Autophagic cell death induction in a clinical isolate of Acanthamoeba sp. treated  with methanolic extracts of Pereskia bleo Kuh

Razifah Mohd Razali; Fatin Amira Abdullah; Nur Nabilah Mohd Basri; Syed Ahmad Tajudin Tuan Johari; Thivyan Manisekaran; Fatimah Hashim

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Autophagic cell death induction in a clinical isolate of Acanthamoeba sp. treated with methanolic extracts of Pereskia bleo Kuh

Author: Razifah Mohd Razali ^{1
,

2} ; Fatin Amira Abdullah ¹ ; Nur Nabilah Mohd Basri ¹ ; Syed Ahmad Tajudin Tuan Johari ³ ; Thivyan Manisekaran ¹ ; Fatimah Hashim ^{1
,

2}
Author Information

1. Faculty of Science and Marine Environment, Universiti Malaysia Terengganu, 21030 Kuala Nerus, Terengganu, Malaysia.&
2. Biological Security and Sustainability Research Group, Faculty of Science and Marine Environment, Universiti Malaysia Terengganu, 21030 Kuala Nerus, Terengganu, Malaysia.
3. Centre for Research in Infectious Diseases and Biotechnology, Faculty of Medicine, Universiti Sultan Zainal Abidin, Medical Campus. 20400 Kuala Terengganu, Terengganu, Malaysia.
Publication Type:Journal Article
Keywords: Acanthamoeba keratitis; Annexin V; antioxidant; autophagy; β-carotene
From:Malaysian Journal of Microbiology 2024;20(no.2):224-236
CountryMalaysia
Language:English
Abstract: Aims:This study was aimed to investigate the effects of methanol extracts from various parts of the Pereskia bleo Kuhn plant on Acanthamoeba sp. The antioxidant levels of each extract from different plant parts were measured after the extraction process. These extracts were then exposed to Acanthamoeba sp. to assess dose-response, IC50 values, changes in cell morphology, internal cell activity and apoptosis based on alterations in phospholipids.
Methodology and results:The total phenolic content, carotenoid estimation and antioxidant activity of the leaves, flowers and fruits of P. bleo were measured based on 1,1-diphenyl-2-picrylhydrazyl radicals (DPPH) assay. Its anti-amoebic properties were tested using 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay for the IC50 determination. The morphological and biochemical changes in the Acanthamoeba sp. were observed under light and fluorescence microscopy using the acridine orange and propidium iodide double staining (AO/PI). The IC50 values of P. bleo leaves, flowers and fruits methanolic extracts were 5.884%, 0.1646% and 20.69%, respectively. Morphological observation displayed shortened acanthapodia with darkened cytoplasms. AO/PI-stained Acanthamoeba sp. cells appear with orange-fluorescent organelles in their green cytoplasm, indicating autophagic cell deaths. Apoptotic and necrotic Acanthamoeba sp. cells were absent based on Annexin V labelling.
Conclusion, significance and impact of study: :This study confirmed that the methanolic crude extracts of P. bleo exhibit high cytotoxic potential towards Acanthamoeba sp. trophozoites by inducing an autophagic mode of cell death.
Full text:20.2024my0027.pdf