内质网应激预处理对肝毒性药物导致原代大鼠肝细胞损伤的保护作用研究

Effect of Endoplasmic Reticulum Stress Preconditioning on Cytotoxicity of Clinically Relevant Hepatoxins in Primary Rat Hepatocytes

目的探讨大鼠肝细胞的内质网应激预处理对临床相关肝毒性药物肝细胞损伤的保护作用。方法 (1)将培养的原代大鼠肝细胞分为8组,分别予以不同浓度的衣霉素和毒胡萝卜素进行干预,尔后检测葡萄糖调节蛋白78 GRP78、葡萄糖调节蛋白94 GRP78。(2)衣霉素和毒胡萝卜素干预作用24 h后,可造成50%~75%的细胞死亡。对肝细胞进行预处理后,再使用顺铂、百草枯和利福平对细胞进行干预,检测细胞生存率。结果内质网应激诱导剂(毒胡萝卜素、衣霉素)对于GRP78和GRP94的表达表现出剂量相关的诱导作用,随着浓度的增加其表达量也逐渐增加,其变化具有明显的统计学意义(P<0.05)。而临床相关的毒性药物(顺铂、利福平、百草枯)对细胞的损伤可明显地因毒胡萝卜素和衣霉素的内质网应激预处理而减轻,其结果具有统计学意义。结论内质网应激预处理对临床相关肝毒性药物导致原代大鼠肝细胞损伤具有保护作用。这些结果提示在药物导致的肝损伤中内质网应激或许起到了一定作用。

Objective o study the protective effect of endoplasmic reticulum stress pretreatment on rat liver cells damaged by clinically-used hepatotoxic drugs. Methods （ 1 ） Cultured primary rat hepatocytes intervened with tunicamycin and thapsigargin at different concentrations were divided into eight groups. Glucose-regulated protein 78 （GRP78） and glucose-regulated protein 94（G^X）4） were detected. （2） 50% -75% liver cells were dead after 24h＇s intervention of tunicamycin and thapsigargin. The cell survival rate was detected after cisplatin, paraquat and rifampin were used. Re- sults The expression of GRP78 and GRP94, induced by endoplasmic reticulum stress revulsant carotene and tunicamy- cin, was increased with the concentration. The difference was of statistical significance （ P 〈 0.05）. The damage of tox- ic agents （cisplatin, rifampicin, paraquat）could be alleviated by endoplasmic reticulum stress pretreatment. Conclusion The endoplasmic reticulum stress pretreatment has a protective effect on primary rat hepatocyte injury caused by clinical- ly relevant liver toxicity of drugs.