急性胆汁淤积伴肝缺血大鼠一氧化氮和诱导型一氧化氮合酶表达下调

Verónica Barón, José Hernández, Martha Noyola, Bruno Escalante, Pablo Muriel
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引用次数: 17

摘要

胆汁淤积导致肝血流减少,有证据表明NO调节肝脏微血管灌注。因此,本研究的目的是评估胆汁淤积症中NO的合成。雄性Wistar大鼠胆管结扎(BDL)诱导胆汁淤积。测定血清中胆红素和酶活性。测定肝脏脂质过氧化、GSH、GSSG、糖原含量。进行组织病理学分析。采用Gries反应测定血清NO2−+NO3−浓度。采用iNOS多克隆抗体进行iNOS免疫印迹分析。7 d后BDL脂质过氧化升高,GSH/GSSG比值降低。组织病理学分析显示,大鼠血清NO2 - +NO3 -和肝脏iNOS蛋白减少,并伴有缺血。GSH通过增加iNOS mRNA水平和iNOS活性上调NO合成,因此GSH/GSSG比值的降低可能是iNOS蛋白和NO合成下调的原因,这可能解释了其他研究报道的肝缺血和胆汁淤积时肝血灌注减少的原因。
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Nitric oxide and inducible nitric oxide synthase expression are downregulated in acute cholestasis in the rat accompanied by liver ischemia

Hepatic blood flow decreases under cholestasis and there is evidence that NO regulates liver microvascular perfusion. Thus, the aim of the present study was to evaluate NO synthesis in cholestasis. Cholestasis was induced by bile-duct ligation (BDL) in male Wistar rats. Bilirubins and enzyme activities were measured in serum. Lipid peroxidation, GSH, GSSG and glycogen were determined in liver. Histopathological analysis was performed. Serum NO2+NO3 concentration was measured by the Gries reaction. iNOS immunoblot analysis was carried out using an iNOS polyclonal antibody. After 7 days of BDL lipid peroxidation increased while GSH/GSSG ratio decreased. Serum NO2+NO3 and liver iNOS protein were reduced, accompanied by ischemia as revealed by the histopathological analysis. GSH upregulates NO synthesis by increasing iNOS mRNA levels and iNOS activity, thus the reduction of GSH/GSSG ratio may be responsible for the downregulation of iNOS protein and NO synthesis, which in turn may explain the observed ischemia and the decreased hepatic blood perfusion in cholestasis reported by others.

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