title: Triclabendazole Effect on Protease Enzyme Activity in the Excretory- Secretory Products of Fasciola hepatica in Vitro.
abstract: BACKGROUND
  Fasciola hepatica is one of the most important helminthes parasites and triclabendazole (TCBZ) is routinely used for treatment of infected people and animals. Secreted protease enzymes by the F. hepatica plays a critical role in the invasion, migration, nutrition and the survival of parasite and are key targets for novel drugs and vaccines. The aim of study was to determine the protease activity of excretory- secretory products (ESP) of F. hepatica in the presence of TCBZ anthelmintic.
  
  
  METHODS
  F. hepatica helminthes were collected and cultured within RPMI 1640 [TCBZ treated (test) and untreated (control)] for 6 h at 37 °C. ESP of treated and control were collected, centrifuged and supernatants were stored at -20°C. Protein concentrations were measured according to Bradford method. Protease enzymes activities of ESP samples were estimated by using sigma's non-specific protease activity assay. ESP protein bands were detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).
  
  
  RESULTS
  Mean protein concentrations in control and treated of ESP samples were determined 196.1 ±14.52 and 376.4 ±28.20 μg/ml, respectively. Mean protease enzymes activities in control and treated were 0.37 ±0.1 and 0.089 ±0.03 U/ml, respectively. Significant difference between proteins concentrations and protease enzymes activities of two groups was observed (P<0.05). SDS-PAGE showed different patterns of protein bands between treated and control samples.
  
  
  CONCLUSIONS
  The TCBZ reduced secreted protease enzymes activities and possibly effects on invasion, migration, nutrition and particularly survival of the parasite in the host tissues.
authors: Department of Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors: Yosef
authors: Y
authors: Shrifi
authors: Department of Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors: Ali
authors: A
authors: Farahnak
authors: Department of Biochemistry, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors: Abolfazle
authors: A
authors: Golestani
authors: Department of Epidemiology and Biostatistics, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors: Mohamad Reza
authors: MR
authors: Eshraghian
authors: Department of Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors: Ashkan
authors: A
authors: Faridi
authors: Department of Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors: Mohamad Bagher
authors: MB
authors: Molaei Rad
keywords: Excretory-Secretory product; F. hepatica; Protease; Triclabendazole
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