高张溶液复苏大鼠创伤失血性休克对肺NF-κB活性影响的研究

Effects of Hypertonic Solution on Nuclear Translocation of Nuclear Factor kappa B in Lung following Trauma/Hemorrhagic Shock and Resuscitation

目的探讨高张溶液复苏创伤失血性休克(THS)模型大鼠对肺组织NF-κB细胞核内移位及i NOS活性、NO和TNF-α相关炎症介质生成量的影响。方法制作大鼠THS模型,分为假THS(Sham)组、THS乳酸林格氏液联合6%羟乙基淀粉复苏(LRH)组、THS 7.5%盐水/6%羟乙基淀粉复苏(HTH)组(每组n=8)。采用免疫组织化学技术观察肺组织细胞核NF-κB p65阳性数目和染色深度;Western blot法检测肺组织核蛋白中NF-κBp65含量,判定NF-κB细胞核内移位程度;化学显色法、硝酸还原酶法、ELISA法分别测定肺组织中i NOS活性、NO和TNF-α生成量。结果 HTH组肺组织NF-κB p65阳性的细胞核数量、染色深度及细胞核NF-κB p65蛋白含量均明显低于LRH组;i NOS活性、NO和TNF-α生成量低于LRH组。结论 HTH复苏THS模型大鼠可抑制肺组织中NF-κB蛋白向细胞核内移位和活化,抑制相关炎症介质i NOS的活性和NO、TNF-α的生成。

Objective Lung is an organ susceptible to be damages following trauma-hemorrhagic shock（THS） and resuscitation.It has been known that some factors such as NO,TNF-α,the production of which is controlled by nuclear factor kappa Binding protein（NF-κB） is under many pathological states,play crucial role for acute lung injury.Previous studies showed that resuscitation with hypertonic solution for THS could impact on inflammatory reactions and reduce organs damage,and study in vitro found that hypertonic solution could inhibit cytokine-induced nuclear factor-κB activation.However,the role of HTH resuscitating THS on NF-κB activity remains unknown.This research aimed to evaluate the effects of resuscitation with hypertonic solution on NF-κB nuclear translocation and production of TNF-α and NO2-/NO3-,the metabolites of NO,in rat model of THS.Methods Model of THS in rats was established.Rats suffered from THS were resuscitated with Ringer＇s lactate/6% Hydroethylstarch solution（LRH）,or 7.5% Sodium chloride/6% Hydroethylstarch solution（HTH）,or no THS and no resuscitation（Sham）.Immunohistochemistry and Western blotting were used to evaluate NF-κB nuclear translocation,ELISA to measure TNFa,and iNOS activity and NO2-/NO3-in lung tissues were also monitored.Results 24 hours after THS and resuscitation,the number of cells with positive nuclear NF-κB P65 staining,as well as the amount of NF-κB P65 in nucleus of lung tissues in rats resuscitated with HTH were significantly less compared to rats resuscitated with LRH;these were associated with a decrease of iNOS activity,and a lower production of NO2-/NO3-and TNFa in lungs.Conclusions These results indicated that rats with THS resusciated with a small volume of hypertonic solution reduced the early acute lung injuries by inhibiting NF-κB activation,which is directly responsible for TNF-a and iNOS activation leading to acute lung injury.