摘要
目的探讨在阻塞性黄疸肝损害中 Bc(?)-2,Bax 基因的表达.方法结扎 Wistar 大鼠胆总管建立阻塞性黄疸大鼠动物模型.分为结扎后3,7,14,21 d 及阴性对照各组,每组8只.结扎后3,7,14,21 d 处死大鼠,门静脉抽血.离心后取上层血清行胆红素测定.HE 染色,光镜下观察阻塞性黄疸大鼠肝脏组织的病理改变.用末端脱氧核苷酸转移酶(TdT)介导的 dUTP 缺口末端标记技术(TUNEL)和免疫组织化学方法检测阻塞性黄疸大鼠肝脏组织细胞凋亡状态及凋亡相关基因 Bc(?)-2,Bax 蛋白的表达.结果胆总管结扎后,随结扎时间的延长胆红素值逐渐增高.光镜下,胆总管结扎3d 即见肝内胆管扩张,结扎7 d,肝细胞内或细胞间瘀胆,纤维组织增生;结扎14 d,纤维组织增生向肝小叶内扩展;结扎21d,纤维组织带增宽,肝脏结构广泛改变.TUNEL 检测表明,正常大鼠肝组织偶见细胞凋亡,结扎3 d 即见细胞凋亡增加,结扎7 d 明显增加,结扎14 d 后细胞凋亡达高峰,凋亡指数(AI)达58.2%±1.6%,各组 AI 有显著性差异(P<0.05),21 d 细胞凋亡数目明显减少.SABC 免疫组化检测,Bc(?)-2蛋白的表达,对照组、胆总管结扎3 d,7 d 均为阴性表达,14 d 弱阳性表达,21 d 强阳性表达;结扎14 d,Bc(?)-2蛋白弱阳性表达,AI 达高峰,结扎21 d Bc(?)-2强阳性表达,AI 下降.Bax蛋白的表达,对照组、胆总管结扎3 d 均可见表达,7 d 表达增多,14 d 明显增多;21 d 表达减少;结扎7 d,Bax 蛋白中阳性表达,AI 增高;结扎14 d,Bax 蛋白强阳性表达,AI 达高峰.表明在阻塞性黄疸过程中,Bc(?)-2蛋白表达越强,凋亡指数越少;Bax蛋白表达越强,凋亡指数越强.结论 Bc(?)-2和 Bax 蛋白均参与了阻塞性黄疸肝组织中细胞凋亡的调节,并在阻塞性黄疸肝损害的发生和发展中起重要作用.
AIM To explore the regulating mechanism of hepatic injury in obstructive jaundice (OJ). METHODS Forty adult male Wistar rats (190g-230g) were used for all experiments.All rats underwent laparotomy.Experimental obstructive jaundice (BDL) was induced by double ligation of the bile duct by a proximal ligature around the bile duct in the hilus of the liver and by a distal ligature close to its entry into the duodenum.The animals were randomly divided into two groups:those that underwent bile duct manipulation (sham operated) and those that underwent BDL for 3,7,14 and 21 days. We detected the bilirubin of blood and pathological changes of liver in alle rats.Then we observed the apoptotic status and the expression of apoptotic related gene Bcl-2,Bax protein in liver of alle rats by the terminal-deoxynucleotidyl mediated nick end labeling (TUNEL) and immunohistochemistry method. RESULTS The apoptotic rate of liver was related to times of obetructive jaundice.Apoptosis peaked from 14-day blle duct ligation,the apoptotic index (Al) being 58.2%± 1.6% (P<0.05).The stronger Bcl-2 expression,the weaker Bax expression and the less number of apoptotic cells in obstructive jaundice.The increase of bilirubin and pathological change of liver in obstructive jaundice were obviously. CONCLUSION Bcl-2 and Bax take part in the regulation and the occurrence and progression of hepatic injury in obstructive jaundice.
出处
《世界华人消化杂志》
CAS
2001年第8期911-914,共4页
World Chinese Journal of Digestology
