沈　菁,雷晓明,宋　洋,谭　杏,刘　琴,戴丽雯,李文慧,郁　洁*（.湖南中医药大学针灸推拿学院，湖南长沙4008;.湖南中医药大学教务处，湖南长沙4008）



电针内关、百会穴对脑缺血/再灌注大鼠GRP78和Caspase-12基因表达的影响

沈菁1,雷晓明2,宋洋1,谭杏1,刘琴1,戴丽雯1,李文慧1,郁洁1*
（1.湖南中医药大学针灸推拿学院，湖南长沙410208;2.湖南中医药大学教务处，湖南长沙410208）

〔摘要〕目的观察电针内关、百会穴对脑缺血/再灌注（Ischemia/Reperfusion，I/R）大鼠脑GRP78和Caspase-12基因表达的影响，探讨针刺发挥脑保护作用是否与内质网应激凋亡通路有关。方法100只大鼠随机分为5组，每组20只，即正常组、假手术组、手术造模组、依达拉奉组和针刺干预组。采用线栓法建立大鼠大脑中动脉阻塞（middle cerebral artery occlusion，MCAO）脑I/R模型。采用TUNEL染色法，检测大鼠神经细胞凋亡指数；采用实时定量多聚酶链式反应（Real-Time polymerase chain reaction，RT-PCR），检测GRP78、Caspase-12 mRNA表达。结果与正常组和假手术组相比，手术造模组、依达拉奉组和针刺干预组大鼠细胞凋亡指数升高，GRP78和Caspase-12 mRNA表达增多，差异具有统计学意义（P＜0.05或P＜0.01）；与手术造模组相比，依达拉奉组和针刺干预组大鼠细胞凋亡指数和Caspase-12 mRNA表达均明显降低（P＜0.01），GRP78 mRNA表达明显升高（P＜0.01）；与依达拉奉组相比，针刺干预组大鼠各项指标差异无统计学意义（P＞0.05）。结论针刺内关、百会穴可以有效抑制脑缺血神经元细胞凋亡，其保护机制可能上调内质网应激保护性GRP78表达，同时抑制促凋亡Caspase-12表达有关。

〔关键词〕细胞凋亡;电针;脑缺血/再灌注损伤;百会;内关;GRP78;Caspase-12

Effect of Electroacupuncture at Neiguan and Baihui Points on GRP78 and Caspase-12 Gene Expression in Rats with Ischemia-Reperfusion Injury

SHEN Jing1, LEI Xiaoming2, SONG Yang1, TAN Xing1, LIU Qin1, DAI Liwen1,
Li Wenhui1, YU Jie1*
（1. Acupuncture and Tuina Massage School, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China；2. Academic Affairs Division, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China）

〔Abstract〕Objective To observe the effects of electro-acupuncture（EA）at Neiguan and Baihui point on GRP78 and Caspase-12 gene expression in rats with ischemia/ reperfusion（IR）injury and investigate protective effects of acupuncture wheather related to endocytoplasmic reticulum（ER）stressapoptosis passage. Methods 100 rats were randomly assigned to five groups, 20 in each group: the normal group, sham-operation group, operation model group, edaravone group and EA group. The IR model of middle cerebral artery occlusion（MCAO）was established by suture embolic method. The apoptosis index of nerve cells in rats were measured by TUNEL staining method. The mRNA expression of GRP78 and Caspase-12 were measure by Real-time polymerase chain reaction（RT-PCR）. Results Compared with normal group and sham-operation group, the apoptosis indexes and mRNA expression of GRP78 and Caspase-12 in operation group, Edaravone group and EA group were increased, with statistical significance（P＜0.05 or P＜0.01）; Compared with operation model group, the apoptosis indexes and Caspase-12 mRNA expression in edaravone group and EA group were decreased（P＜0.01）, GRP78 mRNA expression were increased obviously（P＜0.01）; there were no significant difference between edaravone group and EA group on the above indexes（P＞0.05）. Conclusion Acupuncture at Neiguan and Baihui points could effectively suppress the nerve cell apoptosis in cerebral ischemia. The underlying mechanism might be related to upregulation of the ERS-protective GRP78 expression and inhibition of apoptosis-promotion Caspase-12 expression.

〔Keywords〕cell apoptosis, electroacupuncture, cerebral ischemia/reperfusion injury, Baihui point; Neiguan point; GRP78, Caspase-12

近年来有关内质网应激介导凋亡通路的研究受到广泛关注，已发现内质网应激与许多疾病的发生发展有关，比如糖尿病、肿瘤等。……