摘要
目的观察新生大鼠缺氧缺血性脑损伤后脑组织的病理损伤改变、脑组织内质网应激相关因子激活转录因子6(ATF6)、肌醇需求因子1(IRE1)、双链RNA依赖的蛋白激酶样内质网激酶(PERK)的分布及表达变化。方法7 d龄新生大鼠分为假手术组(Sham组)和缺氧缺血性脑损伤组(HIBD组)。采用改良Rice法建立HIBD模型。对造模72 h时间点的脑组织进行苏木精-伊红(HE)染色观察大脑病理变化,尼氏染色观察皮质及海马区神经元损伤情况,采用免疫组化法检测各组大鼠大脑皮质及海马区ATF6、IRE1、PERK的表达情况。造模后6、24、72 h分别处死大鼠,采用Western blot法检测各时间点ATF6、IRE1、PERK蛋白表达情况。结果HIBD后72 h,HE染色显示Sham组未见神经细胞损伤,HIBD组可见脑组织神经细胞损伤,且主要损伤部位为脑皮层及海马区域。尼氏染色显示HIBD组皮质及海马区受损神经元比例均高于Sham组,差异有统计学意义(P<0.001)。免疫组织化学染色显示,与Sham组比较,HIBD组脑组织大脑皮质及海马区中ATF6、IRE1、PERK阳性细胞数(棕黄色)增加(P<0.001)。HIBD组神经细胞胞质及核内ATF6表达均增加,胞核内IRE1及胞质内PERK表达增加。HIBD组ATF6、IRE1、PERK各时间点的蛋白相对表达水平均高于Sham组,差异有统计学意义(P<0.05)。结论新生大鼠缺氧缺血性脑损伤后,脑损伤的部位主要在大脑皮质及海马区,且大脑皮质及海马区组织内质网应激因子表达升高,提示内质网应激相关因子参与其病理性损伤。
Objective To observe the pathological changes of brain tissue in neonatal rats after hypoxic-ischemic brain damage(HIBD),the distribution and expression changes of endoplasmic reticulum stress(ERS)related factors includingactivated transcription factor 6(ATF6),inositol demand factor 1(IRE1)and double-stranded RNA-activated protein kinase-likeendoplasmic reticulum kinase(PERK)in brain tissue.Methods Seven-day-old newborn rats were divided into sham operation group(Sham group)and hypoxic-ischemic brain damage group(HIBD group).The modified rice method was used to establish the HIBD model.Hematoxylin-eosin(HE)staining was performed on the brain tissue at the 72 h point of modeling to observe the pathological changes of the brain,Nissl staining was used to observe the neuron damage in the cortex and hippocampus,and immunohistochemistry was used to detect the expression of ATF6,IRE1 and PERK in the cerebral cortex and hippocampus of rats in each group.The models were sacrificed at 6 h,24 h,and 72 h respectively,Western blot was used to detect the protein expression of ATF6,IRE1,and PERK at each time point at 6 h,24 h,and 72 h.Results 72 hours after HIBD,HE staining showed that no nerve cell injury was observed in the Sham group,while nerve cell injury was observed in the brain tissue of the HIBD group,and the main injury sites were the cerebral cortex and hippocampus.Nissl staining showed that the proportion of damaged neurons in the cortex and hippocampus of the HIBD group was higher than that in the Sham group,and the difference was statistically significant(P<0.001).Immunohistochemical staining showed that compared with the Sham group,the number of ATF6,IRE1,and PERK positive cells(brown-yellow)in the cerebral cortex and hippocampus of the HIBD group increased(P<0.001).In the HIBD group,the expression of ATF6 in the cytoplasm and nucleus of nerve cells increased,and the expression of IRE1 in the nucleus and PERK in the cytoplasm increased.The relative protein expression levels of ATF6,IRE1,and PERK at each time poi
作者
于海侠
刘文强
吴铭
王军
Yu Haixia;Liu Wenqiang;Wu Ming;Wang Jun(Dept of Pediatrics,Affiliated Hospital of Xuzhou Medical University,Xuzhou 221002;Dept of Radiology,Xuzhou Central Hospital,Xuzhou 221009)
出处
《安徽医科大学学报》
CAS
北大核心
2022年第9期1475-1480,共6页
Acta Universitatis Medicinalis Anhui
基金
江苏省卫生健康委科研项目(编号:H2019004)
徐州市科技项目(编号:KC18216)。
