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https://doi.org/10.53941/jmnp.2025.100005
Mizan, M., Miah, M. S., & Islam, M. K. Investigation of In-Vitro Antioxidant, Antimicrobial, Thrombolytic and In-Vivo Anti-Inflammatory and Antidepressant Potential of Holigarna Longifolia Leaf Extract. Journal of Medicinal Natural Products. 2025. doi: https://doi.org/10.53941/jmnp.2025.100005
Volume 2, Issue 1, 2025
Copyright (c) 2025 by the authors.
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Article

Investigation of In-Vitro Antioxidant, Antimicrobial, Thrombolytic and In-Vivo Anti-Inflammatory and Antidepressant Potential of Holigarna Longifolia Leaf Extract

Md Mizan 1,2,*, Md Shawon Miah 1 and Md Khairul Islam 1

1 Department of Pharmacy, Dhaka International University, Satarkul, Badda, Dhaka 1212, Bangladesh

2 Pharmacy Discipline, School of Life Science, Khulna University, Khulna 9208, Bangladesh

* Correspondence: hellotomizan@gmail.com; Tel.: +88-1746469198

Received: 11 January 2025; Revised: 9 February 2025; Accepted: 18 February 2025; Published: 20 February 2025

Abstract: Discovery of genus Holigarna longifolia and its traditional uses were obtained by living in forest or semi forest areas by close observations of the indigenous populations. The main aim of this study was to evaluate phytochemical screening, antioxidant and anti-microbial activity, thrombolytic activity, in vivo anti-inflammatory and neuropharmacological activity of H. longifolia leaf extract. In vitro antioxidant was evaluated by DPPH and ferric reducing power assay, Disc diffusion method was used to evaluate anti-microbial activity, thrombolytic activity was evaluated by clot lysis, determine the ability of the extracts to suppress inflammation was done using the formaldehyde-induced paw oedema test influence of the sedative activity of the drug on the forced swimming and tail flick-suspension tests were studied by using diazepam. The phytochemical screening of H. longifolia extract revealed the presence of reducing sugars, tannins, gums, steroids, glycosides, xanthoprotein and terpenoids. In DPPH the extract exhibited strong antioxidant with an IC50 of 0.877 µg/mL, comparable to ascorbic acid IC50 0.839 µg/mL and in ferric reducing power assay the extract showed significant IC50 value about 43.736 µg/mL. In anti-microbial tests, the highest inhibition, 19 mm was observed against Escherichia coli at 500 µg/disc, while Bacillus magaterium showed the lowest inhibition 9 mm at 250 µg/disc. In the MIC test, the extract leaves showed highest activity against E. coli (MIC = 4 mg/mL) and lowest against B. cereus, B. megaterium, and A. niger (MIC = 128 mg/mL). The extract demonstrates clot-lytic activity of 20.45 ± 1.42% versus 31.17 ± 0.65 for streptokinase. H. longifolia extract was determined to be safe in the acute toxicity trial, with no toxicity, mortality, or behavioral abnormalities in the mice. In the anti-inflammatory test, paw thickness was significantly reduced with inhibition rates of 54.22% by the dose of 250 mg/kg and 60.27% by 500 mg/kg) compared to 71.71% for the standard. Neuropharmacological tests revealed reduced immobility periods of 126.2 s in forced swimming and 145.8 s in tail suspension tests at dose of 200 mg/kg. Further studies will be needed to evaluate the full pharmacological potential H. longifolia.

Keywords:

phytochemicals anti-oxidant anti-microbial thrombolytic anti-inflammatory antidepressant

References

  1. Biswas, S.R.; Misbahuzzaman, K. Tree species diversity and regeneration traits of the dominant species in a dipterocarp forest in Bangladesh: Implications for conservation. Int. J. Biodivers. Sci. Manag. 2008, 4, 81–91.
  2. Singh, D.; Sharma, R.; Kumar, V. Phytochemical and pharmacological investigations of Holigarna longifolia: A review. J. Med. Plants Res. 2017, 11, 421–432.
  3. Rani, M.; Kumar, S.; Yadav, S. Phytochemical constituents and therapeutic potentials of Holigarna longifolia. Phytomedicine J. 2020, 29, 1–10.
  4. Patel, P.; Desai, S.; Mehta, M. Evaluation of antioxidant activity of Holigarna longifolia. Asian J. Pharm. Clin. Res. 2018, 11, 15–19.
  5. Kumar, P.; Yadav, R.; Mishra, P. Antimicrobial activity of Holigarna longifolia against clinical isolates. Int. J. Drug Dev. Res. 2019, 11, 45–50.
  6. Verma, P.; Meena, S.; Saini, N. Thrombolytic activity of Holigarna longifolia and its potential as an anticoagulant agent. Thromb. Res. 2021, 198, 35–42.
  7. Das, S.; Gupta, A.; Sharma, P. Neuropharmacological effects of Holigarna longifolia: A review. Neuropharmacol. Res. 2022, 52, 12–18.
  8. Soni, S.; Choudhury, M.; Gupta, R. Therapeutic potential of Holigarna longifolia: Antioxidant, antimicrobial, thrombolytic, and neuropharmacological activities. J. Herb. Med. 2023, 45, 107–116.
  9. Ghani, A.; Ahmad, S.B. Determination and identification of phytochemical properties of medicago sativa l.(alfalfa) leaf, stem and root extracts against various pathogens. Pak. J. Phytopathol. 2019, 31, 97–103.
  10. Baliyan, S.; Mukherjee, R.; Priyadarshini, A.; et al. Determination of Antioxidants by DPPH Radical Scavenging Activity and Quantitative Phytochemical Analysis of Ficus religiosa. Molecules 2022, 27, 1326.
  11. Govindarajan, R.; Rastogi, S.; Vijayakumar, M.; et al. Studies on the Antioxidant Activities of Desmodium gangeticum. Biol. Pharm. Bull. 2003, 26, 1424–1427.
  12. Benzie, I.F.F.; Strain, J.J. Ferric reducing/antioxidant power assay: Direct measure of total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration. Methods Enzymol. 1999, 299, 15–27.
  13. Fakruddin, M.; Mannan, K.S.B.; Mazumdar, R.M.; et al. Antibacterial, antifungal and antioxidant activities of the ethanol extract of the stem bark of Clausena heptaphylla. BMC Complement. Altern. Med. 2012, 12, 1–9.
  14. Behbahani, B.A.; Shahidi, F.; Yazdi, F.T.; et al. Antifungal effect of aqueous and ethanolic mangrove plant extract on pathogenic fungus “in vitro”. Int. J. Agron. Plant Prod. 2013, 4, 1652–1658.
  15. Prasad, S.; Kashyap, R.S.; Deopujari, J.Y.; et al. Development of an in vitro model to study clot lysis activity of thrombolytic drugs. Thromb. J. 2006, 4, 14.
  16. Watson, L.; Broderick, C.; Armon, M.P. Thrombolysis for acute deep vein thrombosis. Cochrane Database Syst. Rev. 2016, 11, 1465–1858.
  17. Manaharan, T.; Chakravarthi, S.; Radhakrishnan, A.K.; et al. In vivo toxicity evaluation of a standardized extract of Syzygium aqueum leaf. Toxicol. Rep. 2014, 1, 718–725.
  18. Garrido, G.; González, D.; Delporte, C.; et al. Analgesic and anti-inflammatory effects of Mangifera indica L. extract (Vimang). Phytother.Res. Int. J. Devoted Pharmacol. Toxicol. Eval. Nat. Prod. Deriv. 2001, 15, 18–21.
  19. Rd Bertin, M.; Jalfre, P.A. Behavioral despair in mice: A primary screening test for antidepressants. Arch. Int. Pharmacodyn. Ther. 1977, 229, 327–336.
  20. Steru, L.; Chermat, R.; Thierry, B.; et al. The tail suspension test: A new method for screening antidepressants in mice. Psychopharmacology 1985, 85, 367–370.
  21. Varadharajan, R.; Palani, S. Phytochemical screening, in-vitro antioxidant and cardioprotective potential of monochoria vaginalis on doxorubicin induced cardiotoxicity. J. Pharm. Biol. Sci. 2017, 5, 56.
  22. Agbafor, K.N.; Nwachukwu, N. Phytochemical Analysis and Antioxidant Property of Leaf Extracts of Vitex doniana and Mucuna pruriens. Biochem. Res. Int. 2011, 2011, 459839.
  23. Prakash, O.; Kumar, R.; Mishra, A.; et al. Artocarpus heterophyllus (Jackfruit): An overview. Pharmacogn. Rev. 2009, 3, 353.
  24. Hossain, A.; Hossain, A.; Mannan, S.J. Evaluation of antioxidant and analgesic activities of three medicinal plants. Pharmacogn. Res. 2019, 11, 248–253.
  25. Mukherjee, M.; Bhaskaran, N.; Srinath, R.; et al. Anti-ulcer and antioxidant activity of GutGardTM. Indian J. Exp. Biol. 2010, 48, 269–274.
  26. Ghalem, B.R.; Mohamed, B. Antibacterial activity of leaf essential oils of Eucalyptus globulus and Eucalyptus camaldulensis. Afr. J. Pharm. Pharmacol. 2008, 2, 211–215.
  27. Kotzekidou, P.; Giannakidis, P.; Boulamatsis, A. Antimicrobial activity of some plant extracts and essential oils against foodborne pathogens in vitro and on the fate of inoculated pathogens in chocolate. LWT—Food Sci. Technol. 2008, 41, 119–127.
  28. Yuan, J.; Yang, J.; Zhuang, Z.; et al. Thrombolytic effects of Douchi Fibrinolytic enzyme from Bacillus subtilis LD-8547 in vitro and in vivo. BMC Biotechnol. 2012, 12, 36.
  29. Afsar, T.; Khan, M.R.; Razak, S.; et al. Antipyretic, anti-inflammatory and analgesic activity of Acacia hydaspica R. Parker and its phytochemical analysis. BMC Complement. Altern. Med. 2015, 15, 1–12.
  30. Powar, P.; Gaikwad, D. GC-MS Analysis of bioactive compounds of Aegiceras corniculatum bark. Indian J. Plant Sci. 2016, 5, 13–17.
  31. Roome, T.; Razzak, A.; Ali, P.; et al. Therapeutic effect of Aegiceras corniculatum in chronic granulomatous inflammation and arthritis. J. Dow Univ. Health Sci. 2014, 8, 98–103.
  32. Subarnas, A.; Tadano, T.; Nakahata, N.; et al. A possible mechanism of antidepresant activity of beta-amyrin palmitate isolated from lobelia inflata leaves in the forced swimming test. Life Sci. 1993, 52, 289–296.
  33. Dhingra, D.; Sharma, A. Evaluation of antidepressant-like activity of glycyrrhizin in mice. Indian J. Pharmacol. 2005, 37, 390–394.