Objective: To observe the effect of electroacupuncture (EA) on modified neurological severity score (mNSS), cerebral infarction volume, expression of Lim domain kinase-1 (LIMK1) and slingshot homolog-1 (SSH1) proteins, Cofilin rod formation and neural cell apoptosis in rats with ischemic stroke (IS), so as to explore its mechanisms underlying improvement of IS.
Methods: Male SD rats were randomly divided into normal, model and EA groups, with 13 rats in each group. The IS model was established by occlusion of the middle cerebral artery (MCAO) according to Zea Longa's method. EA was applied to "Quchi" (LI11) and "Zusanli" (ST36) for 30 min, once a day for 7 consecutive days. The behavioral changes of mNSS were observed before and after modeling. The volume of cerebral infarction was measured by using a small animal magnetic resonance imaging. The protein expressions of LIMK1 and SSH1 in the cerebral ischemic tissues were detected by Western blot. The density of Cofilin rod and neural cell apoptosis in cerebral ischemic area were determined by immunofluorescence staining and TUNEL staining, separately.
Results: After modeling, the mNSS score, cerebral infarction volume ratio, expression level of SSH1, density of Cofilin rod and the number of apoptotic cells were significantly increased (P<0.01), while the expression level of LIMK1 protein was obviously decreased in the model group relevant to the normal group (P<0.01). After 7 days' treatment, all the increased and decreased levels of the indexes mentioned above were reversed in the EA group relevant to the model group (P<0.01, P<0.05).
Conclusion: EA of LI11 and ST36 can improve neurological function and reduce infarction range in MCAO rats, which may be related to its action in regulating the expression of LIMK1 and SSH1, inhibiting the formation of Cofilin rod and reducing apoptosis of neural cells.
目的：观察电针对缺血性脑卒中大鼠改良神经功能缺损量表(mNSS)评分、脑梗死体积、缺血半暗带区脑组织中单丝氨酸蛋白激酶1(LIMK1)及丫叉同源物1(SSH1)蛋白表达、丝切蛋白棒状小体(Cofilin rod)密度及神经细胞凋亡的影响, 探讨电针治疗缺血性脑卒中的可能机制。方法：SD大鼠随机分为正常组、模型组及电针组, 每组13只。采用Zea Longa线栓法制备大脑中动脉栓塞(MCAO)大鼠模型。给予电针组大鼠右侧“曲池”“足三里”电针治疗,30 min/次, 1次/d, 连续7 d。观察各组大鼠mNSS评分；用小动物磁共振成像系统检测大鼠脑梗死体积；Western blot法检测缺血半暗带区脑组织中LIMK1、SSH1蛋白表达量；免疫荧光染色法检测缺血半暗带区Cofilin rod密度；TUNEL法检测神经细胞凋亡数。结果：与正常组比较, 模型组大鼠mNSS评分、脑梗死体积百分比、缺血半暗带区脑组织中SSH1蛋白表达、Cofilin rod密度、凋亡细胞数量均上升(P<0.01), LIMK1蛋白表达明显降低(P<0.01)；治疗7 d后, 与模型组比较, 电针组mNSS评分、脑梗死体积百分比及缺血半暗带区脑组织中SSH1蛋白表达、Cofilin rod密度、凋亡细胞数量均下降(P<0.01, P<0.05), LIMK1表达升高(P<0.05)。结论：电针“曲池”“足三里”对MCAO大鼠具有神经修复作用, 其机制可能与调控LIMK1、SSH1蛋白表达, 抑制Cofilin rod形成, 减少神经细胞凋亡有关。.
Keywords: Apoptosis; Cofilin rod; Electroacupuncture; Ischemic stroke; Neurological function.