大鼠缺氧/复氧皮质神经元高迁移率族蛋白B1和核因子κB的表达

Expression of HMGB1 and NF-κB in cultured neurons of rat after hypoxia/reoxygenation

目的:探讨高迁移率族蛋白B1(HMGB1)、核因子κB(NF-κB)对缺血再灌注损伤(IRI)中神经元凋亡的影响。方法:取新生SD大鼠的脑组织行原代大鼠皮质神经元培养,培养7d后分4组,A组为正常对照组(不予特殊处理),B组采用缺氧/复氧模拟IRI,C组单纯给予采用重组HMGB1(rHMGB1)刺激,D组给予NF-κB抑制剂吡咯烷二硫代氨基甲酸盐(PDTC)预处理+缺氧/复氧。4组在复氧或rHMGB1刺激结束后0、1、3、6、12、24h采用免疫细胞化学染色方法检测细胞中HMGB1、NF-κB的表达,MTT法检测细胞存活率,TUNEL法检测凋亡细胞。结果:①与A组比较,各时间点B、C组神经元HMGB1和NF-κB阳性表达增加,细胞存活率下降,神经元凋亡增多(P<0.05)。②与B组相比,D组神经元HMGB1和NF-κB表达减少,细胞存活率增加,凋亡减少(P<0.05)。③B组神经元HMGB1、NF-κB的表达与凋亡细胞数、HMGB1与NF-κB的表达均呈正相关(r分别为0.660、0.774、0.737,P均<0.05)。结论:IRI可使神经元HMGB1和NF-κB表达增加,2者相互诱生,共同参与了神经元的凋亡。

Aim： To explore the effects of HMGB1 and NF-κB on neuronal apoptosis caused by ischemia-repeffusion injury. Methods：The cortical neurons cultured for 7 days were randomly divided into A （normal control group） , B [ hypoxia/reoxygenation （H/R） alone] , C （treatment with rHMGB1 ,and without H/R） , D （pretreatment with PTDC and H/R） groups. The expressions of HMGB1, NF-κB were tested by immunoeytoehemical technique, eell survival rate was detected by MTT colorimetry, and the apoptotic was detected by TUNEL, after reoxygenation or after finishing treatment with rHMGB1 0,1,3, 6,12,24 h. Results：Compared with those of Group A, in Group B and C, the expressions of HMGB1 and NF-κB in neurons were stronger, and the apoptotie neuron was more obviously,the cell survival rate decreased（ P 〈 0.05 ）. Compared with those of Group B, the expressions of HMGB1 and NF-κB, and the apoptotie neurons quantity were lower and the survival rate increased in Group D （P 〈 0.05）. The expressions of HMGB1 and NF-κB were both positively correlated to the apoptosis, and HMGB1 had positive correlation with NF-κB in Group B（ r was 0. 660,0. 774,0. 737 ,respectively,P 〈 0.05）. Conclusion ：IRI could increase the expressions of HMGB1 and NF-κB of neurons. HMGB1 and NF-κB may be induced mutually in neurons,and related to neurons apoptosis.