AMP活化蛋白激酶在大鼠缺血缺氧性脑损伤中的作用

Role of AMP-activated protein kinase in hypoxic-ischemic brain damage in rats

目的研究缺血缺氧性脑损伤(HIBD)大鼠模型中AMP活化蛋白激酶(AMPK)对脑组织和神经元的作用。方法本研究采用颈动脉分离结扎法建立缺血缺氧脑损伤大鼠模型,分为假手术组(sham组)、缺血缺氧脑损伤6 h模型组(HI6 h组)、缺血缺氧脑损伤12 h模型组(HI12 h组),以及等剂量compound C处理的假手术组(sham+CC组)、HI6 h组(HI6 h+CC组)和HI12 h组(HI12 h+CC组)。采用干湿重称量法计算大鼠脑含水量,TUNEL染色法检测大鼠脑组织神经元的凋亡情况,TTC染色法检测大鼠脑梗死体积,Autophagy Assay检测细胞的自噬,蛋白质免疫印迹法检测AMPK和p-AMPK蛋白的表达。结果与sham组相比,HI6 h组和HI12 h组大鼠的神经元凋亡数目显著增加(P<0.05),脑含水量显著增多(P<0.05),脑梗死体积显著增大(P<0.05)。与sham组相比,HI6 h组和HI12 h组大鼠中p-AMPK/AMPK相对表达量均升高,差异具有统计学意义(P<0.05)。与sham组相比,HI6 h组和HI12 h组大鼠中细胞自噬显著增加(P<0.05)。使用AMPK抑制剂Compound C处理后,HI12 h+CC组大鼠中p-AMPK/AMPK相对表达量降低,细胞自噬情况明显减少(P<0.01)。结论AMPK的激活在缺血缺氧脑损伤疾病中具有保护作用,可作为缺血缺氧脑损伤疾病的治疗靶标。

Objective To explore the effect of AMP-activated protein kinase(AMPK)on brain tissue and neurons in a rat model of ischemic-hypoxic brain damage(HIBD).Methods The carotid artery separation and ligation was used to establish ischemic and hypoxic brain injury rat models.The rats were divided into sham group,6 h model group(HI6 h group),12 h model group(HI12 h group),and sham+CC group,HI6 h+CC group and HI12 h+CC group.The rats were treated with the same dose of compound C in sham+CC group,HI6 h+CC group and HI12 h+CC group.Dry and wet weights were used to calculate the brain water content in rats.TUNEL staining was used to detect the apoptosis of neurons in rat brain tissue.TTC staining was used to detect the volume of cerebral infarction in rats.Autophagy assay was used for autophagy testing in rats.The protein expression of AMPK and p-AMPK was detected by Western blotting.Results Compared with sham group,the number of neuronal necrosis,the brain water content and the volume of cerebral infarction were increased significantly in HI6 h group and HI12 h group(P<0.05).Compared with sham group,the relative expression of p-AMPK/AMPK was significantly increased in HI6 h group and HI12 h group(P<0.05).Compared with sham group,the autophagy was significantly increased in HI6 h group and HI12 h group.However,the relative expression of p-AMPK/AMPK was decreased and the autophagy was also significantly reduced in HI12 h group after treated with AMPK inhibitor(compound C)(P<0.01).Conclusion The activation of AMPK has a protective effect against ischemic and hypoxic brain injury,which can be used as a therapeutic target for ischemic hypoxic brain injury disease.