失血合并内毒素诱发多器官功能障碍综合征的分子病理机制研究

Potential molecular mechanism of multiple organ dysfunction syndrome induced by hemorrhage and endotoxin

目的 :探讨失血性休克合并内毒素血症致多器官功能障碍综合征 ( MODS)家兔可能的分子病理机制。方法 :采用失血合并内毒素损伤诱发 MODS家兔模型 ,用原位杂交 ( ISH)和凝胶电泳迁移率改变分析法( EMSA)分别检测肺泡巨噬细胞 ( PAM)以及肝枯否氏细胞 ( KC)中 IκΒ激酶 β( IKKβ) m RNA表达和核因子κB( NFкB)的活性 ;用酶联免疫吸附法 ( EL ISA)检测两种细胞培养上清液中肿瘤坏死因子α( TNFα)含量 ;同时进行动脉血气、血生化及肺、肝、小肠病理组织学检查。结果 :诱发 MODS家兔 PAM和 KC中 IKKβm RNA表达 ( 0 .15± 0 .0 3,0 .17± 0 .0 4)、NFκB活性 ( 1.49± 0 .30 ,1.72± 0 .36 )和上清液 TNFα的含量〔( 2 79.74± 2 5 .91) ng/ L 和 ( 30 0 .0 5± 30 .86 ) ng/ L〕均较正常动物〔IKKβ m RNA表达 0 .0 3± 0 .0 1和 0 .0 2±0 .0 1,NFκB活性 0 .2 5± 0 .0 6和 0 .31± 0 .10 ,TNFα含量 ( 137.33± 6 .0 9) ng/ L 和 ( 134.85± 12 .0 9) ng/ L〕明显增高 ( P均 <0 .0 1) ;血尿素氮 ( BU N)和丙氨酸转氨酶 ( AL T)均明显升高 ,氧分压下降 ,内脏组织出现明显炎症病理改变。结论 :IKKβ、NFκB、TNFα在休克合并内毒素诱发 MODS的炎症病理改变中有重要作用。

Objective:To investigate the possible molecular mechanism of multiple organ dysfunction syndrome(MODS) associated with hemorrhagic shock followed by resuscitation and endotoxin.Methods: A rabbit model of MODS after hemorrhagic shock followed by resuscitation and endotoxin was used in this study.The expression of IκB kinaseβ(IKKβ),the activity of nuclear factorkappa B(NFκB) in macrophage(PAM) and Kupffer cell(KC),the concentration of tumor necrosis factorα(TNFα) in the culture supernatant were measured by in situ hybridization(ISH),electrophoretic mobility shift assay(EMSA) and enzyme linked immunoadsorbent assay(ELISA),respectively.Then the blood gas,biochemistrical and pathological changes in lungs、liver and intestines were examined in each groups.Results:In the MODS group,the expression of IKKβ mRNA(0 15±0 03,0 17±0 04 ),the activity of NFκB(1 49±0 30, 1 72±0 36) and the levels of TNFα 〔(279 74±25 91 ) ng/L,(300 05±30 86 ) ng/L〕 in PAM and KCs were significantly higher than those of normal controls 〔IKKβ mRNA 0 03±0 01 and 0 02±0 01;NFκB 0 25±0 06 and 0 31±0 10,TNFα(137 33±6 09 ) ng/L and (134 85±12 09 ) ng/L, respectively,all P<0 01〕.Also,the contents of blood urea nitrogen(BUN),alanine aminotransferase(ALT) in plasma significantly increased,the arterial oxygen partial pressure decreased,and the severity of organ damages in lungs,liver as well as intestines increased following MODS.Conclusion:The IKKβ expression,NFκB activation and cytokine release may play important roles in the pathogenesis of acute lung injury and MODS.