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[电针预处理通过抑制大鼠大脑皮质组织自噬减轻脑缺血再灌注损伤]

[Electroacupuncture pretreatment alleviated cerebral ischemia-reperfusion injury via suppressing autophagy in cerebral cortex tissue in rats].

作者信息

Huang Ya-Guang, Yang Song-Bai, Du Li-Peng, Cai San-Jin, Feng Zhi-Tao, Mei Zhi-Gang

机构信息

Medical College of China Three Gorges University, Yichang 443002, Hubei Province, China; Affiliated Renhe Hospital of China Three Gorges University, Yichang 443000, Hubei Province.

Yichang Hospital of Traditional Chinese Medicine, Clinical College of Traditional Chinese Medicine of China Three Gorges University, Yichang 443003, Hubei Province.

出版信息

Zhen Ci Yan Jiu. 2019 Dec 25;44(12):867-72. doi: 10.13702/j.1000-0607.190307.

Abstract

OBJECTIVE

To investigate the effect of electroacupuncture (EA) preconditioning on autophagy in cerebral cortex tissues of rats with cerebral ischemia-reperfusion injury (CIRI), so as to explore its mechanisms underlying improvement of CIRI.

METHODS

Thirty-three male Sprague-Dawley rats were randomly divided into sham operation, model and EA groups (=11 in each group). EA (2 Hz/15 Hz, 1 mA) was applied to "Baihui"(GV20), "Quchi" (LI11) and "Zusanli" (ST36) for 30 min, once daily for 5 days, followed by establishment of CIRI model by occlusion of the middle cerebral artery (MCAO) for 1.5 h and reperfusion for 24 h. The neurological deficit score was assessed in reference to Longa's methods, and the infarct volume assessed by 2,3,5-triphenyltetrazolium chloride staining. The density of dendrite spines of neurons in the ischemic cerebral cortex tissue was detected by Golgi's staining, the autophagosome observed by electron microscopy, and the expression levels of microtubule-associated protein 1 light chain 3 (LC3) and p62 (a selective autophagy substrate) were detected by Western blot.

RESULTS

Compared with the sham operation group, the neurological deficit score and infarct volume were significantly increased (<0.01), the number of autophagosomes and the ratio of LC3-Ⅱ/LC3-Ⅰ also significantly increased (<0.01), while the expression level of p62 was notably decreased in the model group (<0.01). Following the intervention and in comparison with the model group, the neurological deficit score and infarct volume were significantly reduced (<0.01), the number of autophagosomes and the ratio of LC3-Ⅱ/LC3-Ⅰ obviously decreased (<0.01), and the expression of p62 was significantly up-regulated in the EA group (<0.01).

CONCLUSION

EA pretreatment is effective in improving CIRI in rats, which may be realized through suppressing autophagy in the ischemic cerebral cortex tissue.

摘要

目的

探讨电针预处理对脑缺血再灌注损伤(CIRI)大鼠大脑皮质组织自噬的影响,以探究其改善CIRI的机制。

方法

将33只雄性Sprague-Dawley大鼠随机分为假手术组、模型组和电针组(每组11只)。采用2 Hz/15 Hz、1 mA的电针刺激“百会”(GV20)、“曲池”(LI11)和“足三里”(ST36),每次30分钟,每日1次,连续5天,随后通过大脑中动脉闭塞(MCAO)1.5小时并再灌注24小时建立CIRI模型。参照Longa法评估神经功能缺损评分,通过2,3,5-三苯基氯化四氮唑染色评估梗死体积。采用高尔基染色检测缺血大脑皮质组织中神经元树突棘的密度,通过电子显微镜观察自噬体,采用蛋白质免疫印迹法检测微管相关蛋白1轻链3(LC3)和p62(一种选择性自噬底物)的表达水平。

结果

与假手术组相比,模型组神经功能缺损评分和梗死体积显著增加(<0.01),自噬体数量和LC3-Ⅱ/LC3-Ⅰ比值也显著增加(<0.01),而p62表达水平显著降低(<0.01)。干预后,与模型组相比,电针组神经功能缺损评分和梗死体积显著降低(<0.01),自噬体数量和LC3-Ⅱ/LC3-Ⅰ比值明显降低(<0.01),p62表达显著上调(<0.01)。

结论

电针预处理可有效改善大鼠CIRI,其机制可能是通过抑制缺血大脑皮质组织中的自噬实现的。

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