Study on Glutathione S-Transferase (GST) Inhibition Assay by Triclabendazole. І: Protoscoleces (Hydatid Cyst; Echinococcus granulosus) and Sheep Liver Tissue
Hydatid disease is a term used to refer infection with the methacestode of Echinococcus granulosus parasite in humans, and echinococcusis is restricted to infection with the adult stage in carnivores.Glutathione S-Transferase (GST) represents the major class of detoxification enzymes from helminth p...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Tehran University of Medical Sciences
2005-06-01
|
| Series: | Iranian Journal of Public Health |
| Subjects: | |
| Online Access: | http://journals.tums.ac.ir/PdfMed.aspx?pdf_med=/upload_files/pdf/907.pdf&manuscript_id=907 |
| _version_ | 1818865980081700864 |
|---|---|
| author | MA Seyyedi A Farahnak M Jalali MB Rokni |
| author_facet | MA Seyyedi A Farahnak M Jalali MB Rokni |
| author_sort | MA Seyyedi |
| collection | DOAJ |
| description | Hydatid disease is a term used to refer infection with the methacestode of Echinococcus granulosus parasite in humans, and echinococcusis is restricted to infection with the adult stage in carnivores.Glutathione S-Transferase (GST) represents the major class of detoxification enzymes from helminth parasites such as Echinococcus protoscoleces (PSC) and it is candidate for chemotherapeutic and vaccine design. Therefore, GST of protoscoleces could be a target for evaluation of drug effect as triclabendazole in hydatid cyst. For this purpose, GST enzymes were purified from protoscoleces of hydatid cyst and sheep liver tissue by glutathione affinity chromatography using a wash-batch method and subsequently detected their SDS-PAGE pattern. Afterward, GST specific activity levels were assayed in the whole extract and purified solutions spectrophotometrically at 30°C with reduced glutathione (GSH) and 1-chloro-2, 4-dinitrobenzen (CDNB) substrate. Finally, GST inhibition assay was investigated in the solutions by powder and bolus of triclabendazole. GST fraction as a 26 kDa (MW) band was obtained on SDS-PAGE. The level of GST specific activity in purified solutions was detected 10.24 µmol/min/mg proteins for protoscoleces and 37.84 µmol/min/mg protein for liver tissue. Comparison of the effect of powder and bolus of triclabendazole in solutions revealed inhibition concentration (IC50) 8.71 and 11.16 µg/ml for protoscoleces GST and 8.65 and 9.70 µg/ml for liver tissue GSTs, respectively. These findings suggest the possibility of selective inhibition of protoscoleces. GSTs by triclabendazol in vitro and use of these results for understanding of its molecular effect in vivo. |
| first_indexed | 2024-12-19T10:56:10Z |
| format | Article |
| id | doaj.art-302cc89984ee496c94970b54456d4a64 |
| institution | Directory Open Access Journal |
| issn | 2251-6085 |
| language | English |
| last_indexed | 2024-12-19T10:56:10Z |
| publishDate | 2005-06-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Public Health |
| spelling | doaj.art-302cc89984ee496c94970b54456d4a642022-12-21T20:24:48ZengTehran University of Medical SciencesIranian Journal of Public Health2251-60852005-06-013413846Study on Glutathione S-Transferase (GST) Inhibition Assay by Triclabendazole. І: Protoscoleces (Hydatid Cyst; Echinococcus granulosus) and Sheep Liver TissueMA SeyyediA FarahnakM JalaliMB RokniHydatid disease is a term used to refer infection with the methacestode of Echinococcus granulosus parasite in humans, and echinococcusis is restricted to infection with the adult stage in carnivores.Glutathione S-Transferase (GST) represents the major class of detoxification enzymes from helminth parasites such as Echinococcus protoscoleces (PSC) and it is candidate for chemotherapeutic and vaccine design. Therefore, GST of protoscoleces could be a target for evaluation of drug effect as triclabendazole in hydatid cyst. For this purpose, GST enzymes were purified from protoscoleces of hydatid cyst and sheep liver tissue by glutathione affinity chromatography using a wash-batch method and subsequently detected their SDS-PAGE pattern. Afterward, GST specific activity levels were assayed in the whole extract and purified solutions spectrophotometrically at 30°C with reduced glutathione (GSH) and 1-chloro-2, 4-dinitrobenzen (CDNB) substrate. Finally, GST inhibition assay was investigated in the solutions by powder and bolus of triclabendazole. GST fraction as a 26 kDa (MW) band was obtained on SDS-PAGE. The level of GST specific activity in purified solutions was detected 10.24 µmol/min/mg proteins for protoscoleces and 37.84 µmol/min/mg protein for liver tissue. Comparison of the effect of powder and bolus of triclabendazole in solutions revealed inhibition concentration (IC50) 8.71 and 11.16 µg/ml for protoscoleces GST and 8.65 and 9.70 µg/ml for liver tissue GSTs, respectively. These findings suggest the possibility of selective inhibition of protoscoleces. GSTs by triclabendazol in vitro and use of these results for understanding of its molecular effect in vivo.http://journals.tums.ac.ir/PdfMed.aspx?pdf_med=/upload_files/pdf/907.pdf&manuscript_id=907Echinococcus granulosus protoscolecesTriclabendazole |
| spellingShingle | MA Seyyedi A Farahnak M Jalali MB Rokni Study on Glutathione S-Transferase (GST) Inhibition Assay by Triclabendazole. І: Protoscoleces (Hydatid Cyst; Echinococcus granulosus) and Sheep Liver Tissue Iranian Journal of Public Health Echinococcus granulosus protoscoleces Triclabendazole |
| title | Study on Glutathione S-Transferase (GST) Inhibition Assay by Triclabendazole. І: Protoscoleces (Hydatid Cyst; Echinococcus granulosus) and Sheep Liver Tissue |
| title_full | Study on Glutathione S-Transferase (GST) Inhibition Assay by Triclabendazole. І: Protoscoleces (Hydatid Cyst; Echinococcus granulosus) and Sheep Liver Tissue |
| title_fullStr | Study on Glutathione S-Transferase (GST) Inhibition Assay by Triclabendazole. І: Protoscoleces (Hydatid Cyst; Echinococcus granulosus) and Sheep Liver Tissue |
| title_full_unstemmed | Study on Glutathione S-Transferase (GST) Inhibition Assay by Triclabendazole. І: Protoscoleces (Hydatid Cyst; Echinococcus granulosus) and Sheep Liver Tissue |
| title_short | Study on Glutathione S-Transferase (GST) Inhibition Assay by Triclabendazole. І: Protoscoleces (Hydatid Cyst; Echinococcus granulosus) and Sheep Liver Tissue |
| title_sort | study on glutathione s transferase gst inhibition assay by triclabendazole і protoscoleces hydatid cyst echinococcus granulosus and sheep liver tissue |
| topic | Echinococcus granulosus protoscoleces Triclabendazole |
| url | http://journals.tums.ac.ir/PdfMed.aspx?pdf_med=/upload_files/pdf/907.pdf&manuscript_id=907 |
| work_keys_str_mv | AT maseyyedi studyonglutathionestransferasegstinhibitionassaybytriclabendazoleíprotoscoleceshydatidcystechinococcusgranulosusandsheeplivertissue AT afarahnak studyonglutathionestransferasegstinhibitionassaybytriclabendazoleíprotoscoleceshydatidcystechinococcusgranulosusandsheeplivertissue AT mjalali studyonglutathionestransferasegstinhibitionassaybytriclabendazoleíprotoscoleceshydatidcystechinococcusgranulosusandsheeplivertissue AT mbrokni studyonglutathionestransferasegstinhibitionassaybytriclabendazoleíprotoscoleceshydatidcystechinococcusgranulosusandsheeplivertissue |