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目的探讨NF-κB(核因子-κB)和PUMA(p53正向凋亡调节因子)在大鼠重症急性胰腺炎致急性肺损伤(SAP-ALI)发病中的作用及脯氨酸二硫代氨基甲酸酯(PDTC)对此过程的影响。方法 SD大鼠随机分为假手术组(A组),SAP-ALI组(B组),SAP+PDTC干预组(C组),每组24只。各组再按6,12,24 h时点分为3个亚组,每亚组8只。A组开腹后翻动胰腺数次;B组采用胰胆管逆行注入5%牛磺胆酸钠(1 mL/kg)诱导SAP-ALI模型;C组在B组的基础上于术前1 h给予PDTC(15 mg/kg),各组按各时点处死大鼠。观察胰腺和肺脏病理变化。检测不同组肺脏组织中NF-κBp65,PUMA和Caspase-3的表达及Caspase-3活性;检测组织TNF-α,MIP-2和ICAM-1 mRNA的表达、髓过氧化物酶(MPO)的活性和肺泡上皮细胞凋亡指数。结果成功建立大鼠SAP-ALI模型。Western-blotting和RT-PCR结果显示,B组肺上皮细胞NF-κB p65,PUMA和Caspase-3蛋白表达在12 h后显著增加(P<0.05),NF-κB p65与PUMA呈高度正相关(r=0.987,P<0.01)。TNF-α,MIP-2,ICAM-1mRNA表达及MPO和Caspase-3活性明显增加(P<0.01)。C组肺损伤病理组织学评分在术后各时点较B组显著下降(P<0.05),C组12 h后的肺组织NF-κB p65,PUMA和Caspase-3蛋白表达及MPO和Caspase-3活性明显降低(P<0.05);TNF-α,MIP-2,ICAM-1 mRNA表达明显下降(P<0.05),C组细胞凋亡指数较B组明显降低(P<0.01)。结论大鼠SAP-ALI既与NF-κB活化引起的炎症因子释放有关又与NF-κB活化引起的PUMA上调促进细胞凋亡有关。PDTC通过抑制NF-κB活化,下调炎症因子释放及NF-κB活化引起的PUMA表达可抑制肺组织的细胞凋亡,从而减轻SAP-ALI的程度。
Objective To investigate the role of NF-κB and PUMA in the pathogenesis of acute lung injury (SAP-ALI) induced by severe acute pancreatitis in rats and the effects of proline dithioamino Effect of Formate (PDTC) on this Process. Methods SD rats were randomly divided into sham operation group (A group), SAP-ALI group (B group) and SAP + PDTC intervention group (C group), 24 rats in each group. The groups were divided into 3 subgroups at 6, 12 and 24 h points, with 8 in each subgroup. In group A, the pancreas was overturned several times after open; in group B, SAP-ALI model was induced by retrograde injection of 5% sodium taurocholate (1 mL / kg) in group B; group C was given on the basis of group B PDTC (15 mg / kg), rats in each group were sacrificed at each time point. Observe the pathological changes of pancreas and lung. The expression of NF-κBp65, PUMA and Caspase-3 and the activity of Caspase-3 were detected in different groups of lung tissues. The expressions of TNF-α, MIP-2 and ICAM-1 mRNA and the activity of myeloperoxidase And alveolar epithelial cell apoptosis index. Results The SAP-ALI model was established successfully. The results of Western-blotting and RT-PCR showed that the expression of NF-κB p65, PUMA and Caspase-3 in group B were significantly increased after 12 h (P <0.05), and the positive correlation between NF-κB p65 and PUMA r = 0.987, P <0.01). TNF-α, MIP-2, ICAM-1mRNA and MPO and Caspase-3 activity increased significantly (P <0.01). The histopathological scores of lung injury in group C were significantly lower than those in group B at all time points after operation (P <0.05). The expression of NF-κB p65, PUMA and Caspase-3 in lung tissue and the expression of MPO and Caspase- (P <0.05). The expressions of TNF-α, MIP-2 and ICAM-1 mRNA were significantly decreased (P <0.05). The apoptosis index of group C was significantly lower than that of group B (P <0.01). Conclusion SAP-ALI is associated with the release of inflammatory cytokines induced by activation of NF-κB and the up-regulation of PUMA induced by NF-κB activation. PDTC can inhibit the apoptosis of lung tissue by inhibiting the activation of NF-κB, down-regulating the release of inflammatory cytokines and the activation of NF-κB, thereby reducing the level of SAP-ALI.