoc 2010;5:1786–1799.
[27] Krishnaraj C, Harper SL, Yun S-I. In Vivo toxicological assessment of biologically synthesized silver nanoparticles in adult Zebrafish (Danio rerio). J Hazard Mater 2016;301:480–91. https://doi.org/10.1016/j.jhazmat.2015.09.022.
[28] Lee PY, Costumbrado J, Hsu C-Y, Kim YH. Agarose Gel Electrophoresis for the Separation of DNA Fragments. JoVE J Vis Exp 2012:e3923. https://doi.org/10.3791/3923.
[29] Sambrook J, Russell DW. Alkaline agarose gel electrophoresis. CSH Protoc 2006.
[30] Wang D, Yang L, Wang J, Hu G, Liu Z, Yan D, et al. Behavioral and physiological effects of acute and chronic kava exposure in adult zebrafish. Neurotoxicol Teratol 2020;79:106881. https://doi.org/10.1016/j.ntt.2020.106881.
[31] Collier AD, Kalueff AV, Echevarria DJ. Zebrafish Models of Anxiety-Like Behaviors. In: Kalueff AV, editor. Rights Wrongs Zebrafish Behav. Phenotyping Zebrafish, Cham: Springer International Publishing; 2017, p. 45–72. https://doi.org/10.1007/978-3-319-33774-6_3.
[32] Maximino C, de Brito TM, da Silva Batista AW, Herculano AM, Morato S, Gouveia A. Measuring anxiety in zebrafish: A critical review. Behav Brain Res 2010;214:157–71. https://doi.org/10.1016/j.bbr.2010.05.031.
[33] Dutta HM. A composite approach for evaluation of the effects of pesticides on fish. Fish Morphol Horiz New Res Leban NH Sci Publ Pp 1996:249–269.
[34] Harvie EA, Huttenlocher A. Neutrophils in host defense: new insights from zebrafish. J Leukoc Biol 2015;98:523–37. https://doi.org/10.1189/jlb.4MR1114-524R.
[35] Matthias KA, Roche AM, Standish AJ, Shchepetov M, Weiser JN. Neutrophil-Toxin Interactions Promote Antigen Delivery and Mucosal Clearance of Streptococcus pneumoniae. J Immunol 2008;180:6246–54. https://doi.org/10.4049/jimmunol.180.9.6246.
[36] Woodfin A, Voisin M-B, Beyrau M, Colom B, Caille D, Diapouli F-M, et al. The junctional adhesion molecule JAM-C regulates polarized transendothelial migration of neutrophils in vivo. Nat Immunol 2011;12:761–769.
[37] Johnston IA, Bower NI, Macqueen DJ. Growth and the regulation of myotomal muscle mass in teleost fish. J Exp Biol 2011;214:1617–28. https://doi.org/10.1242/jeb.038620.
[38] Hong J-M, Kim JE, Min SK, Kim KH, Han SJ, Yim JH, et al. Anti-Inflammatory Effects of Antarctic Lichen Umbilicaria antarctica Methanol Extract in Lipopolysaccharide-Stimulated RAW 264.7 Macrophage Cells and Zebrafish Model. BioMed Res Int 2021;2021.
Published
07-10-2021
How to Cite
SHENBAGAM, M., M. MARIRAJ, R. KALIDOSS, M. ARUMUGAM DEEPA, and P. PONMURUGAN. “TOXICOLOGICAL AND BEHAVIOR ANALYSES OF A FOLIOSE LICHEN PARMOTREMA TINCTORUM (DELISE EX NYL.) HALE EXTRACT USING ZEBRAFISH (DANIO RERIO) MODEL”. Asian Journal of Pharmaceutical and Clinical Research, vol. 14, no. 10, Oct. 2021, pp. 27-32, doi:10.22159/ajpcr.2021.v14i10.42134.
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