唐冕 章琪 陈红霞

摘要 目的：評价针刺曲池穴及足三里穴对MCAO模型大鼠运动功能的影响及观察脑梗死灶周围组织PI3K/AKT信号通路表达的变化。方法：将45只SD大鼠随机分为假手术组、模型组及干预组，各15只。假手术组仅予切开皮肤分剥离神经血管后缝合，模型组及干预组大鼠予线栓法进行MCAO模型制备，造模成功后干预组大鼠予针刺曲池穴及足三里穴，1次/d，30 min/次，假手术组及模型组大鼠仅模仿捉拿、回笼，连续干预14 d，用Homecage Scan行为学评价大鼠不同时间段运动能力的变化，免疫组化检测PI3K、Bax的表达差异，免疫蛋白印迹（Western blotting）观察AKT、p-Akt的浓度变化。结果：1）Homecage Scan行为学检测显示：假手术组大鼠未见明显异常，造模大鼠出现明显偏瘫，随着时间的推移模型组及干预组大鼠花费在行走、后肢站立、舔毛3种行为的时间逐渐延长，其中干预组改善得更为明显（P<0.05）。2）造模组大鼠脑组织TTC染色出现明显白色梗死区域，干预组大鼠梗死区域明显小于模型组。3）免疫组化显示造模后大鼠PI3K降低，Bax升高，针刺足三里穴及曲池穴可上调PI3K及AKT的磷酸化程度，抑制了Bax的表达;4）Western blotting显示针刺足三里、曲池穴可上调造模大鼠AKT的磷酸化程度。结论：针刺曲池穴、足三里穴可改善脑缺血大鼠的神经缺损症状，其作用机制可能与PI3K/AKT信号通路有关。

关键词 MCAO模型;大鼠;曲池穴;足三里穴;PI3K/AKT信号通路

Effects of Acupuncture at Quchi （LI 11） and Zusanli （ST 36） on the Expression of PI3K/AKT Signal Pathway

in the Infarction Surrounding Tissue of MCAO Model Rats

Tang Mian1， Zhang Qi2， Chen Hongxia3

（1 General Medicine， School of Clinical Medicine， Hubei University of Science and Technology， Xianning 437100， Chia; 2 Traditional

Chinese Medicine Rehabilitation of Second Affiliated Hospital of Hubei University， Xianning 437100， China; 3 Department of

Pathophysiology， School of Basic Medical Sciences， Hubei University of Science and Technology， Xianning 437100， China）

Abstract Objective：To evaluate the effects of the acupuncture at Quchi （LI 11） and Zusanli （ST 36） on motor function of the MCAO model rats and to observe PI3K/AKT signal pathway expression changes in the surrounding tissue of cerebral infarcts. Methods：A total of 45 SD rats were randomly divided into blank group， model group and intervention group， with 15 in each group. Blank group received only the skin incision and sutured after vascular decollement， and model group and intervention group rats accepted string inserting method for MCAO model preparation. After the successful modeling， the rats of the intervention group acupunctured at LI 11 and ST 36， 1 time/day， 30 min/time. The rats of the blank group and the model group only imitate the arrest， and go back to the cages for 14 days continuous intervention. The athletic ability changes of rats in different periods by using Homecage Scan behavior were evaluated， and the expression differences of PI3K and Bax were detected by immunohistochemical. The concentration changes of AKT and p-AKT were observed by Western blotting. Results：1） Homecage Scan behavior test showed：the blank group rats did not see obvious abnormity， and the model rats appeared obvious hemiplegia. With the pass of time， rats in the model group and intervention group spent gradually extend time in walking， standing， licking the hairs， and the behavior of intervention group improved more significantly （P< 0.05）. 2） TTC staining of the brain tissue of rats in model group showed obvious white infarction area. The infarction area in the intervention group was significantly smaller than that in the model group. 3） Immunohistochemical revealed PI3K reduced and Bax increased after the model made， and acupunctured in Quchi （LI 11） and Zusanli （ST 36） could increase extent of phosphorylation of PI3K and AKT， and inhibit the expression of Bax. 4） Western blotting showed that acupuncture at Quchi （LI 11） and Zusanli （ST 36） could up-regulate AKT phosphorylation of modeling rats. Conclusion：Acupuncture at Quchi （LI 11） and Zusanli （ST 36） can improve the neurological defect symptoms of cerebral ischemia rats， whose the mechanism may be associated with PI3K/AKT signal pathway.

Key Words MCAO model rats， Quchi （LI 11）， Zusanli（ST 36）， PI3K/AKT signaling pathway

中图分类号：R255.2文献标识码：Adoi：10.3969/j.issn.1673-7202.2018.03.041

脑卒中以高发病率、高致残率及高死亡率的特点，被临床视为继肿瘤之后的难治疾病。其中70%为缺血性脑卒中，遗留明显的肢体功能障碍，给个人、家庭及社会带来沉重负担。针灸对缺血性脑卒中的临床疗效已被多次证实，其作用机制目前尚处于不断研究中。有研究发现利用针灸治疗缺血性脑卒中时，急性期、恢复期及后遗症期介入后疗效存在统计学差异，说明针灸治疗时机已然从恢复期/后遗症期逐渐转移至急性期[1]。近年来神经干细胞在修复受损神经方面的作用逐渐受到重视[2-3]，认为其自我更新及分化能力是治疗神经受损疾病的切入点，这亦予研究针刺的作用机制带来了新思路。……