缺血后处理减轻肝脏缺血再灌注过程中胆管细胞凋亡机制的研究  

Postconditioning attenuates bile duct cell damage by apoptosis from liver ischemia-reperfusion injury in rats

作　　者：郭怀斌[1] 刘小帅[1] 暴雷[1] 王春城[1] 王兰辉[1] 张万星[1] 

机构地区：[1]河北省人民医院普外三科,石家庄050051

出　　处：《中华实验外科杂志》2017年第7期1133-1135,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察胆管组织内B细胞淋巴瘤/白血病-2（bcl-2）及B细胞淋巴瘤/白血病-2相关X蛋白（bax）蛋白表达，探讨缺血后处理诱导肝脏缺血再灌注损伤过程中胆管细胞凋亡的机制。方法健康雄性SD大鼠24只，随机分为假手术（SO）组、缺血再灌注（IR）组、缺血后处理（IPO）组，每组8只。缺血后处理的方法：再灌注之前，预再灌注10 s、停灌注10 s，反复6次。检测血清中γ-谷氨酰胺转酞酶（γ-GT）活性，免疫组织化学法测定胆管组织中bcl-2和bax蛋白表达。用原位缺口末端标记法（TUNEL）检测胆管细胞凋亡指数。结果与IR组（34.64±3.28） U/L比较，IPO组血清内γ-GT活性（26.30±3.32） U/L降低，差异有统计学意义（P=0.001）；与IR组[（42.67±3.73）%]比较，IPO组胆管组织bcl-2蛋白[（77.94±5.25）%]表达增高，bax蛋白表达[（70.10±4.27）%比（90.17±2.12）%]减少，差异均有统计学意义（P=0.000）。IPO组凋亡指数（AI）[（58.90±2.75）%]较IR组[（79.31±1.12）%]下降（P=0.000），胆管细胞病理形态学损伤减轻。结论缺血后处理通过促进bcl-2蛋白表达、抑制bax蛋白表达，减少SD大鼠肝脏缺血再灌注过程中胆管细胞凋亡，减轻肝脏缺血再灌注对胆管细胞的损伤。Objective To study the protective effect of postconditioning on bile duct cell damage in the form of apoptosis during liver ischemia-reperfusion （IR） injury by observing the expression of B cell lymphoma/leukemia-2 （bcl-2） and B cell lymphoma/leukemia-2 associated X protein （bax） on bile ducts.Methods Thirty healthy male SD rats were randomly divided into three groups： sham-operation （SO） group, IR group and ischemic postconditioning （IPO） group. IPO was achieved by 6 brief reperfused/ischemia cycles before the persistent reperfusion procedure. The samples of blood and hepatic tissue of all groups were taken after experiment. The activity of γ-glutamyl transpeptidase （GT） in serum was determined. The expression of bcl-2 and bax on bile ducts was detected by immunohistochemistry. The apoptosis index on bile ducts was observed by TdT-mediated dUTP nick end labeling （TUNEL） methods.Results The activity of γ-GT was lower in IPO group [（26.30±3.32） U/L] than that in IR group [（34.64±3.28） U/L] with the difference being statistically significant （P=0.001）. As compared with the IR group, the expression of bcl-2 on bile ducts was up-regulated [（77.94±5.25）% vs. （42.67±3.73）%], and that of bax down-regulated [（70.10±4.27）% vs. （90.17±2.12）%] in IPO group, with the difference being statistically significant （P=0.000）. As compared with IR group, the apoptosis index in IPO group was decreased [（58.90±2.75）% vs. （79.31±1.12）%, P=0.000], and the pathologically morphologic damages of the bile duct cells were attenuated.Conclusion In the process of liver IR, the postconditioning can promote the expression of bcl-2 and inhibit the expression of bax on bile duct, which may withstand the happening of the bile duct cell apoptosis, thus attenuating the bile duct damage caused by liver IR injury.

关 键 词：再灌注损伤 缺血后处理 胆管 脱噬作用 B细胞淋巴瘤/白血病-2 B细胞淋巴瘤/白血病-2相关X蛋白 

分 类 号：R657.3[医药卫生—外科学]