CDK4-p RB-E2F1通路在创伤后应激障碍大鼠杏仁核神经细胞凋亡中的作用

Role of the CDK4-pRB-E2F1 pathway in apoptosis of amygdala neurons in rats with post-traumatic stress disorder

目的探讨CDK4-pRB-E2F1通路在创伤后应激障碍(PTSD)大鼠杏仁核神经细胞凋亡中的作用。方法选取健康雄性成年Wistar大鼠50只,采用单一连续应激(SPS)制备PTSD大鼠模型,按照SPS处理后1 d、4 d、7 d、14 d将大鼠随机分为4组,将未做任何处理大鼠作为对照组,每组10只。采用旷场实验和僵立行为测定动物行为学变化;Western blotting检测各组大鼠杏仁核神经细胞中pRB、E2F1蛋白表达;实时PCR检测各组大鼠杏仁核神经细胞中RB、E2F1 mRNA的表达。结果与对照组比较,SPS 7 d组大鼠进入旷场箱内中心区域较少,而僵立行为比例显著增高(均P<0.05)。与对照组比较,SPS 7 d组、SPS 14 d组大鼠杏仁核神经细胞pRB、E2F1蛋白表达增高(P<0.05);SPS 4 d组、SPS 7 d组、SPS 14 d组大鼠杏仁核神经细胞RB mRNA、E2F1 mRNA表达显著增高(P<0.05)。结论PTSD大鼠杏仁核神经细胞中pRB及E2F1高表达,CDK4-pRB-E2F1通路可能参与了PTSD杏仁核神经细胞的凋亡。

Objective To investigate the role of the CDK4-pRB-E2 F1 pathway in apoptosis of amygdala neurons in rats with posttraumatic stress disorder(PTSD).Methods Fifty healthy male adult Wistar rats were selected to establish a PTSD rat model by singleprolonged stress(SPS).The rats that were treated were randomly divided into four groups according to 1 d,4 d,7 d,and 14 d after SPS treatment.The rats that were not treated were included in the control group.Ten rats were included in each group.Open-field experiments and rigid behavior were used to assess changes in animal behavior;Western blotting was used to detect the protein expression of pRB and E2 F1 in the amygdala;and real-time polymerase chain reaction was used to detect the expression of RB mRNA and E2 F1 mRNA in amygdala neurons.Results Compared to the percentage of rats that entered the central area of the open-field box in the control group,that in the SPS 7 d group was decreased,and the proportion of rigid behavior had significantly increased in the SPS 7 d group than in the control group(P<0.05).Compared to the protein expression of pRB and E2 F1 in amygdala neurons of rats in the control group,that in amygdala neurons of rats in the SPS 7 d group and the SPS 14 d group was significantly increased(P<0.05).The expression of RB m RNA and E2 F1 mRNA in the amygdala of rats in the SPS 4 d group,SPS 7 d group,and SPS 14 d group was significantly increased compared with the control group(P<0.05).Conclusion pRB and E2 F1 are highly expressed in amygdala neurons of rats with PTSD.The CDK4-p RB-E2 F1 pathway may be involved in apoptosis of PTSD-affected amygdala neurons.