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

Shamseddin J, Akhlaghi L, Razmjou E, Shojaee S, Monavari SH, Tajik N, Ebrahimi SA, Meamar AR - Iran J Parasitol (2015 Apr-Jun)

Bottom Line: The results of MTT indicated that CLA could decrease the proliferation of tachyzoites of both strains in HeLa cells.Conjugated linoleic acid has anti-toxoplasmacidal activity on tachyzoites of T. gondii.Therefore, we recommended further studies on this component in order to achieve a new drug against the parasite.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Parasitology and Mycology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.

ABSTRACT

Background: The aim of this study was to evaluate the effects of conjugated linoleic 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 cytometry using Annexin V and propidium iodide (PI) double staining. The statistical analysis 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 tachyzoites of T. gondii. Therefore, we recommended further studies on this component in order to achieve a new drug against the parasite.

No MeSH data available.


Related in: MedlinePlus

Flow cytometry results. (A) Maximum apoptosis rates at 24 (I), 48 (II), and 72 (III) h post-treatment of RH strain Toxoplasma tachyzoites with CLA. (B) The highest level of apoptosis was with 200 μM CLA after 24 h post-treatment of Tehran strain (I). The highest apoptosis at 10 μM CLA was after 48 h (II). Late apoptosis at 250 μM CLA after 72 h is shown in Q2 of graph (III). The right lower quadrant of each chart shows cells stained with Annexin V
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Figure 2: Flow cytometry results. (A) Maximum apoptosis rates at 24 (I), 48 (II), and 72 (III) h post-treatment of RH strain Toxoplasma tachyzoites with CLA. (B) The highest level of apoptosis was with 200 μM CLA after 24 h post-treatment of Tehran strain (I). The highest apoptosis at 10 μM CLA was after 48 h (II). Late apoptosis at 250 μM CLA after 72 h is shown in Q2 of graph (III). The right lower quadrant of each chart shows cells stained with Annexin V

Mentions: Flow cytometry assay showed that less than 5 % of tachyzoites dead in the control wells (0 μM CLA). After 24 h treatment of the RH strain tachyzoites, maximum early apoptosis (PS exposure detected by Annex V staining) (39.3%) was observed at 100 μM CLA. At higher concentrations, apoptosis decreased (approximately 20% at 150 to 250 μM). Late apoptosis (disintegration of other organelles of parasites detected by PI staining) was highest with the 50 μM treatment (36.1%) and was not considerable at other exposure times and concentrations in the RH strain (Table 1). After forty-eight hours exposure to CLA at 200 μM, apoptosis increased. The apoptosis rate was 9.8% at 30 μM CLA and increased to a maximum cell death of 54.0% at the concentration of 200 μM. After 72 h, the maximum rate of apoptosis (51.4%) was seen at 250 μM CLA (Fig. 2A).


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

Shamseddin J, Akhlaghi L, Razmjou E, Shojaee S, Monavari SH, Tajik N, Ebrahimi SA, Meamar AR - Iran J Parasitol (2015 Apr-Jun)

Flow cytometry results. (A) Maximum apoptosis rates at 24 (I), 48 (II), and 72 (III) h post-treatment of RH strain Toxoplasma tachyzoites with CLA. (B) The highest level of apoptosis was with 200 μM CLA after 24 h post-treatment of Tehran strain (I). The highest apoptosis at 10 μM CLA was after 48 h (II). Late apoptosis at 250 μM CLA after 72 h is shown in Q2 of graph (III). The right lower quadrant of each chart shows cells stained with Annexin V
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Related In: Results  -  Collection

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Figure 2: Flow cytometry results. (A) Maximum apoptosis rates at 24 (I), 48 (II), and 72 (III) h post-treatment of RH strain Toxoplasma tachyzoites with CLA. (B) The highest level of apoptosis was with 200 μM CLA after 24 h post-treatment of Tehran strain (I). The highest apoptosis at 10 μM CLA was after 48 h (II). Late apoptosis at 250 μM CLA after 72 h is shown in Q2 of graph (III). The right lower quadrant of each chart shows cells stained with Annexin V
Mentions: Flow cytometry assay showed that less than 5 % of tachyzoites dead in the control wells (0 μM CLA). After 24 h treatment of the RH strain tachyzoites, maximum early apoptosis (PS exposure detected by Annex V staining) (39.3%) was observed at 100 μM CLA. At higher concentrations, apoptosis decreased (approximately 20% at 150 to 250 μM). Late apoptosis (disintegration of other organelles of parasites detected by PI staining) was highest with the 50 μM treatment (36.1%) and was not considerable at other exposure times and concentrations in the RH strain (Table 1). After forty-eight hours exposure to CLA at 200 μM, apoptosis increased. The apoptosis rate was 9.8% at 30 μM CLA and increased to a maximum cell death of 54.0% at the concentration of 200 μM. After 72 h, the maximum rate of apoptosis (51.4%) was seen at 250 μM CLA (Fig. 2A).

Bottom Line: The results of MTT indicated that CLA could decrease the proliferation of tachyzoites of both strains in HeLa cells.Conjugated linoleic acid has anti-toxoplasmacidal activity on tachyzoites of T. gondii.Therefore, we recommended further studies on this component in order to achieve a new drug against the parasite.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Parasitology and Mycology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.

ABSTRACT

Background: The aim of this study was to evaluate the effects of conjugated linoleic 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 cytometry using Annexin V and propidium iodide (PI) double staining. The statistical analysis 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 tachyzoites of T. gondii. Therefore, we recommended further studies on this component in order to achieve a new drug against the parasite.

No MeSH data available.


Related in: MedlinePlus