摘要
目的探讨胆盐输出泵(BSEP)在胆道梗阻-再通大鼠肝组织中的表达及意义。方法将54只Wistar大鼠随机分为假手术(Sham)组、胆总管结扎(CBDL)组、胆道再通(BR)组,测定各时相点血清丙氨酸氨基转移酶(ALT)、总胆汁酸(TBA)以及肿瘤坏死因子(TNF-α)的水平,采用RT-PCR方法和二步法免疫组化染色分别在基因转录、蛋白表达及肝细胞膜定位上检测BSEP在大鼠肝组织中的表达情况,运用Mias99病理图像分析系统半定量检测BSEP的平均积分光密度(IOD),与TBA和TNF-α作相关分析。结果CBDL术后大鼠血清ALT,TBA及TNF-α显著增高(P<0.05);BR术后上述指标逐步好转,7d时仍显著高于Sham组水平(P<0.05)。BSEP mRNA水平随着梗阻时间的延长明显降低(P<0.05),CBDL术后14d仅微弱表达,梗阻解除后缓慢增高;BR术后7d仍未恢复至Sham组水平。免疫组化染色显示BSEP表达定位于肝细胞膜上,变化规律与前述一致,并且与TBA和TNF-α呈显著负相关(r1=-0.9257,r2=-0.8536,P<0.05)。结论胆道梗阻-再通大鼠模型中胆盐的代谢与BSEP水平密切相关,而TNF-α可能是影响BSEP变化的重要因素。
Objective To explore the role of bile salt export pump （BSEP） in rat liver with bile duct obstruction and bile reflow. Methods Fifty-four male Wistar rats were randomizedly divided into three groups, sham group, common bile duct ligation （CBDL） group and bile reflow （BR） group to detect level of serum ALT, total bile acids （TBA） and tumor necrosis factor-α （TNF-α） at every time point. Expression of BSEP at level of gene transcription, protein expression and hepatic cell membrane localization was measured by using RT-PCR and immunohistochemistry method. Mean BSEP-IOD was detected semi-quantitatively by using Mias99 pathological image analysis system. Meanwhile, correlation of BSEP-IOD with serum TBA and TNF-α was performed. Results After CBDL, levels of serum ALT, TBA and TNF-α were ascended significantly （ P〈0.05）, which got a gradually favourable turn but was still higher than that in sham group seven days after bile reflow （ P 〈0.05）. The expression of BSEP mRNA decreased as bile duct obstruction elongated （ P〈0.05）, increased slowly 14 days after CBDL and posterior to obstruction relief but did not reach level of sham group seven days after BR. Immunohistochemistry results showed a similar tendency to above-mentioned change, ie, expression of BSEP was restrictively located on the hepatocyte membrane, which was negatively correlated with TBA and TNF-α （ r1 = -0. 9257, r2 =-0. 8536, P 〈0.05）. Conclusion BSEP may be responsible for bile salt metabolism in rat model of bile duct obstruction and bile reflow, and TNF-α plays an important role in regulating BSEP expression.
出处
《消化外科》
CSCD
2006年第6期451-454,共4页
Journal of Digestive Surgery
基金
全军医学科研"十五"计划面上项目(No.01MA176)
关键词
胆盐输出泵
肿瘤坏死因子-Α
胆道梗阻
再通
肝组织
大鼠
Bile salt export pump
Tumor necrosis factor a
Biliary tract obstruction
Recanalization
Liver cells
Rats
