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败血症休克大鼠肝脏内质网钙ATP酶磷酸化及去磷酸化的改变

高爱国, 董林旺, 王雪青, 刘乃奎, 唐朝枢, 刘茂生

(北京医科大学心血管基础研究所休克研究室，北京100083)；美国圣路易斯大学药理生理系)

摘要

该文观察子早期，晚期败血症休克大鼠肝脏内质网钙ATP酶磷酸化的改变。败血症休克模型由结扎二鼠直肠并穿孔(CLP)的方法复制。采用蔗糖密度梯度离心法分离肝脏内质网。由聚丙烯酞胺凝胶龟矫rgDS-PAGE )鉴定肝脏内质网钙ATP酶磷酸化中间产物。结果发现，肝脏粗面内质网，中间1x1质坷，滑面内质网钙ATP酶磷酸化能力在败血症休克早期降低15=23 0 o，晚期降低17-27'0 o (P<0. 05)。动力学分析发现，早期、晚期败血症休克时，钙离子、ATP对钙ATP酶磷酸化的最大反应速度都明显降低，而Km值无明显变化。SDS-PAGE分析证实，肝脏内质网钙ATP酶磷酸化的中间产物是一个分子量为115 kD的蛋白质。结论认为:败血症休克时，肝脏内质网钙ATP酶磷酸化能力明显减低，去磷酸化能力无明显变化，可能是败血症休克时肝细胞钙稳态失衡进而引起机体代谢改变的主要原因之一。

关键词： 败血症休克; 内质网; 磷酸化; 去磷酸化; 钙离子

IMPAIRMENT IN THE PHOSPHORYLATION OF Ca~(2+)一TRANSPORT ATPase OF RAT LIVER ENDOPLASMIC RETICULUM DURING SEPSIS

GAO AI-GUO, DONG LIN-WANG, WANG XUE-QING, LIU NAI-KUI , TANG CHAD-SHU, LIU MAW-SHUNG

Laboratory of Shock Research, Institute of Cardiovascular Research Beijing Medical University, Beijing 100083(Department of Physiology and Pharmacology, St. Louis University School of Medicine))

Abstract

The phosphorylation of Cat+-transport ATPase of rat liver endoplasmic reticulum (ER)during early and late septic shock induced by cecum ligation and puncture (CLP) was investigated by determining incorporation of [y-"P] ATP into Cat+-ATP phosphoprotein intermediate. Hepatic endoplasmic reticulum was isolated by differential centrifugation with sucrose density gradient. The Cal+-ATPase phosphoprotein intermediate was identified by SDS-PAGE. The results showed that the phosphorylation of Cat+-ATPase(115 kD) was decreased respectively by 15-23%(P<0. 05) and 17一27%(P<0. 05) at 9 h (early sepsis) and 18 h (late sepsis)，following the CLP in the rough，intermediate and smooth ER preparations. Kinetic analysis using rough ERshowed that the Vm$X for Ca~(2+) and for ATP for the phosphorylation of Cal+一ATPase were decreased dramatically during early and late sepsis，but without changes in the Km values. These results demonstrate that the phosphorylation of the phosphoprotein inter-
mediate of Ca~(2+)ATPase in rat liver was impaired during different phases of sepsis.

Key words: shock;septic;endoplasmic reticulum;phosphorylation;dephosphorylatipn

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引用本文：

高爱国, 董林旺, 王雪青, 刘乃奎, 唐朝枢, 刘茂生. 败血症休克大鼠肝脏内质网钙ATP酶磷酸化及去磷酸化的改变[J]. 生理学报 1996; 48 (3): .

GAO AI-GUO, DONG LIN-WANG, WANG XUE-QING, LIU NAI-KUI , TANG CHAD-SHU, LIU MAW-SHUNG. IMPAIRMENT IN THE PHOSPHORYLATION OF Ca~(2+)一TRANSPORT ATPase OF RAT LIVER ENDOPLASMIC RETICULUM DURING SEPSIS. Acta Physiol Sin 1996; 48 (3): (in Chinese with English abstract).