In vitro Effects of Triclabendazole (TCBZ) on the Excretory-Secretory Products (ESP) of Fasciola spp Parasites
Fascioliasis is an endemic disease in Iran and triclabendazole (TCBZ) is using for treatment of domestic animals and infected people. Excretory-secretory products (ESP) play an important role in the host biochemical defense by means of activities of detoxifying and antioxidant glutathione S-transferase (GST) and superoxide dismutase (SOD) enzymes respectively. Therefore, the aim of this comparative study was to evaluate fasciola protection against TCBZ drug by detection of enzymatic activities, GST and SOD, in TCBZ treated Fasciola hepatica / Fasciola gigantica and control ESP samples. F. gigantic and F. hepatica helminthes were collected and cultured within buffer media (TCBZ treated and untreated or control) for 4 h at 37 °C. Three TCBZ treated and 1 control ESP samples for each species were collected, centrifuged and supernatants were stored at -20°C. ESP samples protein concentrations were measured by Bradford method. SOD and GST enzymes activities of ESP samples were estimated photometrically. To determine the statistically significant difference between ESP of treated and control samples, t-test was conducted. ESP protein bands were detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Protein concentrations in treated F. hepatica and F. gigantica ESP samples were estimated 204.88, 428, 130.4 and 288.2, 488.2, 308.2 µg/ml respectively. Protein concentrations in control samples were estimated 488.18 and 124.8 ug/ml respectively. SOD enzyme specific activities level in treated F.hepatica and F. gigantica ESP samples were determined 0.14, 0.31, 3.96 and 11.11, 13.54, 19.95 U/mg/protein respectively. SOD activities level in control samples were detected 70.69 and 10.92 U/mg/protein. GST specific activities level in treated F.hepatica and F. gigantica ESP samples were calculated 25.3, 85.5, 37.3 and 1823, 1314.3, 1320.8 U/mg respectively. GST activities levels in control samples were detected 98.6 and 1083.9 U/mg/protein respectively. Statistical analysis reveal the significant different between proteins concentrations, GST and SOD enzyme specific activities of TCBZ treated ESP samples of F. gigantic in comparison to the control samples (P0.05). There is no difference between SDS-PAGE results of treated and control samples. Based on the results of the present work, significant increase of enzymatic activities of GST and SOD in TCBZ treated F. gigantica ESP, it seems, the protection of this species against drug is higher than F. hepatica.
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