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-  2018 

抑制Kupffer细胞功能对梗阻性黄疸大鼠肝功能的保护作用及其机制 Protective Effect of Kupffer Cell Inhibition on Hepatic Function of Obstructive Jaundice Rats Through Nrf2/HO-1 Signaling Pathway

Keywords: Kupffer细胞,梗阻性黄疸,肝损害,抗氧化应激,Nrf2/HO-1通路

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Abstract:

目的:探讨抑制Kupffer细胞的功能对梗阻性黄疸大鼠肝脏Nrf2/HO-1通路的影响和保护肝功能的机制。方法:雄性Wistar大鼠60只,采用数字随机表法将大鼠平均分为6组:假手术组(SHAM组)、假手术+氯化钆处理组(SHAM+GdCl_3组)、梗阻性黄疸7d组(BDL-7D组)、梗阻性黄疸7d+氯化钆处理组(BDL-7D+GdCl_3组)、梗阻性黄疸14d组(BDL-14D)、梗阻性黄疸14d+氯化钆处理组(BDL-14D+GdCl_3组)。在梗阻性黄疸模型建立24h后由大鼠尾静脉注射氯化钆(GdCl_3,20mg/kg)。在相应时间点检测各组大鼠血清检测门冬氨酸氨基转移酶(AST)、总胆红素(TBIL)、碱性磷酸酶(ALP);肝组织匀浆检测丙二醛(MDA)、超氧化物歧化酶(SOD)含量;Western Blot及免疫组织化学法检测肝组织Nrf2、HO-1蛋白表达情况;肝组织HE染色观察病理变化。结果:AST、ALP、TBIL三个指标在SHAM组、BDL-7D组、BDL-14D组依次增高(P<0.05),SHAM组与SHAM+GdCl_3组未见明显差异;AST、TBIL两个指标在GdCl_3干预的梗阻性黄疸组要低于相应的梗阻性黄疸组(P<0.05),但ALP则未见明显差异。Western Blot结果显示Nrf2及HO-1的表达在SHAM组、BDL-7D组、BDL-14D组的依次增强,且差异均有统计学意义(P<0.05),在GdCl_3干预的梗阻性黄疸组要高于相应的梗阻性黄疸组(P<0.05),但SHAM+GdCl_3组与SHAM组未见明显差异。免疫组化染色结果也显示:SHAM组的Nrf2及HO-1蛋白均只有少量阳性表达。BDL组和BDL+GdCl_3组阳性表达细胞数均较SHAM组与SHAM+GdCl_3组明显增多(P<0.05),而且相对应的用氯化钆干预的大鼠肝脏Nrf2及HO-1表达要高于对照组。肝脏HE染色结果示:SHAM组和SHAM+GdCl_3组的肝组织未见异常;BDL-7D组肝细胞水肿,肝血窦扩张,肝细胞索排列紊乱,BDL-7D+GdCl_3组的表现和BDL-7D组的表现相似但程度较轻;BDL-14D组出现片状坏死灶,汇管区明显增生,而BDL-14D+GdCl_3组则表现为中央静脉周围肝细胞水肿,汇管区也明显增生。结论:抑制Kupffer细胞功能对梗阻性黄疸大鼠肝脏有保护作用,其机制可能是抑制炎症反应和上调抗氧化应激Nrf2/HO-1通路的蛋白表达来保护肝功能

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