Induction of Apoptosis by Alcoholic Extract of Combination Verbascum thapsus and Ginger officinale on Iranian Isolate of Trichomonas vaginalis
AbstractBackground: The protozoan Trichomonas vaginalis is a sexually transmitted disease (STD). Metronidazole is a chosen drug for the treatment. This study evaluated the anti trichomonal activity of alcoholic extracts of combination Verbascum thapsus and Ginger officinale.Methods: This experimental study was conducted in the Parasitology Laboratory, Kashan University of Medical Sciences, Kashan, Iran in 2015, on 23 women with suspected trichomoniasis referring to Kashan clinical centers. Medium TYI-S-33 was used for culture of three T. vaginalis isolates. Different concentrations (25, 50, 100, 200, 400, 800 µg/ml) of V. thapsus and G. officinale ethanol extract added to Trichomonas trophozoites in 48-well plates and metronidazole considered as positive control and the negative control was TYI-S33 containing Trichomonas trophozoites without any drug. In all of mentioned groups, trophozoites number counted 12, 24, 48 h after culture. Results were analyzed using ANOVA statistical test, to evaluate the toxicity of extract, measured by MTT assay. Induced apoptosis of T. vaginalis after treatment with different concentrations of extract was determined by Flow Cytometry. Results: IC50 of alcoholic extract of combination V. thapsus and G. officinale and metronidazole after 24h was 73.80 µg/ml and 0.0326 µg/ml, respectively. The toxicity percentage of 25-800 μg/ml concentrations of this combination were between 0.2-1.98. In different concentrations of extract (25,50,100,200 and 400 µg/ml) apoptosis percent after 48h was 18.97 to 77.19 and necrosis percent was calculated 1.35, 3.18, 3.10, 1.16 and 4.09, respectively.Conclusion: Alcoholic extract of combination V. thapsus and G. officinale induces programmed death in T. vaginalis. Due to no toxicity on macrophages, it can be examined in vivo studies.
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