Amoebicidal activity of <i>Cassia angustifolia</i> extract and its effect on <i>Acanthamoeba triangularis</i> autophagy-related gene expression at the transcriptional level

Boonhok, Rachasak; Sangkanu, Suthinee; Norouzi, Roghayeh; Siyadatpanah, Abolghasem; Mirzaei, Farzaneh; Mitsuwan, Watcharapong; Charong, Nurdina; Wisessombat, Sueptrakool; Pereira, Maria de Lourdes; Rahmatullah, Mohammed; Wilairatana, Polrat; Wiart, Christophe; Tabo, Hazel Anne; Dolma, Karma G.; Nissapatorn, Veeranoot

doi:10.1017/s0031182021000718

Cited by 7 publications

(9 citation statements)

References 48 publications

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“…Expansion of the vacuole size may indicate a physiological change within the trophozoite in response to the stress induced by the P. harmala extract, and this change may be associated with basal autophagy in Acanthamoeba . The result was slightly different from our recent published data [ 56 ], that upon Cassia angustifolia leaf extract treatment, the percentage of the surviving parasite containing the enlarged vacuole was not increased. However, either P. harmala or C. angustifolia extract did not induce cyst formation on the surviving amoebae.…”

Section: Discussioncontrasting

confidence: 99%

“…On the other hand, the increase of Ac ATG16 may indicate other functions of this protein under the stress. In our recent study [ 56 ], the A. triangularis trophozoites treated with C. angustifolia extract and the ATG mRNA expression of ATG genes in surviving amoebae was analyzed in a similar manner. A similar pattern of the mRNA expression was observed.…”

Section: Discussionmentioning

confidence: 99%

“…A stock concentration of the plant extract was prepared at 100 mg/mL with 100% DMSO. The 96-well black plate (SPL Life Sciences, Seoul, Korea) was used for the amoebicidal activity testing and the final concentration of the extract was at 1 mg/mL [ 56 ]. For Acanthamoeba trophozoite harvesting, the parasite suspension was centrifuged, and the supernatant was discarded.…”

Section: Methodsmentioning

confidence: 99%

“…The tube was kept on ice along the process. After centrifugation, the culture medium was discarded and the parasite pellet was mixed with 500 µL of TRI reagent (Molecular Research Center, Cincinnati, OH, USA) to protect parasite RNA [ 56 ]. Total RNA extraction was done by RNA extraction kit (Vivantis Technologies, Selangor, Malaysia), and a 100 ng of total mRNA was converted to cDNA using the Viva cDNA synthesis kit (Vivantis Technologies, Selangor, Malaysia) following the manufacturer’s protocol.…”

Section: Methodsmentioning

confidence: 99%

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Peganum harmala Extract Has Antiamoebic Activity to Acanthamoeba triangularis Trophozoites and Changes Expression of Autophagy-Related Genes

et al. 2021

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Peganum harmala, a well-known medicinal plant, has been used for several therapeutic purposes as it contains numerous pharmacological active compounds. Our study reported an anti-parasitic activity of P. harmala seed extract against Acanthamoeba triangularis. The stress induced by the extract on the surviving trophozoites for Acanthamoeba encystation and vacuolization was examined by microscopy, and transcriptional expression of Acanthamoeba autophagy-related genes was investigated by quantitative PCR. Our results showed that the surviving trophozoites were not transformed into cysts, and the number of trophozoites with enlarged vacuoles were not significantly different from that of untreated control. Molecular analysis data demonstrated that the mRNA expression of tested AcATG genes, i.e., ATG3, ATG8b, and ATG16, was at a basal level along the treatment. However, upregulation of AcATG16 at 24 h post treatment was observed, which may indicate an autophagic activity of this protein in response to the stress. Altogether, these data revealed the anti-Acanthamoeba activity of P. harmala extract and indicated the association of autophagy mRNA expression and cyst formation under the extract stress, representing a promising plant for future drug development. However, further identification of an active compound and a study of autophagy at the protein level are needed.

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Section: Discussioncontrasting

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Peganum harmala Extract Has Antiamoebic Activity to Acanthamoeba triangularis Trophozoites and Changes Expression of Autophagy-Related Genes

et al. 2021

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“…Trophozoites with vacuoles, regardless of their size, as well as the trophozoites containing enlarged vacuoles, were examined. An enlarged vacuole (EV) was defined as a vacuole with a diameter of at least 5 µm, and the trophozoite containing at least one EV was counted as 1 ( Boonhok et al, 2021b ). The experiment was performed with three independent experiments.…”

Section: Methodsmentioning

confidence: 99%

Curcumin effect on Acanthamoeba triangularis encystation under nutrient starvation

Boonhok

Sangkanu

Phumjan

et al. 2022

PeerJ

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Background Curcumin is an active compound derived from turmeric, Curcuma longa, and is known for its benefits to human health. The amoebicidal activity of curcumin against Acanthamoeba triangularis was recently discovered. However, a physiological change of intracellular pathways related to A. triangularis encystation mechanism, including autophagy in the surviving amoeba after curcumin treatment, has never been reported. This study aims to investigate the effect of curcumin on the survival of A. triangularis under nutrient starvation and nutrient-rich condition, as well as to evaluate the A. triangularis encystation and a physiological change of Acanthamoeba autophagy at the mRNA level. Methods In this study, A. triangularis amoebas were treated with a sublethal dose of curcumin under nutrient starvation and nutrient-rich condition and the surviving amoebas was investigated. Cysts formation and vacuolization were examined by microscopy and transcriptional expression of autophagy-related genes and other encystation-related genes were evaluated by real-time PCR. Results A. triangularis cysts were formed under nutrient starvation. However, in the presence of the autophagy inhibitor, 3-methyladenine (3-MA), the percentage of cysts was significantly reduced. Interestingly, in the presence of curcumin, most of the parasites remained in the trophozoite stage in both the starvation and nutrient-rich condition. In vacuolization analysis, the percentage of amoebas with enlarged vacuole was increased upon starvation. However, the percentage was significantly declined in the presence of curcumin and 3-MA. Molecular analysis of A. triangularis autophagy-related (ATG) genes showed that the mRNA expression of the ATG genes, ATG3, ATG8b, ATG12, ATG16, under the starvation with curcumin was at a basal level along the treatment. The results were similar to those of the curcumin-treated amoebas under a nutrient-rich condition, except AcATG16 which increased later. On the other hand, mRNA expression of encystation-related genes, cellulose synthase and serine proteinase, remained unchanged during the first 18 h, but significantly increased at 24 h post treatment. Conclusion Curcumin inhibits cyst formation in surviving trophozoites, which may result from its effect on mRNA expression of key Acanthamoeba ATG-related genes. However, further investigation into the mechanism of curcumin in A. triangularis trophozoites arrest and its association with autophagy or other encystation-related pathways is needed to support the future use of curcumin.

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Senna (Cassia angustifolia Vahl.): A comprehensive review of ethnopharmacology and phytochemistry

Thaker,

Patoliya,

Rabadiya

et al. 2023

Pharmacological Research - Natural Products

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Amoebicidal activity of Cassia angustifolia extract and its effect on Acanthamoeba triangularis autophagy-related gene expression at the transcriptional level

Abstract: Abstract

Cited by 7 publications

References 48 publications

Peganum harmala Extract Has Antiamoebic Activity to Acanthamoeba triangularis Trophozoites and Changes Expression of Autophagy-Related Genes

Peganum harmala Extract Has Antiamoebic Activity to Acanthamoeba triangularis Trophozoites and Changes Expression of Autophagy-Related Genes

Curcumin effect on Acanthamoeba triangularis encystation under nutrient starvation

Senna (Cassia angustifolia Vahl.): A comprehensive review of ethnopharmacology and phytochemistry

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