精密肝切片技术在谷胱甘肽抗氯化镉肝毒性研究中的应用

为了解精密肝切片技术应用于毒物及药物代谢体外研究的可行性 ,利用精密肝切片技术 (PCLS) ,观察毒物氯化镉 (CdCl2 )的肝毒作用及谷胱甘肽 (GSH)的拮抗效应。于最适切片与培养条件下 ,观察CdCl20 1、0 5、2 5mmol L对肝切片活力、NO分泌的影响 ,及以CYP2E1、CYP3A4为代表的I相酶和UDPGT、GST为代表的II相酶活性的变化。同时观察GSH 2 0 ,4 0 ,8 0mmol L对CdCl2 肝毒作用的逆转效应。结果显示 ,CdCl2显著降低肝切片活力 ;在 2 5mmol L时 ,CYP含量和CYP3A4活性分别较对照组降低了 4 7 6 %、6 6 0 % ,UDPGT 1、UDPGT 2和GST分别比对照组下降了 5 0 0 %、35 6 %、2 2 6 % ;而CYP2E1活性比对照组升高了75 8%。GSH 2 0mmol L时 ,CYP含量、CYP2E1、CYP3A4和UDPGT 2活性等指标接近正常水平。上述结果重复 2～ 3次变化趋势一致。提示精密肝切片培养系统用于CdCl2 肝毒作用和GSH干预效应体外研究时 ,具有稳定性和敏感性的特点。表明PCLS是一种好的肝脏药理毒理学研究的体外模型系统

Application of precision-cut liver slice to the study of the effect of glutathione on cadmium chloride hepatotoxicity

In order to evaluate the feasibility of application of precision cut liver slice(PCLS) to the study of the effect of glutathione on cadmium chloride hepatotoxicity, slices were cultured with CdCl 2 and GSH. Slices activity and drug metabolic function was estimated by assay of slice viability, NO secretion, and phase I and phase II enzyme activity. Results showed that the slices activity was significantly inhibited by CdCl 2. Various phase I and phase II enzymes activity of slices were obviously reduced by CdCl 2, but aniline hydroxidase activity was enhanced. GSH could distinctly reversed the change of slices induced by CdCl 2. Therefore, it could be concluded that PCLS culture system was sensitive and steady, and was a good model for pharmacological and toxicological investigation of liver in vitro.