Original Article

Conjugated Linoleic Acid Stimulates Apoptosis in RH and Tehran Strains of Toxoplasma gondii, in Vitro


Background:The aim of this study was to evaluate the effects of conjugated lino-leic acid (CLA) on apoptosis of tachyzoites of T. gondii, RH strain (type I) and the cyst-forming Tehran strain (type II) in vitro.
Methods:Toxoplasma strains were injected into the peritoneal cavity of BALB/c mice. The Tehran strain forms cysts in the brain of mice. Bradyzoites within the cysts are reactivated to proliferative tachyzoites, by dexamethasone. Tachyzoites were aspirated from the peritoneum of infected mice, and the percentage of viable parasites was estimated with trypan blue staining. Tachyzoites were inoculated into HeLa cells cultivated in DMEM medium. Different concentrations of CLA were evaluated on T. gondii in HeLa cells by the tetrazolium (MTT) colorimetric assay. Differentiation between apoptosis and cell death was determined by flow cytome-try using Annexin V and propidium iodide (PI) double staining. The statistical anal-ysis performed by GraphPad Prism version 6.00.
Results:CLA induces apoptosis in virulent (RH) and avirulent (Tehran) strains of T. gondii. The results of MTT indicated that CLA could decrease the proliferation of tachyzoites of both strains in HeLa cells.
Conclusion:Conjugated linoleic acid has anti-toxoplasmacidal activity on tachyzo-ites of T. gondii. Therefore, we recommended further studies on this component in order to achieve a new drug against the parasite.

Dubey JP. Toxoplasmosis of animals and humans. Second ed. Maryland: CRC Press; 2010.

Derouin F, Pelloux H. Prevention of toxoplasmosis in transplant patients. Clin Microbiol Infect. 2008; 14(12):1089–101.

Jones-Brando L, D'Angelo J, Posner GH, Yolken R. In vitro inhibition of Toxoplasma gondii by four new derivatives of artemisinin. Antimicrob Agents Chemother. 2006; 50(12):4206–8.

Niezgoda N, Wawrzeńczyk C. An efficient method for enzymatic purification of cis-9, trans-11 isomer of conjugated linoleic acid. J Mol Catal B Enzym. 2014; 100:40–8.

Stanimirovic M, Petrujkic B, Delic N, Djelic N, Stevanovic J, Stanimirovic Z. Dietary conjugated linoleic acid influences the content of stearinic acid in porcine adipose tissue. Veterinarni Medicina. 2012; 57(2):92–100.

Kumaratilake LM, Robinson BS, Ferrante A, Poulos A. Antimalarial properties of n-3 and n-6 polyunsaturated fatty acids: in vitro effects on Plasmodium falciparum and in vivo effects on P. berghei. J Clin Invest. 1992; 89(3):961–7.

Melariri P, Campbell W, Etusim P, Smith P. In vitro and in vivo antimalarial activity of linolenic and linoleic acids and their methyl esters. Adv Stud Biol. 2012; 4(7):333–49.

Shareck J, Nantel A, Belhumeur P. Conjugated linoleic acid inhibits hyphal growth in Candida albicans by modulating Ras1p cellular levels and downregulating TEC1 expression. Eukaryot Cell. 2011; 10(4):565–77.

Tanmahasamut P, Liu J, Hendry LB, Sidell N. Conjugated linoleic acid blocks estrogen signaling in human breast cancer cells. J Nutr. 2004; 134(3):674–80.

Ghorbani M, Samii AH. Toxoplasmic lymphadenitis in Iran. J Trop Med Hyg. 1973; 76(7):158–60.

Saeij JP, Boyle JP, Grigg ME, Arrizabalaga G, Boothroyd JC. Bioluminescence imaging of Toxoplasma gondii infection in living mice reveals dramatic differences between strains. Infect Immun. 2005; 73(2):695–702.

Ashburn D, Evans R, Chatterton JM, Joss AW, Ho-Yen DO. Toxoplasma dye test using cell culture derived tachyzoites. J Clin Pathol. 2000; 53(8):630–3.

van Meerloo J, Kaspers GL, Cloos J. Cell Sensitivity Assays: The MTT Assay. In: Cree IA, editor. Cancer Cell Culture: Humana Press; 2011. p. 237–45.

Zhang G, Gurtu V, Kain SR, Yan G. Early detection of apoptosis using a fluorescent conjugate of annexin V. BioTechniques. 1997; 23(3):525–31.

Laane E, Tamm KP, Buentke E, Ito K, Khahariza P, Oscarsson J, et al. Cell death induced by dexamethasone in lymphoid leukemia is mediated through initiation of autophagy. Cell Death Differ. 2009; 16(7):1018–29.

Calder PC. Polyunsaturated fatty acids, inflammation, and immunity. Lipids. 2001; 36(9):1007–24.

Henderson WR, Jr., Rashed M, Yong EC, Fritsche TR, Chiang GK. Toxoplasma gondii stimulates the release of 13- and 9-hydroxyoctadecadienoic acids by human platelets. Biochemistry. 1992; 31(23):5356–62.

Henderson WR, Chi EY. Cytotoxic activity of 13-hydroxyoctadecadienoic acid against Toxoplasma gondii. Parasitology. 1992; 105(03): 343–7.

Doliwa C, Xia D, Escotte-Binet S, Newsham EL, Sanya J S, Aubert D, et al. Identification of differentially expressed proteins in sulfadiazine resistant and sensitive strains of Toxoplasma gondii using difference-gel electrophoresis (DIGE). Int J Parasitol Drugs Drug Resist. 2013; 3:35–44.

Sarkar FH, Li Y. Markers of apoptosis. Breast Cancer Research Protocols: Springer; 2006. p. 147–60.

Patel VA, Longacre A, Hsiao K, Fan H, Meng F, Mitchell JE, et al. Apoptotic cells, at all stages of the death process, trigger characteristic signaling events that are divergent from and dominant over those triggered by necrotic cells: Implications for the delayed clearance model of autoimmunity. J Biol Chem. 2006; 281(8):4663–70.

IssueVol 10 No 2 (2015) QRcode
SectionOriginal Article(s)
Apoptosis Conjugated linoleic acid (CLA) RH strain Toxoplasma gondii

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How to Cite
Shamseddin J, Akhlaghi L, Razmjou E, Shojaee S, Monavari SHR, Tajik N, Ebrahimi SA, Meamar AR. Conjugated Linoleic Acid Stimulates Apoptosis in RH and Tehran Strains of Toxoplasma gondii, in Vitro. Iran J Parasitol. 10(2):238-244.