基于PERK-eIF2α信号通路探讨眼针对脑缺血再灌注损伤大鼠脑组织Atg12表达的影响

Effect of Eye Needle on Expression of Atg12 in Brain Tissue of CIRI Rats Based on PERK-eIF2αSignaling Pathway

目的 通过检测各组脑组织中磷酸化蛋白激酶R样内质网激酶(phosphorylation protein kinase-like ER kinase, p-PERK)、磷酸化真核起始因子2α(phosphorylated α subunit of eukaryotic initiation factor 2,p-eIF2α)、自噬相关基因12(antigen, Atg12)蛋白表达水平的变化,探讨眼针是否能够通过调控内质网应激信号通路PERK-eIF2α影响脑缺血再灌注损伤(cerebral ischemia reperfusion injury, CIRI)大鼠脑组织Atg12的表达。方法 建立大鼠脑缺血再灌注损伤模型,实验分为4组,即:对照组、假手术组、模型组、眼针组。末次针刺后,对各组大鼠进行神经功能缺损评分,透射电镜观察各组大鼠自噬小体的表达情况,然后采用Western blot法检测脑组织中p-PERK、p-eIF2α、Atg12蛋白表达。结果 与对照组和假手术组相比,模型组大鼠神经功能缺损评分、脑组织中p-PERK、p-eIF2α、Atg12蛋白表达均升高,差异具有统计学意义(P<0.01),自噬小体数量增多;与模型组相比,眼针组大鼠神经功能缺损评分、脑缺血组织中p-PERK、p-eIF2α、Atg12的蛋白表达明显降低,差异具有统计学意义(P<0.01),自噬小体数量减少。结论 眼针能够降低脑缺血再灌注损伤模型大鼠脑组织自噬蛋白Atg12的表达,其机制可能与调控内质网应激通路PERK-eIF2α有关。

Objective By detecting the expression levels of phosphorylation protein kinase-like ER kinase(p-PERK),phos-phorylatedαsubunit of eukaryotic initiation factor 2(p-eIF2α)and antigen(Atg)12 proteins in brain tissue of rats with cerebral ischemia and reperfusion injury(CIRI),it was investigated whether the eye needle affected the expression of Atg12 in brain tissue by regulating the PERK-eIF2αsignaling pathway of endoplasmic reticulum stress.s Methods The rat CIRI model was estab-lished,and the experiment was divided into four groups,namely:control group,sham surgery group,model group,and eye acu-puncture group.After the last acupuncture,neurological deficit scores were scored on each group,and the expressions of autoph-agtosomes in each group were observed by transmission electron microscopy,and then the expressions of p-PERK,p-eIF2αand Atg12 proteins in brain tissues were detected by Western blot method.s Results Compared with those of the control group and the sham surgery group,the expressions of p-PERK,p-eIF2αand Atg12 proteins in the brain tissues of rats in the model group were increased,the number of autophagosomes decreased.and the difference was statistically significant(P<0.01).Conclusions Eye acupuncture can reduce the expression of autophagy protein Atg12 in brain tissue of CIRI rats,and its mechanism may be re-lated to the regulation of endoplasmic reticulum stress pathway PERK-eIF2α.