In vitro assay of alkaline phosphatase enzyme activity in liver and fasciola hepatica parasite
Archives of Advances in Biosciences,
Vol. 8 No. 3 (2017),
13 June 2017
,
Page 13-17
https://doi.org/10.22037/jps.v8i3.11560
Abstract
The present work was designed to determine alkaline phosphatase (ALP) activity level in Fasciola hepatica parasite and Fasciola infected livers to evaluate the effects of liver infection on enzyme activity and to compare enzyme activity in liver and parasite. The sheep livers were collected and adult Fasciola hepatica parasites were isolated and washed with PBS buffer. Collected healthy and infected livers and parasite were homogenized and extract solutions were centrifuged and stored at -20°C. ALP enzymes activity was measured in the extract solution of samples. Proteins of the samples were measured and protein bands were detected through using Bradford and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) methods respectively. Independent two samples t-test was performed to determine significant difference between different groups. The mean values of the assayed ALP specific activities of infected and healthy livers and Fasciola hepatica parasite were estimated 0.163,0.133,0.048 U/mg protein/ml correspondingly. Gel electrophoresis (SDS-PAGE) of Fasciola hepatica and liver extract solution samples displayed different protein bands, including ALP enzyme. Statistical analysis did not show significant difference between enzyme’s activity in infected and healthy livers (P>0.05). However, the liver demonstrated more than ALP activity level to parasite (P<0.05). The findings show that ALP enzymes activity in Fasciola hepatica infected livers could not be concerned as a specific pathological biomarker in fascioliasis, but meanwhile this enzyme displays interest activity in parasite.
- Alkaline phosphatase
- Fasciola hepatica
- SDS-PAGE
- Protein
- Liver
How to Cite
References
Mas-Coma MS, Esteban JG, and Bargues MD. Epidemiology of human fascioliasis: a review and proposed new classification. Bull World Health Organ. 1999; 77(4): 340-46.
Farahnak A, Golmohamadi T, Eshraghian MR. In vitro Effects of Triclabendazole (TCBZ) on the Excretory-Secretory Products (ESP) of Fasciola spp Parasites. Acta Medica Iranica. 2012; 50(3): 164-168.
Farahnak, A., A. Golestani, and M. Eshraghian. Activity of Superoxide Dismutase (SOD) Enzyme in the Excretory-Secretory Products of Fasciola hepatica and F. gigantica Parasites. Iranian journal of parasitology. 2013; 8(1): 167-170.
Shrifi Y, Farahnak A, Golestani A, Eshraghian MR, Faridi A, Molaei rad MB. Triclabendazole effect on protease enzyme activity in the excretory-secretory products of Fasciola hepatica in vitro. Iran J Parasitol. 2014; 9(1): 107-113.
Swargiary A, Roy B, Ronghang B. Partial characterisation of alkaline phosphatase in Fasciolopsis buski–an intestinal fluke treated with crude extract of Alpinia nigra (Zingiberaceae). J Pharm Technol Drug Res. 2013; 2(5): 1-7.
Yan L, Shenghui Y, Jianhua X, Liang Y, Li C, Ju Z, Kegeng W, Sijie T, Zhengyang Y, Qingren Z. M13 phage peptide ZL4 exerts its targeted binding effect on schistosoma japonicum via alkaline phosphatase. Int J Clin Exp Pathol. 2015; 8(2) :1247-1258.
Shamban L, Patel B, Williams M. Significantly Elevated Liver Alkaline Phosphatase in Congestive Heart Failure. Gastroenterology Res. 2014; 7(2): 64-78.
Maizels R M, Blaxter ML, Robertson BD, Selkirk ME. Parasite antigens, parasite genes: a laboratory manual for molecular parasitology.London:CUP Archive;1991.
Murray RK, Granner DK, Mayer PA, Rodwell VW. Harper's Biochemistry. Norwalk: Appleton & Lange;1988.
Anderson PH, Berrett S, Brush PJ, Hebert CN, Parfitt JW, Patterson DS. Biochemical indicators of liver injury in calves with experimental fascioliasis. Vet Rec. 1977; 100(3): 43-45.
American Association for Clinical Chemistry. Alkaline Phosphatase (ALP)[Internet]. 2017 [Cited 22 March 2017]. Available from: https://labtestsonline.org/understanding/analytes/alp/tab/test/
Roe S. Protein purification techniques: a practical approach. Oxford: University Press Oxford; 2001.
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