CAJ | 학술논문

目的探讨糖皮质激素受体(GR)、核因子-κB(NF-κB)在创伤失血性休克后肝组织中的变化、相互关系,及其对肝损伤的作用机制.方法雄性健康Wistar大鼠96只,采用双侧股骨骨折伴失血性休克创伤模型,随机分成正常对照组6只,创伤休克组30只,GR阻断伴创伤休克组30只,NF-κB抑制伴创伤休克组30只.动态观察伤后0.5、2、4、6、8 h大鼠肝组织GR、NF-κB,肝脏病理,肝功能,血清TNF -α、IL-6等变化.GR采用免疫印迹法测定蛋白含量,NF-κB采用EMSA法测定结合活性,并进行计算机图像分析.结果肝组织GR的蛋白含量在创伤失血性休克后2 h即开始下降,4 h明显低于正常对照(P<0.01),6 h降至最低,8 h仍显著低于正常(P<0.01);NF-κB的活性伤后迅速升高,伤后6 h达到高峰(P<0.01).光镜下伤后4～8 h肝窦内少许淤血,有散在炎性细胞浸润;血清TNF-α、IL-6、ALT、TB伤后4 h 开始增高.GR阻断后再致伤,NF-κB在伤后各个时相点的表达均较未阻断有明显增高,光镜下伤后2 h肝窦内即可见较多炎性细胞浸润,血清TNF-α、IL-6、ALT、TB在伤后2 h即有明显升高(P<0.01).抑制NF-κB再致伤后,GR在伤后肝组织中的表达增强,TNF-α、IL-6伤后各个时相点均迅速回落,光镜下伤后4～8 h肝细胞变性明显好转,肝窦内见淤血减轻,仅见少许淋巴细胞及中性粒细胞浸润;伤后4 h,血清ALT、TB即明显下降.结论 GR、NF-κB参与了严重创伤失血性休克后肝损伤的发生,阻断GR使NF-κB的表达增强,肝损害程度加重;抑制NF-κB使GR表达增加,肝损害程度减轻.提示GR及NF-κB在严重创伤休克后肝组织细胞损伤过程中关系密切并起着重要作用.
목적 탐토당피질격소수체(GR)、핵인자-κB(NF-κB)재창상실혈성휴극후간조직중적변화、상호관계,급기대간손상적작용궤제.방법 웅성건강Wistar대서96지,채용쌍측고골골절반실혈성휴극창상모형,수궤분성정상대조조6지,창상휴극조30지,GR조단반창상휴극조30지,NF-κB억제반창상휴극조30지.동태관찰상후0.5、2、4、6、8 h대서간조직GR、NF-κB,간장병리,간공능,혈청TNF -α、IL-6등변화.GR채용면역인적법측정단백함량,NF-κB채용EMSA법측정결합활성,병진행계산궤도상분석.결과 간조직GR적단백함량재창상실혈성휴극후2 h즉개시하강,4 h명현저우정상대조(P<0.01),6 h강지최저,8 h잉현저저우정상(P<0.01);NF-κB적활성상후신속승고,상후6 h체도고봉(P<0.01).광경하상후4～8 h간두내소허어혈,유산재염성세포침윤;혈청TNF-α、IL-6、ALT、TB상후4 h 개시증고.GR조단후재치상,NF-κB재상후각개시상점적표체균교미조단유명현증고,광경하상후2 h간두내즉가견교다염성세포침윤,혈청TNF-α、IL-6、ALT、TB재상후2 h즉유명현승고(P<0.01).억제NF-κB재치상후,GR재상후간조직중적표체증강,TNF-α、IL-6상후각개시상점균신속회락,광경하상후4～8 h간세포변성명현호전,간두내견어혈감경,부견소허림파세포급중성립세포침윤;상후4 h,혈청ALT、TB즉명현하강.결론 GR、NF-κB삼여료엄중창상실혈성휴극후간손상적발생,조단GR사NF-κB적표체증강,간손해정도가중;억제NF-κB사GR표체증가,간손해정도감경.제시GR급NF-κB재엄중창상휴극후간조직세포손상과정중관계밀절병기착중요작용.
Objective To investigate changes and functions of glucocorticoid receptor(GR) and nuclear factor kappa B(NF-κB) in a rodent model of hepatic injury after trauma hemorrhagic shock. Methods Trauma hemorrhagic shock was produced by inducing bilateral femoral fractures in male Wistar rats. Ru486 was used to block GR expression, as well as Pyrrolidine dithiocarbamate(PDTC)was used to inhibit NF-κB activation 1 hour before induction. A total of 96 adult male Wistar rats were randomly divided into 4 groups: control group(n=6) , trauma group(n=30), block GR group, and inhibit NF-κB group (n=30). Measurements of hepatic GR and NF-κB, hepatic function markers, TNF-α and IL-6 were obtained at 0.5, 2, 4, 6, 8 h after trauma. Histopathological changes in liver tissues were also noted. Hepatic expression of GR was assayed by western blot, while NF-κB was determined by electrophoretic mobility shift assay and analyzed with computer imaging system. Results In rats with trauma shock, Protein content of GR decreased and NF- B increased significantly compared to the control group. GR reduced to the lowest, NF-κB added to highest at 6h after trauma in hemorrhagic shock after trauma. Hepatic congestion was minimal, there were little inflammatory cells infiltrated in hepatic sinusoid at 4～8 h after trauma. TNF-α, IL-6, ALT and TB increased at 4 h after trauma. After usage of GR blocking agent, expression of NF-κB obviously enhanced, there were much inflammatory cells infiltrated in hepatic sinusoid at 2 h after trauma. TNF-α, IL-6, ALT and TB obviously increased. After inhibiting the activity of NF-κB, protein content of GR gradually increased than trauma shock group. The contents of TNF-α, IL-6, ALT and TB were droped. Hepatic congestion was lessen, hepatic cell degeneration was improved. A little homeocyte and neutrophile granulocyte were obversed in sinus hepaticus. Conclusion GR and NF-κB might participate hepatic injury after trauma with hemorrhagic shock. GR blockage may increase expression of NF-κB, and aggravate liver injury. NF-κB inhibition may increase expression of GR, and relieve liver injury. It is indicated that GR and NF-κB act as important role in injury mechanism of hepatic tissue cell after trauma with hemorrhagic shock.