肝及肝癌组织缺血再灌注后枯否细胞的凋亡及意义

Apoptosis and Significance of Kupffer Cells in the Liver and Hepatocarcinoma Tissues after Ischemia and Reperfusion

目的研究肝癌组织缺血再灌注后一氧化氮(NO)和一氧化氮合成酶(iNOS)的改变、枯否细胞(Kupffer cell)的凋亡及其关系。方法通过超声引导将VX2肿瘤组织混悬液穿刺注射到新西兰兔肝脏左中叶,建立肝脏肿瘤模型,用无损伤血管钳阻断肿瘤所在肝叶的肝动脉分支60 min后去除血管阻断恢复血流,随机将模型动物分为缺血再灌注前(对照)、缺血再灌注后0 min1、h1、d3、d、1周6个时间组,取肝脏组织和肿瘤组织,分别测定NO和NOS的含量。用HE染色法观察肝组织和肝癌组织中的枯否细胞凋亡情况。结果肝组织再灌注后0min开始NO和iNOS浓度均明显下降,至再灌注7 d仍处于较低水平(44.41±2.0,3.70±0.2)。在肝癌组织,除NO浓度下降外,iNOS浓度从缺血再灌注0 min至7 d则明显升高(18.45±1.9)。肝组织和肝癌组织的枯否细胞凋亡显著增加,于再灌注1 d和1 h分别达最高峰(13.69±3.1,13.66±1.8)。结论枯否细胞对肝癌组织缺血再灌注过程中的iNOS及NO的产生、细胞损伤与凋亡起重要作用。

Objective To study the changes of nitric oxide （NO）, inducible nitric oxide synthases （iNOS） and apoptosis of Kupffer cells, and their relationship following ischemia and reperfusion in hepatocarcinoma and normal liver tissues. Methods The hepatocarcinoma animal models were established by ultrasonography-guided implantation of VX2 tumor tissue pieces into the left-middle lobe of liver of rabbits. The animals were subjected to 60 minutes clamp -induced ischemia of hepatic artery distributing in the left-middle lobe followed by the reperfusion at 0 min, 1 h, 1 d, 3 d and 7 d, respectively. Taking normal hepatic tissues and cancer tissues, the concentrations of NO and iNOS were determined and apoptotic changes were examined. Results The concentrations of NO and iNOS in normal liver tissues were low and kept until reperfusion for 7 days （44.41 ± 2.0,3.70 ± 0.2）. In hepatocarcinoma tissues, except of decrease of concentrations of NO, the concentrations of iNOS were significantly increased following reperfusion 0 min to 7 days（ 18.45 ± 1.9）. Apoptotic Kupffer cells in normal liver tissues and hepatocarcinoma tissues were increased and reached a maximum at 1 d and 1 h, respectively （ 13.69 ± 3.1,13.66 ± 1.8 ）. Conclusion Kupffer cells may play an important role in production of iNOS and NO, and injury of the hepatocarcinoma tissues following the ischemia and reperfusion.