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锌暴露对大鼠肝脏金属硫蛋白的诱导作用:剂量和时间依赖性研究。

Induction of Metallothionein in Rat Liver by Zinc Exposure: A Dose and Time Dependent Study.

作者信息

Garla Roobee, Kango Preeti, Gill Navneet Kaur, Garg M L

机构信息

Department of Biophysics, Panjab University, Chandigarh, 160014, India.

出版信息

Protein J. 2017 Oct;36(5):433-442. doi: 10.1007/s10930-017-9737-7.

Abstract

Metallothioneins (MTs) are low molecular weight ubiquitous metalloproteins with high cysteine (thiol) content. The intracellular concentration of zinc (Zn) is tightly regulated and MT plays a crucial role in it. The present study investigates the relationship between the Zn status (as a function of Zn concentration and time) in the rat liver and the occurrence of hepatic MT. For dose dependent study, four experimental groups, one control and three receiving different levels of metal supplementation, were chosen [Group 1 control and Group 2, Group 3, Group 4 receiving subcutaneous dose of 10, 50 and 100 mg of Zn/kg body weight (in the form of ZnSO·7HO), respectively]. For the time dependent expression of MT, again four experimental groups, i.e. Group 5 control and Group 6, Group 7, Group 8 receiving 50 mg of Zn/kg body weight (in the form of ZnSO·7HO) subcutaneously and sacrificed at different time intervals after last injection i.e. 6, 18, 48 h, respectively were chosen. Isolation of MT was done by using combination of gel filtration and ion exchange chromatography while characterization of MT fraction was carried in the wavelength range 200-400 nm. Expression of MT was studied by using Western blot analysis. The results revealed that the MT expression increases with increasing the dose of Zn administered and maximum at 18 h after last Zn injection. Accumulation of MT with increase dose would help in maintaining the intracellular Zn concentration by its sequestration which further reduces the possibility of undesirable binding of Zn to other proteins significantly and maintains Zn homeostasis. The maximum expression of MT at 18 h is indicative of its half life.

摘要

金属硫蛋白(MTs)是一类低分子量的、广泛存在的金属蛋白,富含半胱氨酸(硫醇)。细胞内锌(Zn)的浓度受到严格调控,MT在其中起着关键作用。本研究调查了大鼠肝脏中锌状态(作为锌浓度和时间的函数)与肝脏MT发生之间的关系。对于剂量依赖性研究,选择了四个实验组,一个对照组和三个接受不同水平金属补充的组[第1组为对照组,第2组、第3组、第4组分别皮下注射10、50和100 mg Zn/kg体重(以ZnSO₄·7H₂O的形式)]。对于MT的时间依赖性表达,同样选择了四个实验组,即第5组为对照组,第6组、第7组、第8组分别皮下注射50 mg Zn/kg体重(以ZnSO₄·7H₂O的形式),并在最后一次注射后的不同时间间隔(即分别为6、18、48小时)处死。通过凝胶过滤和离子交换色谱相结合的方法分离MT,同时在200 - 400 nm波长范围内对MT组分进行表征。通过蛋白质印迹分析研究MT的表达。结果显示,MT表达随着锌给药剂量的增加而增加,在最后一次注射锌后18小时达到最大值。MT随着剂量增加而积累,通过螯合作用有助于维持细胞内锌浓度,这进一步显著降低了锌与其他蛋白质发生不良结合的可能性,并维持锌的稳态。MT在18小时的最大表达表明了其半衰期。

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