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谷胱甘肽S-转移酶(GSTs)作为肝包虫囊肿体外诊断生物标志物的活性测定

Activity Assay of Glutathione S-Transferase (GSTs) Enzyme as a Diagnostic Biomarker for Liver Hydatid Cyst in Vitro.

作者信息

Moatamedi Pour Lila, Farahnak Ali, Molaei Rad Mohamadbagher, Golmohamadi Taghi, Eshraghian Mohamadreza

机构信息

1. Dept. of Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Iran.

2. Dept. of Biochemistry, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.

出版信息

Iran J Public Health. 2014 Jul;43(7):994-9.

Abstract

BACKGROUND

The aim of this study was to detect the Glutathione S-Transferase(GST) enzyme activity of healthy / cystic liver as a diagnostic biomarker for hydatidosis. In order to compare with liver tissue, the level of the GSTs enzyme activity of parasite was also determined.

METHODS

Parasites were collected from sheep liver tissue with hydatid cysts at a local abattoir and washed with PBS buffer. Collected parasites and liver tissues were sonicated or homogenized respectively. Extract solution samples were centrifuged and stored at - 20°C. GST enzyme activities were measured in the extract of parasite and liver tissue samples (healthy and infected livers). Protein amounts and protein bands were detected using Bradford and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) methods respectively. To determine significant difference between two groups, two-sample t-test was performed.

RESULTS

GST specific activities of healthy / infected livers and parasites were estimated 304, 1297 and 146 U/ml/mg respectively. Significant higher GST specific activities in cystic liver than healthy liver was observed (P <0.05). T-test analysis showed GST activity of parasite was lower than healthy liver tissue. SDS-PAGE showed GST protein bands with 24 kDa in parasite samples and25 kDa in liver tissues.

CONCLUSION

GST activity incystic liver tissue could be concerned as a biomarker for hydatid cyst diagnosis with other hydatid disease parameters.

摘要

背景

本研究旨在检测健康/囊性肝脏的谷胱甘肽S-转移酶(GST)酶活性,作为包虫病的诊断生物标志物。为了与肝脏组织进行比较,还测定了寄生虫的GSTs酶活性水平。

方法

从当地屠宰场的患有包虫囊肿的绵羊肝脏组织中收集寄生虫,并用PBS缓冲液洗涤。分别对收集的寄生虫和肝脏组织进行超声处理或匀浆。提取液样品离心后保存在-20°C。在寄生虫和肝脏组织样品(健康肝脏和感染肝脏)的提取物中测量GST酶活性。分别使用Bradford法和十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)法检测蛋白质含量和蛋白条带。为了确定两组之间的显著差异,进行了双样本t检验。

结果

健康/感染肝脏和寄生虫的GST比活性分别估计为304、1297和146 U/ml/mg。观察到囊性肝脏中的GST比活性显著高于健康肝脏(P<0.05)。t检验分析表明寄生虫的GST活性低于健康肝脏组织。SDS-PAGE显示寄生虫样品中GST蛋白条带为24 kDa,肝脏组织中为25 kDa。

结论

囊性肝脏组织中的GST活性可与其他包虫病参数一起作为包虫囊肿诊断的生物标志物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97c8/4401064/3795fb1ca770/IJPH-43-994-g001.jpg

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