Effect of acupuncture plus mild hypothermia on MAPK/ERK pathway of brain tissues in rats with cerebral ischemia-reperfusion injury
Effect of acupuncture plus mild hypothermia on MAPK/ERK pathway of brain tissues in rats with cerebral ischemia-reperfusion injury
复制标题
针刺联合亚低温对脑缺血再灌注损伤大鼠脑组织MAPK/ERK通路的影响
DOI：
10.1007/s11726-016-0942-2
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发表时间：
2016
影响因子：
0.5
通讯作者：
Hao
中科院分区：
文献类型：
--
作者：
Yaping Lin;Qin Liu;Chu;Wen Chen;Heng Xiao;Qian;Hao
ObjectiveTo observe the protective effect of acupuncture plus mild hypothermia on brain tissues in rats with cerebral ischemia-reperfusion injury (CIRI), and the influence on protein expression levels of phosphorylated Raf-1, MEK-2 and ERK1/2 in the mitogen-activated protein kinase (MAPK)/extracellular regulated protein kinases (ERK) pathway, and to explore the mechanism of acupuncture plus mild hypothermia therapy for the ischemic stroke.MethodsNinety Sprague-Dawley (SD) rats were randomly divided into a blank control group, a sham operation group, a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group, 15 rats in each group. Except the rats in the blank control group, the remaining rats were used to prepare the middle cerebral artery occlusion (MCAO) models according to the modified occlusion method using lines, while only the occlusion lines were inserted without blocking the brain arteries of rats in the sham operation group. When the vital signs of rats were stable, rats in the blank control group did not receive any intervention; rats in the sham operation group and the model group received fastening without treatment; rats in the acupuncture group, the mild hypothermia group, and the acupuncture plus mild hypothermia group were treated with the corresponding therapeutic methods. 72 h later, observed neurologic injury score, evaluated infarction area ratio by 2,3,5-tripheyl tetrazolium chloride (TTC) staining, determined apoptosis by TUNEL assay, and measured the phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels in rat ischemic hippocampal tissues by Western blot assay.ResultsCompared with the blank control group and the sham operation group, after modeling, the neurologic injury score, infarction area ratio and apoptotic cells were increased, and phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels were significantly increased in the model group; the differences were statistically significant (P<0.05 or P<0.01). Compared with the model group, after acupuncture or mild hypothermia therapy, neurologic injury score and infarction area ratio were decreased; apoptotic cells and phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels were significantly decreased; the differences were statistically significant (P<0.05 or P<0.01). Compared with the acupuncture group, neurologic injury score and phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels were decreased in the acupuncture plus mild hypothermia group; differences between the groups were statistically significant (P<0.05 or P<0.01). Compared with the mild hypothermia group, phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels decreased in the acupuncture plus mild hypothermia group, and differences were statistically significant (P<0.01).ConclusionAcupuncture or mild hypothermia therapy can improve neurologic injury, reduce infarction area and apoptosis, which brought about protective effect on the brain tissues, in the MCAO model. The protective effect of acupuncture plus mild hypothermia group is the strongest. The mechanism may involve the MAPK/ERK pathway, by reducing the phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels.摘要目的观察针刺联合亚低温疗法对脑缺血再灌注大鼠脑组织的保护作用, 以及对丝裂原激活的蛋白激酶 (mitogen-activated protein kinase, MAPK)/细胞外调节蛋白激酶(extracellular regulated protein kinases, ERK)通路上磷酸化Raf-1、MEK-2、ERK1/2 蛋白表达水平的影响, 探讨针刺联合亚低温疗法干预缺血性中风的机制。方法将90 只Sprague-Dawley (SD)大鼠随机分为空白组、假手术组、模型组、针刺组、亚低温组和针刺联合亚低温组, 每 组15 只。随机挑选15 只为空白组, 其余大鼠根据线拴法并改良以复制大脑中动脉缺血(middle cerebral artery occlusion, MCAO)模型, 假手术组只插入线栓不阻断大脑中动脉。待大鼠生命体征稳定后, 空白组不做任何处理, 假手术组和模型组只捆绑不治疗, 针刺组、亚低温组、针刺联合亚低温组进行相应的处理。72 h 后, 观察神经功 能缺损评分, 氯化三苯基四氮唑(2,3,5-tripheyl tetrazolium chloride, TTC)染色检测梗死面积比, TUNEL法检测凋亡细 胞, Western blot 法检测大鼠缺血侧海马组织磷酸化Raf-1、MEK-2、ERK1/2 蛋白的表达水平。结果造模后, 模 型组大鼠神经功能缺损评分、梗死面积比值及凋亡细胞增多, 磷酸化Raf-1、MEK-2、ERK1/2 蛋白的表达水平明显 增高, 与空白组及假手术组差异有统计学意义(P<0.05 或P<0.01)。经针刺及亚低温治疗后, 各治疗组神经功能 缺损评分、梗死面积比值减少, 凋亡细胞及磷酸化Raf-1、MEK-2、ERK1/2 蛋白的表达水平均明显降低, 与模型组 差异均有统计学意义(P<0.05 或P<0.01)。与针刺组比较, 针刺联合亚低温组神经功能缺损评分, 磷酸化Raf-1 蛋 白激酶(Raf-1)、丝裂原活化蛋白激酶-2 (mitogen-activated protein kinase-2, MEK-2)、细胞外信号调节激酶1/2 (extracellular signal regulated kinase1/2, ERK1/2)蛋白的表达水平下降, 组间差异有统计学意义(P<0.05 或P<0.01)。 与亚低温组比较, 针刺联合亚低温组磷酸化Raf-1、MEK-2、ERK1/2 蛋白的表达水平下降, 组间差异有显著统计学 意义(P<0.01)。结论针刺及亚低温疗法均可改善MCAO 模型大鼠的神经功能缺损, 减少脑梗死面积及细胞凋亡, 对脑组织起保护作用, 并且针刺联合亚低温组的保护作用较强。其机制可能是通过MAPK/ERK 通路, 降低磷酸化 Raf-1、MEK-2、ERK1/2 蛋白的表达水平。