摘要
目的观察参附(Shenfu,SF)注射液预处理对大鼠肝移植缺血再灌注后Kupffer细胞中糖皮质激素受体(glucocorticoid receptor,GR)、核转录因子κB(NF-κB)表达的影响,探讨参附预处理保护缺血再灌注损害(ischemia reperfu-sion injury,IRI)的机制。方法将雄性SD大鼠按完全随机分组法分为正常对照组、缺血再灌注组和SF组,其中缺血再灌注组、SF组建立肝移植缺血再灌注模型,于缺血再灌注后6 h分离培养受体移植肝脏的Kupffer细胞。通过RT-PCR、Westernblot和激光共聚焦显微镜技术、免疫荧光法、ELISA等实验技术检测Kupffer细胞GR的mRNA和蛋白表达、NF-κB的蛋白表达、培养上清中TNF-α的含量。结果缺血再灌注组Kupffer细胞GR mRNA和蛋白表达均明显低于正常对照组(P<0.01),缺血再灌注组NF-κB、培养上清TNF-α表达量(830.19±71.34)明显高于正常对照组NF-κB、培养上清TNF-α表达量(65.41±37.63,P<0.01)。与缺血再灌注组相比,SF组GR水平明显上升(P<0.01),NF-κB活性、TNF-α表达量(543.59±41.27)明显下降(P<0.01)。结论缺血再灌注损伤会导致Kupffer细胞GR的表达下调,NF-κB的激活;参附预处理可以改善肝移植缺血再灌注后Kupffer细胞的激活。
Objective To observe the effect of pretreatment with Ginseng and Prepared Aconite Injection on expressions of glucocorticoid receptor（GR） and nuclear factor kappa B（NF-κB） in Kupfer cells（KCs） of rats after ischemia/reperfusion（I/R） injury due to liver transplantation.Methods Thirty male SD rats were randomly divided into control group,I/R group,and Ginseng and Prepared Aconite Injection group.An liver transplantation I/R injury model was established in 2 later groups.The injection at 2 ml/100 g was given by tail vein to the donors of the Injection group 1 h before operation.Six hours after I/R injury,KCs were isolated from cultured blood of liver transplantation recipients.Expressions of GR mRNA and protein as well as NF-κB in KCs were detected,and TNF-α level in supernatant was measured by RT-PCR,Western blotting,laser confocal microscopy and ELISA,respectively.Results The expression level of GR mRNA and protein in KCs was significantly lower,while the expression level of NF-κB in KCs and the TNF-α level in supernatant were significantly higher in I/R group than in control group（TNF-α level： 830.19±71.34 vs 65.41±37.63,P〈0.01）.The GR expression level,NF-κB activity,and TNF-α level in supernatant were significantly lower in Ginseng and Prepared Aconite Injection group than in I/R group（TNF-α level： 543.59±41.27 vs 830.19±71.34,P〈0.01）.Conclusion I/R injury can down-regulate the expression of GR and the activation of NF-κB in KCs.Pretreatment with Ginseng and Prepared Aconite Injection can improve the activation of KC after I/R injury due to liver transplantation.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2011年第12期1216-1219,共4页
Journal of Third Military Medical University
基金
国家自然科学基金(30772098)~~
关键词
KUPFFER细胞
肝移植
再灌注损伤
参附注射液
糖皮质激素受体
核因子-ΚB
大鼠
Kupffer cell
liver transplantation
reperfusion injury
Ginseng and Prepared Aconite Injection
glucocorticoid receptor
nuclear factor kappa B
rats
