人肝刺激因子对CCl_4损伤大鼠离体肝细胞内钙和钾离子稳态的影响

EFFECT OF HUMAN HEPATIC STIMULATOR SUBSTANCE ON INTRACELLULAR CALCIUM AND POTASSIUM HOMEOSTASIS IN CCl_4-INTOXICATION DISSOCIATED HEPATOCYTES

本工作从自愿流产孕妇的胎儿取肝,按照LaBrecque法提取人肝刺激因子(human hepaticstimulator substance,hHSS)。用荧光探针Fura-2/AM测定离体肝细胞内游离钙,用离子分析仪测细胞染毒(四氯化碳CCl_4)前后基质中钾离子含量,观察hHSS对染毒肝细胞内Ca^(2+)和K^+稳态的影响,并测定肝细胞存活率和细胞内转氨酶(ALT)的漏出作为佐证。结果表明,人胎肝中含有hHSS,hHSS能提高离体肝细胞的存活率,维持肝细胞内游离钙的相对恒定,减少细胞内钾离子和ALT的漏出。这些结果提示,hHSS可保护肝细胞内钙,钾离子稳态和肝细胞膜的稳定,从而加强大鼠离体肝细胞抗CCl_4的损伤。

The human hepatic stimulator substance (hHSS) was extracted from abortive fetalliver according to the method of LaBrecque. The intracellular free calciumion was mea-sured by fluorescent probes Fura--2/AM. Rat hepatocytes were isolated from liver bythe method of Seglen and intoxicated by CCl_4 vapour, then the changes of [Ca^(2+)]_i, theleakage of intracellular K^+, ALT and viability of hepatocytes were observed. The re-sults were as follows: The hHSS does exist in the human fetal liver, hHSS could increasethe viability of intoxiacated hepatocytes, maintain the intracellular calcium homeosta-sis, and decrease the leakage of intracellular potassium and ALT into the culture medium. These results indicate that hHSS could protect hepatocytes against CCl_4 throughmaintaining calcium homeostasis, preventing potassium leakage and sustaining stabilityof hepatocyte polasmalemma.