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片形吸虫属寄生虫和肝组织提取物中超氧化物歧化酶(SOD)活性测定

Superoxide Dismutase (SOD) Enzyme Activity Assay in Fasciola spp. Parasites and Liver Tissue Extract.

作者信息

Assady M, Farahnak A, Golestani A, Esharghian Mr

机构信息

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

出版信息

Iran J Parasitol. 2011 Dec;6(4):17-22.

Abstract

BACKGROUND

The purpose of this comparative study was to detect superoxide dismutase (SOD) activities in Fasciola hepatica, F. gigantica parasites, infected and healthy liver tissues in order to determine of species effects and liver infection on SODs activity level.

METHODS

Fasciola spp. parasites and sheep liver tissues (healthy and infected liver tissues), 10 samples for each, were collected, homogenized and investigated for protein measurement, protein detection and SOD enzyme activity assay. Protein concentration was measured by Bradford method and SODs band protein was detected on SDS-PAGE. SODs activity was determined by iodonitrotetrazolium chloride, INT, and xanthine substrates. Independent samples t-test was conducted for analysis of SODs activities difference.

RESULTS

Protein concentration means were detected for F. hepatica 1.3 mg/ ml, F. gigantica 2.9 mg/ml, healthy liver tissue 5.5 mg/ml and infected liver tissue 1.6 mg/ml (with similar weight sample mass). Specific enzyme activities in the samples were obtained 0.58, 0.57, 0.51, 1.43 U/mg for F. hepatica, F. gigantica, healthy liver and infected liver respectively. Gel electrophoresis of Fasciola spp. and sheep liver tissue extracts revealed a band protein with MW of 60 kDa. The statistical analysis revealed significant difference between SOD activities of Fasciola species and also between SOD activity of liver tissues (P<.05).

CONCLUSION

Fasciola species and liver infection are effective causes on SOD enzyme activity level.

摘要

背景

本比较研究的目的是检测肝片吸虫、巨片吸虫寄生虫、受感染和健康肝脏组织中的超氧化物歧化酶(SOD)活性,以确定物种效应和肝脏感染对SOD活性水平的影响。

方法

收集肝片吸虫属寄生虫和绵羊肝脏组织(健康和受感染肝脏组织)各10个样本,进行匀浆,并对蛋白质含量测定、蛋白质检测和SOD酶活性测定进行研究。采用Bradford法测定蛋白质浓度,在SDS-PAGE上检测SOD条带蛋白。通过碘硝基四氮唑蓝(INT)和黄嘌呤底物测定SOD活性。采用独立样本t检验分析SOD活性差异。

结果

检测到肝片吸虫的蛋白质浓度平均值为1.3mg/ml,巨片吸虫为2.9mg/ml,健康肝脏组织为5.5mg/ml,受感染肝脏组织为1.6mg/ml(样本重量相似)。样本中的比酶活性分别为肝片吸虫0.58U/mg、巨片吸虫0.57U/mg、健康肝脏0.51U/mg、受感染肝脏1.43U/mg。肝片吸虫属和绵羊肝脏组织提取物的凝胶电泳显示一条分子量为60kDa的条带蛋白。统计分析显示,肝片吸虫物种的SOD活性之间以及肝脏组织的SOD活性之间存在显著差异(P<0.05)。

结论

肝片吸虫物种和肝脏感染是影响SOD酶活性水平的有效因素。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1eef/3279904/4200f3ec59ef/IJP-6-017-g001.jpg

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