Vet World   Vol.17   December-2024  Article - 13 

Research Article

Veterinary World, 17(12): 2829-2845

https://doi.org/10.14202/vetworld.2024.2829-2845

Anti-adherent effects of Rhizophora apiculata bark and leaf extracts and computational prediction of the effects of its compound on β-tubulin interaction in Acanthamoeba triangularis genotype 4

Siriphorn Chimplee1, Imran Sama-ae2, Suthinee Sangkanu3, Watcharapong Mitsuwan4, Julalak Chuprom1, Rachasak Boonhok2, Dhrubo Ahmed Khan5, Partha Biswas5,6, Md Nazmul Hasan5, Hazel Anne Tabo7, Cristina C. Salibay7, Polrat Wilairatana8, Maria L. Pereira9, Muhammad Nawaz10, Ragini Bodade11, Shanmuga S. Sundar12, Alok K. Paul13, and Veeranoot Nissapatorn3
1. General Education Department, School of Languages and General Education, Walailak University, Nakhon Si Thammarat, 80160, Thailand.
2. Department of Medical Technology, School of Allied Health Sciences and Center of Excellence Research for Melioidosis and Microorganisms (CERMM), Walailak University, Nakhon Si Thammarat, 80160, Thailand.
3. School of Allied Health Sciences, Southeast Asia Water Team (SEA Water Team) and World Union for Herbal Drug Discovery (WUHeDD), Walailak University, Nakhon Si Thammarat, 80160, Thailand.
4. Akkhraratchakumari Veterinary College, Walailak University, Nakhon Si Thammarat, 80160, Thailand.
5. Laboratory of Pharmaceutical Biotechnology and Bioinformatics, Department of Genetic Engineering and Biotechnology, Jashore University of Science and Technology, Jashore-7408, Bangladesh.
6. ABEx Bio-Research Center, East Azampur, Dhaka-1230, Bangladesh.
7. Department of Biological Sciences, College of Science, De La Salle University-Dasmarinas, Cavite, Philippines.
8. Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
9. Department of Medical Sciences and CICECO-Aveiro Institute of Materials, University of Aveiro, Aveiro, Portugal.
10. Department of Nano-Medicine Research, Institute for Research and Medical Consultations, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia.
11. Life Science Division, Institute of Advanced Study in Science and Technology (IASST), Vigyan Path, Paschim Boragaon, Garchuk, Guwahati, Assam, India.
12. Department of Biotechnology, Aarupadai Veedu Institute of Technology, Vinayaka Mission’s Research Foundation, Paiyanoor, Chennai, Tamil Nadu, India.
13. School of Pharmacy and Pharmacology, University of Tasmania, Hobart, TAS 7001, Australia. 

Background and Aim: Acanthamoeba, an opportunistic protozoan, exists widely in natural sources and can cause infections in humans and animals. The absence of effective monotherapy after the initial infection leads to chronic disease and recurrence. Tubulin protein is a vital target for design-targeted drug discovery. Anti-tubulin drugs are also used to treat Acanthamoeba infections, although resistance to these drugs has been observed. Therefore, it is necessary to identify a new targeted drug for Acanthamoeba infections. Therefore, this study aimed to assess the in vitro activity of ethanol extracts of Rhizophora apiculata extracts (RAE) against Acanthamoeba spp. and to predict its chemical compound on β-tubulin interaction. 

Materials and Methods: In this study, anti-Acanthamoeba activity with minimal inhibitory concentration (MIC) and minimal parasiticidal concentration (MPC) determination of ethanolic RAE from leaves, blossoms, buds, branches, and barks was tested on four Acanthamoeba trophozoites and cysts: Acanthamoeba triangularis WU 19001, Acanthamoeba polyphaga American Type Culture Collection (ATCC) 30461, Acanthamoeba castellanii ATCC 50739, and A. castellanii ATCC 30010. The inhibitory effect on adherence was determined by the ability of Acanthamoeba adherence on 96-well plates, and its adhesive acanthopodia structure was evaluated using scanning electron microscopy analysis. In addition, the minimum cytotoxic concentrations (MCC) of R. apiculata leaf extract (RALE) and bark extract (RABE) were evaluated on Vero and HaCaT cell lines using the MTT assay. Phytochemical compounds from RALE and RABE were also analyzed by gas chromatography-mass spectrometry (GC-MS). Molecular docking and molecular dynamic analysis predicted the binding sites of chemicals in extracts and β-tubulin protein.

Results: The results revealed that A. triangularis and A. polyphaga trophozoites had the highest inhibition at 90% at a MIC of 8 mg/mL after treatment with RALE and RABE, respectively, at 24 h. Those MPC values were exhibited at 16 mg/mL against A. triangularis trophozoites. In addition, both extracts inhibited the adhesive properties of all Acanthamoeba approximately 80%–90% at 4 mg/ mL, as well as adherent structural acanthopodia loss. MCC was 0.25 mg/mL, provided to be harmless to mammalian cells. GC-MS analysis supported that 8 and 11 major phytochemicals were from RABE and RALE, respectively. Molecular docking and molecular dynamics demonstrated that Acanthamoeba-β-tubulin exhibited potent root-mean-square deviation, root mean square fluctuation, and binding free energy values with clionasterol (from RABE and RALE) and stigmasterol (from RALE). Based on our results, ethanolic RABE and RALE exhibited anti-Acanthamoeba activity in reducing adhesion. In silico showed that promising clionasterol and stigmasterol interacted with a targeting β-tubulin. 

Conclusion: The RABE and RALE exhibited a potential adherent effect on A. triangularis, low toxicity, and the clionasterol and stigmasterol in RABE and RALE predicted to interact the targeted β-tubulin. These agents may be used as alternative therapeutic agents in the management of disease using a sustainable one-heath approach. 

Keywords: Acanthamoeba triangularis, anti-adherent activity, molecular docking, molecular dynamic simulation, Rhizophora, tubulin.

How to cite this article: Chimplee S, Sama-ae I, Sangkanu S, Mitsuwan W, Chuprom J, Boonhok R, Khan DA, Biswas P, Hasan MN, Tabo HA, Salibay CC, Wilairatana P, Pereira ML, Nawaz M, Bodade R, Sundar SS, Paul AK, and Nissapatorn V (2024) Anti-adherent effects of Rhizophora apiculata bark and leaf extracts and computational prediction of the effects of its compound on β-tubulin interaction in Acanthamoeba triangularis genotype 4, Veterinary World, 17(12): 2829–2845.

Received: 2024-06-01    Accepted: 2024-11-12    Published online: 2024-12-18

Corresponding author: Veeranoot Nissapatorn    E-mail: nissapat@gmail.com

DOI: 10.14202/vetworld.2024.2829-2845

Copyright: Chimplee, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/ by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/ publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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