基于神经细胞凋亡信号通路探讨针刺延长脑梗死的溶栓时间窗效应  

Effect of acupuncture on neurological function, cerebral infarction volume, thrombolysis time window and cerebral cell apoptosis signaling pathway in cerebral infarction rats

作　　者：顾亚会 张新昌 徐文韬 张奥 张智慧 姜思媛 常思琦 倪光夏 GU Ya-hui;ZHANG Xin-chang;XU Wen-tao;ZHANG Ao;ZHANG Zhi-hui;JIANG Si-yuan;CHANG Si-qi;NI Guang-xia(College of Acupuncture-moxibustion and Tuina,Nanjing University of Chinese Medicine,Nanjing 210046,China)

机构地区：[1]南京中医药大学针灸推拿学院,南京210046

出　　处：《针刺研究》2020年第3期209-214,226,共7页Acupuncture Research

基　　金：国家自然科学基金项目(No.81674067);江苏省高校自然科学研究项目(No.18KJB360007)。

摘　　要：目的:观察针刺对脑梗死急性期大鼠大脑皮层梗死体积及相关蛋白的影响,探讨针刺延长脑梗死的溶栓时间窗效应及其可能的机制。方法:将48只大鼠随机分为假手术组、模型组、溶栓4.5 h组、溶栓6 h组、溶栓9 h组、针刺+溶栓4.5 h组、针刺+溶栓6 h组、针刺+溶栓9 h组,每组6只。以改良自体血栓栓塞法建立大鼠脑梗死模型。各溶栓组仅在相应时间通过尾静脉注射重组人组织型纤溶酶原激活剂(rt-PA,10 mg/kg)进行溶栓;造模成功后2 h,各针刺溶栓组给予针刺"水沟"及双侧"内关",留针30 min,并在相应时间进行和各溶栓组相同的干预处理;假手术组和模型组仅给予相同的固定。分别对各组大鼠在造模成功后2 h和24 h进行神经行为学评分。各组均在造模后24 h神经行为学评分完成后处死大鼠并取材,用2,3,5-三苯基氯化四氮唑(TTC)染色法比较各组大鼠脑梗死体积百分比,Western blot法检测各组大鼠大脑皮层多聚ADP核糖聚合酶-1(PARP1)、凋亡诱导因子(AIF)和核酸内切酶G(Endo-G)的表达。结果:与假手术组比较,模型组大鼠神经行为学评分、脑梗死体积百分比、PARP1、AIF和Endo-G蛋白表达水平均明显升高(P<0.01,P<0.05)。与模型组比较,溶栓4.5 h组、针刺+溶栓4.5 h组、针刺+溶栓6 h组神经行为学评分、脑梗死体积百分比、AIF和Endo-G表达水平均明显降低(P<0.05),溶栓4.5 h组、针刺+溶栓4.5 h组的PARP1蛋白表达明显降低(P<0.05)。与溶栓4.5 h组比较,针刺+溶栓4.5 h组的神经行为学评分、脑梗死体积百分比及AIF、PARP1、Endo-G蛋白表达均明显降低(P<0.05)。与溶栓6 h组比较,针刺+溶栓6 h组神经行为学评分、脑梗死体积百分比及AIF蛋白表达均明显降低(P<0.05)。与溶栓9 h组比较,针刺+溶栓9 h组Endo-G的表达显著下调(P<0.05)。结论:针刺+溶栓干预可减少脑梗死体积,改善溶栓后大鼠神经功能损伤,延长溶栓时间窗。针刺延长脑Objective To observe the effect of acupuncture(Acupunct) on cerebral infarction volume and expression of poly ADP ribose polymerase 1(PARP1), apoptosis-inducing factor(AIF) and endonuclease G(Endo-G) in the cerebral cortex tissue at different time-points after cerebral ischemia(CI) in acute cerebral infarction rats, so as to explore its underlying mechanisms in prolonging time window of thrombolysis. Methods Forty-eight SD rats were randomly divided into sham operation, model, intravenous thrombolysis(IVT)-4.5 h, IVT-6 h, IVT-9 h, Acupunct+IVT-4.5 h, Acupunct +IVT-6 h, Acupunct+IVT-9 h groups(n=6 in each group). The CI model was established by using modified autologous thromboembolism via the right common carotid artery. Two hours after modeling, rats of the Acupunct groups received Acupunct stimulation of "Shuigou"(GV26) and bilateral "Neiguan"(PC6) for 30 min. Thrombolysis was conducted by injection of recombinant human tissue-type plasminogen activator(rt-PA, 10 mg/kg) via caudal vein. The neurological deficit was assessed with reference to Bederson’s methods. 2,3,5-triphenyltetrazolium chloride(TTC) staining was used to assess the cerebral infarction volume, and the expression of cerebral PARP1, AIF and Endo-G proteins detected by Western blot. Results Compared with the sham operation group, the neurological score and percentage of cerebral infarction volume, expression levels of PARP1, AIF and Endo-G proteins were significantly increased in the model group(P<0.01,P<0.05). After the intervention, modeling-induced increase of the aforementioned indexes was reversed in the IVT-4.5 h, Acupunct+IVT-4.5 h and Acupunct+IVT-6 h groups(P<0.05), except PARP1 expression of the Acupunct+IVT 6 h group(P>0.05). The levels of neurological score, percentage of cerebral infarction volume, and AIF expression were significantly lower in both the Acupunct+IVT 4.5 h and Acupunct+IVT-6 h groups than in the simple IVT-4.5 h and simple IVT-6 h groups, respectively(P<0.05), and the expression levels of PARP1 and Endo-G proteins we

关 键 词：溶栓时间窗 脑梗死 针刺 多聚ADP核糖聚合酶-1 凋亡诱导因子 核酸内切酶G 

分 类 号：R245.31[医药卫生—针灸推拿学]