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成年大鼠肝实质细胞和非实质细胞中金属硫蛋白的鉴定。

Identification of metallothionein in parenchymal and non-parenchymal liver cells of the adult rat.

作者信息

Sciortino C V, Failla M L, Bullis D B

出版信息

Biochem J. 1982 May 15;204(2):509-14. doi: 10.1042/bj2040509.

Abstract

Parenchymal and non-parenchymal cells were isolated from the livers of control, starved, Zn2+-injected and Cd2+-injected rats. Parenchymal cells were prepared by differential centrifugation after perfusion of the liver with collagenase. Non-parenchymal cells were separated from parenchymal cells by unit-gravity sedimentation and differential centrifugation. Yields of 2 x 10(8) non-parenchymal cells with greater than 95% viability and less than 0.2% contamination with parenchymal cells were obtained without exposing cells to Pronase. Metallothioneins-I and -II were identified in parenchymal cells and non-parenchymal cells from Zn2+-treated rats. The metallothionein contents of parenchymal cells, non-parenchymal cells and intact liver were quantified by a competitive 203Hg-binding assay. Administration of heavy-metal salts significantly increased the metallothionein content of both cell populations, although the concentration of the protein was approx. 2.5-fold greater in parenchymal cells than in non-parenchymal cells. Overnight starvation increased the metallothionein content of parenchymal cells without altering that of non-parenchymal cells. The potential significance of this differential response by different liver cell types with regard to the influence of Zn2+ on stress-mediated alterations in hepatic metabolism is discussed.

摘要

从对照、饥饿、注射Zn2+和注射Cd2+的大鼠肝脏中分离实质细胞和非实质细胞。在用胶原酶灌注肝脏后,通过差速离心制备实质细胞。通过单位重力沉降和差速离心将非实质细胞与实质细胞分离。在不将细胞暴露于链霉蛋白酶的情况下,获得了2×10(8)个非实质细胞,其活力大于95%,实质细胞污染率小于0.2%。在Zn2+处理的大鼠的实质细胞和非实质细胞中鉴定出金属硫蛋白-I和-II。通过竞争性203Hg结合试验对实质细胞、非实质细胞和完整肝脏中的金属硫蛋白含量进行定量。重金属盐的给药显著增加了两个细胞群体的金属硫蛋白含量,尽管蛋白质浓度在实质细胞中比在非实质细胞中大约高2.5倍。过夜饥饿增加了实质细胞的金属硫蛋白含量,而未改变非实质细胞的金属硫蛋白含量。讨论了不同肝细胞类型的这种差异反应对于Zn2+对肝脏代谢中应激介导的改变的影响的潜在意义。

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