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. 2014 Apr-Jun;9(2):202-8.

Triclabendazole (Anthelmintic Drug) Effects on the Excretory- Secretory Proteome of Fasciola hepatica in Two Dimension Electrophoresis Gel

Ashkan Faridi  1 , Ali Farahnak  1 , Taghi Golmohammadi  2 , Mohammadreza Eshraghian  3 , Yosef Sharifi  1 , Mohammadbagher Molaei Rad  1
Affiliations

Triclabendazole (Anthelmintic Drug) Effects on the Excretory- Secretory Proteome of Fasciola hepatica in Two Dimension Electrophoresis Gel

Ashkan Faridi et al. Iran J Parasitol. 2014 Apr-Jun.

Abstract

Background: The aim of this study was to evaluate the protein spots of excretory - secretory products of Fasciola hepatica using two dimension electrophoresis method in the presence and absence of triclabendazole drug which can be considered to detect the target protein of the drug.

Methods: F. hepatica parasites were collected from infected cattle livers, divided in two groups and cultivated in RPMI 1640 medium. First group was treated with triclabendazole (TCBZ) and second group considered as control. The excretory-secretory (ES) products of each group were separated and total protein determined by Bradford method. To provide proteome spots, the ES proteins were precipitated and two dimension electrophoresis (2-DE) gel prepared. Protein amounts of two groups were compared using the statistical t-test and protein spots from 2-DE in test and control groups were also statistically analyzed. The protein spots of gels were identified by using protein database.

Results: The t-test showed a significant increase of total proteins in treated group (P<0.5). The protein spots count in the control group was less than test group however statistically not significant (p>0.05). Cathepsin L- protein (MW 36.7 pH 5.34), 14-3-3 epsilon 2 isoform (MW 28.2 pH 5.36), Cathepsin L1D (MW 36.5 pH 5.8) and Cathepsin L1D (MW 36.6 pH 6.26) were identified in test group.

Conclusion: It seems that, these results can be considered to determine the proteins which the drug acts as a target on them.

Keywords: Fasciola hepatica; Triclabendazole Proteome; Two - dimensional electrophoresis (2-DE).

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Figures

Fig. 1
Fig. 1
The Protein spots of Fasciola hepatica ES products, control sample (C8), in 2-DE gel. Spot number 1 (MW 53.2 pH 6.6), Spot number 6 (MW 30.0 pH 5.9), Spot number 8 (MW 31.6 pH 6.4), Spot number 9 (MW 30.0 pH 6.4), Spot number 12 (MW 33.6 pH 6.7), Spot number 13 (MW 33.1 pH 7.0), Spot number 14 (MW 31.6 pH 7.1), Spot number 15 (MW 33.1 pH 7.6), Spot number 16 (MW 31.6 pH 7.6), Spot number 18 (MW 32.0 pH 8.6), M; protein marker
Fig. 2
Fig. 2
The Protein spots of Fasciola hepatica ES products, test sample (T8), in 2-DE gel. Spot number 1 (MW 53.2 pH 6.6), Spot number 5 (MW 31.6 pH 5.9), Spot number 6 (MW 30.0 pH 5.9), Spot number 8 (MW 31.6 pH 6.4), Spot number 9 (MW 30.0 pH 6.4), Spot number 11 (MW 33.1 pH 6.6), Spot number 12 (MW 33.6 pH 6.7), Spot number 13 (MW 33.1 pH 7.0), Spot number 14 (MW 31.6 pH 7.1), Spot number 17 (MW 33.1 pH 8.3) , M; protein marker
Fig. 3
Fig. 3
The Protein spots of Fasciola hepatica ES products, control sample (C9), in 2-DE gel. Spot number 1 (MW 53.2 pH 6.6), Spot number 5 (MW 31.6 pH 5.9), Spot number 8 (MW 31.6 pH 6.4), Spot number 9 (MW 30.0 pH 6.4), Spot number 11 (MW 33.1 pH 6.6), Spot number 12 (MW 33.6 pH 6.7), Spot number 13 (MW 33.1 pH 7.0), Spot number 14 (MW 31.6 pH 7.1), M; protein marker
Fig. 4
Fig. 4
The Protein spots of Fasciola hepatica ES products, test sample (T9), in 2-DE gel. Spot number 1 (MW 53.2 pH 6.6), Spot number 5 (MW 31.6 pH 5.9), Spot number 6 (MW 30.0 pH 5.9), Spot number 7 (MW 35 pH 6.3), Spot number 8 (MW 31.6 pH 6.4), Spot number 9 (MW 30.0 pH 6.4), Spot number 11 (MW 33.1 pH 6.6), Spot number 12 (MW 33.6 pH 6.7), Spot number 13 (MW 33.1 pH 7.0), Spot number 14 (MW 31.6 pH 7.1), Spot number 15 (MW 33.1 pH 7.6), Spot number 16 (MW 31.6 pH 7.6), Spot number 17 (MW 33.1 pH 8.3), M; protein marker
Fig. 5
Fig. 5
The Protein spots of Fasciola hepatica ES products, control sample (C10), in 2-DE gel. Spot number 1 (MW 53.2 pH 6.6), Spot number 5 (MW 31.6 pH 5.9), Spot num-ber 6 (MW 30.0 pH 5.9), Spot number 8 (MW 31.6 pH 6.4), Spot number 9 (MW 30.0 pH 6.4), Spot number 11 (MW 33.1 pH 6.6), Spot number 12 (MW 33.6 pH 6.7, Spot num-ber 18 (MW 32.0 pH 8.6), M; protein marker
Fig. 6
Fig. 6
The Protein spots of Fasciola hepatica ES products, test sample (T10), in 2-DE gel. Spot number 1 (MW 53.2 pH 6.6), Spot number 2 (MW 32 pH 5.2), Spot number 3(MW 30.0 pH 5.3), Spot number 4 (MW 35.0 pH 5.8), Spot number 5 (MW 31.6 pH 5.9), Spot number 6 (MW 30.0 pH 5.9), Spot number 7 (MW 35 pH 6.3), Spot num-ber 8 (MW 31.6 pH 6.4), Spot number 9 (MW 30.0 pH 6.4), Spot number 10 (MW 35.0 pH 6.5), Spot number 11 (MW 33.1 pH 6.6), Spot number 12 (MW 33.6 pH 6.7), Spot number 13 (MW 33.1 pH 7.0), Spot number 14 (MW 31.6 pH 7.1), Spot number 15 (MW 33.1 pH 7.6), Spot number 16 (MW 31.6 pH 7.6), Spot number 17 (MW 33.1 pH 8.3), Spot number 18 (MW 32.0 pH 8.6), M; protein marker
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