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目的探讨肝细胞毛细胆管面胆红素主要转运子多药耐药相关蛋白2(Mrp2)在胆道再通过程中的表达变化机制。方法对96只胆道梗阻及再通大鼠动物模型的286份肝组织样本进行放免检查TNF-α;进行免疫荧光和RT-PCR,在肝细胞毛细胆管面上染色定位Mrp2,计算机图像分析测定,半定量比较肝细胞毛细胆管面上Mrp2表达情况,并与肝组织匀浆TNF-α、血清胆红素作相关分析。结果Mrp2转运蛋白特异性定位于肝细胞膜的毛细胆管面,荧光染色呈线条形勾勒出毛细胆管轮廓,CBDL术后1、3、7和14 d其Mrp2表达(相对灰度)逐渐下降,由223.0±14.2降为211.0±13.7、167.0±8.7、112.0±11.2、78.0±7.2;RBF术后1、37、和14 d其Mrp2表达缓慢恢复,分别为102.0±9.7、143.0±8.1、177.0±12.4和212.0±15.4。并且随着肝组织匀浆TNF-α、血清胆红素的升高或降低而发生变化。结论胆道再通后肝细胞代谢胆红素的恢复与Mrp2转运蛋白的变化密切相关,且TNF-α可能是影响Mrp2变化的主要因素。
Objective To investigate the mechanism of expression of Mrp2, a major transporter of bilirubin in hepatocytes, during bile duct recanalization. Methods Totally 286 liver samples of 96 biliary obstruction and recanalization animal models were subjected to radioimmunoassay and RT-PCR. The mRNA of Mrp2 was detected by computer image analysis, Semi-quantitative comparison of the expression of Mrp2 on the surface of hepatocyte capillary bile duct, and liver tissue homogenate TNF-α, serum bilirubin for the correlation analysis. Results The Mrp2 transporter was localized on the surface of the capillary bile duct of liver cell membrane. The outline of capillary bile duct was outlined by fluorescent staining. The Mrp2 expression (relative gray level) gradually decreased from 223.0 ± 14.2 to 211.0 ± 13.7,167.0 ± 8.7,112.0 ± 11.2,78.0 ± 7.2; Mrp2 expression recovered slowly at 1, 37, and 14 days after RBF, which were 102.0 ± 9.7, 143.0 ± 8.1, 177.0 ± 12.4 and 212.0 ± 15.4. And with the liver homogenate TNF-α, serum bilirubin increased or decreased and changed. Conclusion The bilirubin recanalization is closely related to the change of Mrp2 transporter in the recovery of hepatocyte metabolism bilirubin, and TNF-α may be the main factor affecting Mrp2 change.