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目的观察内质网应激相关因子氧调节蛋白150(ORP150)和C/EBP同源蛋白(CHOP)在缺氧缺血性脑损伤(HIBD)新生大鼠脑皮质内的表达,探讨内质网应激在新生大鼠HIBD中的作用。方法新生7日龄SD大鼠随机分为假手术组和缺氧缺血(HI)组。每组按照观测的时间点不同分为3 h、6 h、12 h、24 h、3 d、7 d 6个亚组,每个亚组8只。在每个时间点将大鼠断头后取皮质脑组织匀浆,用Western blot方法检测不同时间点其脑皮质ORP150及CHOP的动态变化。用原位末端标记法检测相应时间点光镜下其脑皮质组织的凋亡细胞数。结果 1.HI组CHOP表达水平在HIBD后各时间点均高于假手术组(P<0.05),且在24 h达到高峰;ORP150表达水平在完成HIBD后3 h开始增加,6 h达到高峰,后逐渐下降,7 d时与假手术组比较差异无统计学意义(P>0.05)。2.HI后3 h,HI组即发现结扎侧脑皮质有少量的凋亡细胞,随着HI时间的延长,凋亡细胞逐渐增多,24 h达到高峰后下降。3.CHOP的表达变化与神经细胞凋亡时间趋势呈正相关(r=0.911,P<0.05)。结论 HIBD诱发了内质网应激,可能通过上调ORP150表达参与启动未折叠蛋白反应过程,而通过上调CHOP表达诱导神经细胞凋亡的发生。
Objective To observe the expression of endoplasmic reticulum stress-related factors oxygen regulatory protein 150 (ORP150) and C / EBP homologous protein (CHOP) in the cerebral cortex of neonatal rats with hypoxic-ischemic brain damage (HIBD) The role of stress in neonatal rat HIBD. Methods Newborn 7-day-old SD rats were randomly divided into sham-operated group and hypoxic-ischemic group (HI). Each group was divided into 6 subgroups of 3 h, 6 h, 12 h, 24 h, 3 d, and 7 d according to the observed time points, and each group was divided into 8 subgroups. At each time point, the rats were decapitated and the homogenate of cortex was taken. The dynamic changes of ORP150 and CHOP in cerebral cortex at different time points were detected by Western blot. The number of apoptotic cells in cerebral cortex under light microscope at the corresponding time points was detected by in situ end labeling. The level of CHOP in HI group was higher than that in sham operation group at each time point after HIBD (P <0.05), and peaked at 24 h. The expression level of ORP150 increased at 3 h after HIBD and reached the peak at 6 h, Then gradually decreased. There was no significant difference between the sham group and the 7th day (P> 0.05). In HI group, a small amount of apoptotic cells were found in the cortex of the ligation side at 3 h after HI. As the HI time prolonged, the number of apoptotic cells increased gradually and reached the peak at 24 h. There was a positive correlation between the expression of CHOP and the trend of neuron apoptosis time (r = 0.911, P <0.05). Conclusion HIBD induces endoplasmic reticulum stress, which may be involved in initiating the process of unfolded protein by up-regulating ORP150 expression and inducing neuronal apoptosis by up-regulating CHOP expression.