创伤后肝细胞凋亡与坏死的检测及其意义

Detection of posttrauma apoptosis and necrosis of hepatocytes and its significance

目的 探讨严重创伤后肝细胞凋亡及坏死在急性肝功能障碍发病机制中的作用。方法复制多发性骨折合并休克的大鼠创伤模型,采用 Annexin－V－Flous、碘化丙锭(propidium iodid,PI)双标法经流式细胞仪检测创伤后各时间点肝细胞凋亡与坏死的数量变化,结合光镜、电镜和电泳观察细胞凋亡与坏死,并与肝功能变化相比较。结果创伤后早期肝细胞即发生凋亡和坏死,坏死肝细胞的数量进行性升高,与肝功能变化显著呈正相关;凋亡肝细胞在创伤后3h达高峰,部分凋亡肝细胞发生继发性坏死,其数量与肝功能变化显著正相关。结论肝细胞坏死与凋亡是严重创伤后肝功能损害的重要原因,坏死肝细胞是肝功能损害的直接因素,凋亡肝细胞通过发生继发性坏死加重肝功能损害。

Objective To study the role of apoptosis and necrosis of hepatocytes in the pathogenis of acute liver dysfunctionfollowing severe trauma in rats. Methods Rat models of multiple fractures complicated by shock were established, in whichapoptotic and necrotic hepatocytes were detected by flow cytometry (FCM) via double-staining technique of Annexin-V-flousand propidium iodide (PI). Observation of the pathological changes in the hepatocytes was also conducted by means of lightand electron microscopy and electronphoresis respectively, and the correlation of these changes with liver function were eval-uated. Results Hepatocytes underwent both apoptosis and necrosis in rat models of severe theuma. The quantities ofnecrotichepatocytes increased progressively in positive correlation with the degree ofhepatic injury, reaching the peak at 3 h posttrau-ma. A proportion of the apoptotic cells underwent secondary necrosis, the number of the necrotic cells positively correlatedwith liver dysfunction deterioration. Conclusions Both necrosis and apoptosis are important factors responsible for liver func-tion damages following severe trauma. Necrosis of hepatic cells directly causes liver function damages that are further aggra-vated by secondary necrosis among apoptotic hepatocytes.