胆道梗阻肝损伤早期NF-κB活性变化的影响及意义

Effect of cholestasis injury on nuclear factor-κB activation during early stage following obstruction of bile duct

目的探讨胆道梗阻导致的胆汁淤积性肝损伤时NF-κB活性变化的意义。方法将20只大鼠随机分为假手术组(SO组)和梗黄试验组(Ja组),Ja组通过胆总管结扎建立大鼠胆道梗阻模型。所有大鼠均于术后7 d处死。采集血清测定血清谷丙转氨酶(ALT)、总胆红素(TBIL)和总胆汁酸(TBA)水平。免疫组化染色测定肝组织NF-κB p65含量。RT-PCR测定肝脏组织中TNF-αmRNA水平。结果 Ja组ALT水平明显高于SO组(145.7±33.5μmol/L vs 40.7±7.3μmol/L,P<0.001),Ja组的TBIL和TBA水平较SO组升高,差异有统计学意义(P均<0.001)。Ja组的NF-κB p65亚基阳性染色率(15.5±4.3)明显高于SO组(5.1±1.9)(P<0.001)。而RT-PCR发现Ja组TNF-αmRNA相对表达量(1.3±0.4)明显高于SO组(0.8±0.1)(P<0.001)。结论 NF-κB在胆汁淤积早期呈高表达状态,其可能是调控胆汁淤积性肝损伤的重要通路,能为防止肝损伤提供治疗靶点。

To investigate the value of change of activity of NF-lcB during cholestasis injury induced by the obstruction of bile duct. Methods Twenty rats were divided randomly into two groups： Sham- operation group （SO group） and Jaundice control group （Ja group）. The rats in Ja group suffered the ligation of common bile duct. All the rats were sacrificed at 7 d post surgery. The ALT, TB and TBA were determined. The expression of NF-~~B p65 was assessed with immunohistological staining. The RT-PCR was employed to detect the TNF- ct mRNA level in liver samples. Results The ALT level in Ja group （145.7 ＋ 33.5） p mol/L was significantly higher than SO group （40.7 ＋ 7.3） lamol/L （P〈0.001）. Meanwhile, the leve of TBA and TBIL in Ja group were also significantly higher than SO group （both P〈0.001）. The positive staining rate of NF-~~B p65 subunit in Ja group showed significant higher value （15.5 ＋ 4.3） compared with SO group （5.1 ＋ 1.9） （P〈0.001）. The expression ratio of TNF-ct mRNA in Ja group was （1.3 ~ 0.4）, which was also significantly higher than that in SO group （0.8 ~ 0.1） （P〈0.001）. Conclusion The NF-~cB is up-regulated while cholestasis existed. The activity of NF-~： B might be the novel target for the treatment of cholestasis injury.