To observe effect of acupuncture combined with hypothermia therapy on MAPK/ERK pathway and apoptosis related factorsin rats suffered cerebral ischemia reperfusion and to explore underlying mechanisms. Methods: Middle cerebral artery ischemia model were established.Ninety SD rats were randomly assigned into a blank group, a control group, a model group, an acupuncture group, a mild hypothermia group, and an acupuncture with hypothermia group. After 72 h treatment, nerve function defect scores were observed, and infarction area percent was detected by 2, 3, 5-triphenyl-2H-tetrazolium chloride (TTC) staining; expressions of Bcl-2 and Bax were examined by immunohistochemistry; apoptotic cells were detected by TUNEL assay; and expression levels of phospho-mitogen-activated protein kinase(p-MEK2) and phospho-extracellular signal regulated kinase 1/2 (p-ERK1/2) in the rats' hippocampus ischemic side were determined by Western blot. Results: In the rats of the model group, the neural function defect scores, the infarction area percent, the expression level of Bax, and apoptotic cells increased, while the level of Bcl-2 decreased significantly. The level of p-MEK2 and p-ERK1/2 increased obviously compared with the blank and control groups (P<0.05 or P<0.01). After treatment with acupuncture and hypothermia, the neural function defect scores, infarction area percent, and the level of Bax, apoptotic cells and the levels of p-MEK2 and p-ERK1/2 were significantly decreased, while the level of Bcl-2 in the treatment group was significantly elevated (P<0.05 or P<0.01) compared with the model group. Compared with the acupuncture group or the hypothermia group, the neural function defect scores and the levels of p-MEK2 and p-ERK1/2 in the acupuncture combined with hypothermia group were significantly reduced (P<0.05 or P<0.01). Conclusion: Acupuncture and hypothermia therapy can improve cerebral function, and reduce the cerebral injury through down-regulation of Bax level, and up-regulation of Bcl-2 level, which is related to reducing the levels of p-MEK2 and p-ERK1/2. The therapeutic effects on cerebral ischemia reperfusion injury for combination of acupuncture with hypothermia are better than those with single application of acupuncture or hypothermia.
目的：观察针刺联合亚低温对脑缺血再灌注大鼠脑组织MAPK/ERK通路及凋亡相关因子的影响，探讨针刺联合亚低温治疗缺血性中风的机制。方法：参照Longa 线拴法复制并改良大脑中动脉缺血模型，将90只SD大鼠随机分为空白组、假手术组、模型组、针刺组、亚低温组、针刺联合亚低温组，针刺及亚低温治疗72 h后，观察神经功能缺损评分，采用2, 3, 5-苯氯化四氮唑(2, 3, 5-triphenyl-2H-tetrazolium chloride，TTC)染色检测梗死面积百分比，免疫组织化学法检测Bcl-2及Bax表达水平，TUNEL法检测凋亡细胞，Western印迹检测大鼠缺血侧海马组织MEK-2，ERK1/2蛋白的磷酸化水平。结果：造模后，模型组大鼠神经功能缺损评分增加，梗死面积百分比升高，凋亡细胞及Bax表达增多，Bcl-2表达减少，MEK-2，ERK1/2蛋白的磷酸化水平明显增高，与空白组及假手术组比较差异有统计学意义(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、梗死面积百分比、Bax表达减少，凋亡细胞及MEK-2和ERK1/2蛋白的磷酸化水平均明显降低，Bcl-2表达升高，与模型组比较差异有统计学意义(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，MEK-2和ERK1/2蛋白的磷酸化水平下降(P<0.05或P<0.01)。与亚低温组比较，针刺联合亚低温组MEK-2和ERK1/2蛋白的磷酸化水平下降(P<0.01)。结论：针刺及亚低温均可改善神经功能缺损，减少脑梗死面积百分比、细胞凋亡、Bax表达及升高Bcl-2表达，对脑组织起保护作用。其机制可能是通过MAPK/ERK通路，降低MEK-2和ERK1/2蛋白的磷酸化水平及调节凋亡相关因子Bcl-2表达和Bax表达来实现的。针刺联合亚低温在降低神经功能评分及下调脑缺血再灌注损伤大鼠MEK-2，ERK1/2 蛋白的磷酸化水平上优于针刺或亚低温单独应用。.