内源性硫化氢对急性非结石性胆囊炎NF-κB的保护作用

摘要

目的 探讨急性非结石性胆囊炎(AAC)中NF-κB的表达及内源性硫化氢的保护作用.方法 采用胆总管结扎(BDL)方法构建AAC动物模型;40只成年豚鼠被随机分为正常组(normal组)、假手术对照组(sham组)、BDL-48 h组(BD-48 h组)和炔丙基甘氨酸(PAG)+BDL-48 h组(PAG组),每组10只.造模48 h后收集各组胆囊组织及动物血清.HE染色评价胆囊炎症并测定各组胆囊组织胱硫醚γ-裂解酶(CSE)活性;全自动生化分析仪检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)和胆红素(Bilirubin)水平;免疫组织化学法和Western blot法分别检测各组胆囊组织中CSE、NF-κB p65的表达.结果 PAG组炎症评分均明显高于BDL-48 h组、normal组和sham组,差异均有统计学意义(P<0.05);与normal组和sham组比较,BDL-48 h组和PAG组豚鼠血清ALT、AST、Bilirubin水平及胆囊组织CSE活性均明显增高(P<0.05),其中,PAG组CSE活性明显低于BDL-48 h组,差异有统计学意义(P<0.05);免疫组织化学法和Western blot法显示,PAG组胆囊的NF-κB p65蛋白水平均明显高于BDL-48 h组、normal组和sham组,差异均有统计学意义(P<0.05).结论 NF-κB的活化参与了急性非结石性胆囊炎的病理过程,内源性硫化氢可能通过抑制NF-κB的表达,从而减轻胆囊的炎症程度.%Objective To investigate the expression of nuclear factor-κB (NF-κB) in gallbladder during acute acalculous cholecystitis (AAC), and to study the protective effect of endogenous hydrogen sulfide (H2S) on NF-κB. Methods The model of AAC in guinea pig was established by common bile duct ligation (BDL), and 40 adult guinea pigs were randomly divided into normal group (normal group), sham control group (sham group), BDL for 48 hours group (BDL-48 h group), and DL-propargylglycine+BDL-48 h group (PAG group). Each group had 10 ani-mals. 48 hours after BDL, gallbladder tissues and serum of all animals were collected. Hematoxylin and eosin-stained slides of gallbladder samples were scored for inflammation, and cystathionineγ-lysae (CSE) activity in gallbladder tis-sue was examined. The level of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and bilirubin in se-rum were detected by full automatic biochemical analysis system. Immunohistochemistry and western blotting were used respectively to determine the expression levels of CSE and NF-κB p65. Results The inflammation score in PAG group was higher than those in normal, sham and BDL-48 h group (P<0.05). Compared with normal and sham group, the levels of ALT, AST, bilirubin and CSE activity were significantly increased in BDL-48 h and PAG group (P<0.05). And the CSE activity in PAG group was significantly lower than that in BDL-48 h group (P<0.05). Immuno-histochemistry analysis and western blotting showed that the level of NF-κB p65 protein in PAG group was signifi-cantly higher than those in normal, sham and BDL-48 h group (P<0.05). Conclusion Activation of NF-κB is in-volved in the pathological process of ACC. And Endogenous H2S may help reduce the inflammation in gallbladder by inhibiting expression of NF-κB.