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针刺对脑梗死大鼠神经功能、脑梗死体积、溶栓时间窗及脑细胞凋亡信号通路的影响

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

作者信息

Gu Ya Hui, Zhang Xin Chang, Xu Wen Tao, Zhang Ao, Zhang Zhi Hui, Jiang Si Yuan, Chang Si Qi, Ni Guang Xia

机构信息

College of Acupuncture-moxibustion and Tuina, Nanjing University of Chinese Medicine, Nanjing 210046, China.

出版信息

Zhen Ci Yan Jiu. 2020 Mar 25;45(3):209-14. doi: 10.13702/j.1000-0607.190635.

Abstract

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 (=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 (<0.01,<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 (<0.05), except PARP1 expression of the Acupunct+IVT 6 h group (>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 (<0.05), and the expression levels of PARP1 and Endo-G proteins were obviously lower in the Acupunct+IVT-4.5 h group than in the IVT-4.5 h group (<0.05). Endo-G proteins were obviously lower in the Acupunct+IVT-9 h group than in the IVT-9 h group (<0.05).

CONCLUSION

Acupuncture may improve neurological function, reduce cerebral infarction volume and prolong the time window of thrombolysis in CI rats, which may be associated with its effect in suppressing AIF/PARP1/ Endo-G signaling.

摘要

目的

观察针刺对急性脑梗死大鼠脑缺血(CI)后不同时间点脑梗死体积及大脑皮质组织中多聚ADP核糖聚合酶1(PARP1)、凋亡诱导因子(AIF)和核酸内切酶G(Endo-G)表达的影响,以探讨其延长溶栓时间窗的潜在机制。

方法

将48只SD大鼠随机分为假手术组、模型组、静脉溶栓(IVT)-4.5 h组、IVT-6 h组、IVT-9 h组、针刺+IVT-4.5 h组、针刺+IVT-6 h组、针刺+IVT-9 h组(每组n=6)。采用改良自体血栓栓塞法经右侧颈总动脉建立CI模型。造模后2 h,针刺组大鼠接受“水沟”(GV26)和双侧“内关”(PC6)针刺刺激30 min。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。参照Bederson法评估神经功能缺损。采用2,3,5-氯化三苯基四氮唑(TTC)染色评估脑梗死体积,采用蛋白质免疫印迹法检测大脑PARP1、AIF和Endo-G蛋白的表达。

结果

与假手术组比较,模型组神经功能评分、脑梗死体积百分比、PARP1、AIF和Endo-G蛋白表达水平均显著升高(P<0.01,P<0.05)。干预后,IVT-4.5 h组、针刺+IVT-4.5 h组和针刺+IVT-6 h组上述指标较造模后升高情况得到改善(P<0.05),针刺+IVT 6 h组PARP1表达除外(P>0.05)。针刺+IVT 4.5 h组和针刺+IVT-6 h组神经功能评分、脑梗死体积百分比及AIF表达水平分别显著低于单纯IVT-4.5 h组和单纯IVT-6 h组(P<0.05),针刺+IVT-4.5 h组PARP1和Endo-G蛋白表达水平显著低于IVT-4.5 h组(P<0.05)。针刺+IVT-9 h组Endo-G蛋白表达水平显著低于IVT-9 h组(P<0.05)。

结论

针刺可改善CI大鼠神经功能,减小脑梗死体积,延长溶栓时间窗,其机制可能与抑制AIF/PARP1/Endo-G信号通路有关。

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