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Original Article

Sterculic Acid and Its Analogues Are Potent Inhibitors of Toxoplasma gondii

Pan Hao1, Intisar Q. M. Alaraj2, Juma’a R. Al Dulayymi2, Mark S. Baird2, Jing Liu1, Qun Liu1,*
The Korean Journal of Parasitology 2016;54(2):139-145.
Published online: April 30, 2016

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*Corresponding author (qunliu@cau.edu.cn)
• Received: December 2, 2015   • Revised: March 10, 2016   • Accepted: March 13, 2016

© 2016, Korean Society for Parasitology and Tropical Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Sterculic Acid and Its Analogues Are Potent Inhibitors of Toxoplasma gondii
Korean J Parasitol. 2016;54(2):139-145.   Published online April 30, 2016
DOI: https://doi.org/10.3347/kjp.2016.54.2.139
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Sterculic Acid and Its Analogues Are Potent Inhibitors of Toxoplasma gondii
Korean J Parasitol. 2016;54(2):139-145.   Published online April 30, 2016
DOI: https://doi.org/10.3347/kjp.2016.54.2.139
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Sterculic Acid and Its Analogues Are Potent Inhibitors of Toxoplasma gondii
Image Image Image Image
Fig. 1. Molecular structures of test compounds. Sterculic acid was coded as INT16. The methyl esters INT13, INT14, and INT15 are different in the length of carbon chain. INT21 has the same length of carbon chain with sterculic acid but contains a methoxy moiety.
Fig. 2. Sterculic acid and methyl esters inhibited the egress of tachyzoites from Vero cells. Representative pictures were presented to show that most of host cells remained intact after incubation with INT13 (A), INT14 (B), INT15 (C), INT16 (D), or INT21 (E) for 36 hr, with a small number of released tachyzoites, compared with numerous tachyzoites and a smaller number of intact host cells in the negative control group (F). Scale bar=100 μM. Extracellular tachyzoites are marked in red circles. (G) The number of released tachyzoites in the culture incubated with the test compounds was decreased relative to the DMSO control. Extracellular tachyzoites were counted in 6 random fields under the microscope per group. Data are presented as mean±SD. **P<0.01.
Fig. 3. Sterculic acid and all methyl ester analogues inhibit tachyzoite proliferation. Freshly released parasites were used to infect HFF cell monolayers. (A) Invaded parasites were allowed to replicate for 18 hr in the culture medium containing the test compounds, and the number of vacuoles containing 1, 2, 4, or 8 parasites was counted under a fluorescence microscope after anti-T. gondii staining. (B) Representative fluorescent microscopic images were presented to show that many vacuoles contained 8 tachyzoites in DMSO control groups, (C) while 4 or 2 tachyzoites were often observed in a vacuole in INT21-treated groups after culture for 18 hr. The magnified images of the areas marked with white boxes in (B) and (C) were presented at the bottom in order. Data are presented as mean±SD from 3 independent experiments each in triplicate. **P<0.01.
Fig. 4. Relative cell viability was calculated from the MTS assay. The cell viability of 0.1% DMSO was regarded as 100%. (A) Sterculic acid was cytotoxic to Vero cells at high concentrations while the methyl esters displayed no cytotoxicity. (B) All test compounds inhibited T. gondii growth as shown by increased viability of Vero cells infected with T. gondii.

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Sterculic Acid and Its Analogues Are Potent Inhibitors of Toxoplasma gondii
Compound code ECa50 (μM) CCb50 (μM) SIc
Sterculic acid (INT16) 36.2 ± 5.1 215.2 ± 15.8 5.9
INT13 356.9 ± 14.1 > 1,000 > 2.8
INT14 428.4 ± 15.7 > 1,000 > 2.3
INT15 306.4 ± 18.8 > 1,000 > 3.3
INT21 248.0 ± 17.9 > 1,000 > 4.0
Table 1. In vitro anti-T. gondii activity of sterculic acid and its methyl ester analogues

Results are presented as mean±SD from 3 independent experiments with triplicate wells in each experiment.

EC50, half maximal effective concentration. Concentration required to inhibit T. gondii-induced cytopathic effect by 50%.

CC50, half maximal cytotoxic concentration. Concentration required to reduce Vero cell growth by 50%.

Selectivity index=CC50/EC50

Table 1.

PHD : Parasites, Hosts and Diseases

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