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眼针疗法对脑缺血再灌注损伤大鼠脑组织自噬的影响

[Effect of eye acupuncture on autophagy in brain tissue of rats with cerebral ischemia-reperfusion injury].

作者信息

Pu Yan-Peng, Wang Peng-Qin

机构信息

Liaoning University of Traditional Chinese Medicine, Shenyang 110032, China.

Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang 110032.

出版信息

Zhen Ci Yan Jiu. 2021 Feb 25;46(2):100-5. doi: 10.13702/j.1000-0607.200210.

Abstract

OBJECTIVE

to observe the effect of eye acupuncture on cerebral blood flow and autophagy of cerebral tissue in rats with cerebral ischemia-reperfusion injury, so as to explore the mechanism of eye acupuncture underlying improvement of cerebral ischemia-reperfusion injury.

METHODS

Fifty SD rats were randomly divided into sham operation, model, eye acupuncture, inhibitor and enhancer groups,with 10 rats in each group. The rat model of cerebral ischemia-reperfusion injury was established by occlusion of the middle cerebral artery for 1.5 h. The rats in the eye acupuncture group were treated with eye acupuncture for 30 minutes immediately, 12 h and 24 h after the modeling. Rats in the inhibitor group and enhancer group were given intracerebroventricular injection of autophagy agonist 3-Methyladenine or autophagy inducer Rapamycin 30 min before modeling. Longa's scoring method was used to evaluate the neurological function. The blood flow velocity of the cerebral cortex was mea-sured with a laser doppler blood flow meter, and the neuron damage in the brain tissue was observed with Nissl staining. The expressions of autophagy-related protein Beclin-1, microtubule-associated protein 1 light chain 3 (LC3), and p62 were measured by Western blot.

RESULTS

Compared with the sham operation group, the neurological score of the model group was significantly increased (P<0.01); the blood flow volume and blood flow speed were significantly reduced (P<0.01); the number of Nissl bodies in the ischemic brain tissue decreased (P<0.01); Beclin-1 protein expression level and LC3-II/LC3-I increased, while p62 expression level decreased(P<0.01). After intervention and in comparison with the model group, the neurological scores in the eye acupuncture group and inhibitor group decreased (P<0.01); blood flow volumn and blood flow speed significantly increased (P<0.01); the number of Nissl bodies increased (P<0.01); the expression level of Beclin-1 protein and LC3-II/LC3-I decreased, and the expression level of p62 increased (P<0.01). There was no significant difference between the enhancer and the model groups in the abovementioned indexes(P>0.05).

CONCLUSION

Eye acupuncture can improve the neurological function of rats with cerebral ischemia-reperfusion injury, which may be related to accelerating cerebral blood flow and inhibiting autophagy in the ischemic brain.

摘要

目的

观察眼针疗法对脑缺血再灌注损伤大鼠脑血流量及脑组织自噬的影响,以探讨眼针改善脑缺血再灌注损伤的作用机制。

方法

将50只SD大鼠随机分为假手术组、模型组、眼针组、抑制剂组和增强剂组,每组10只。采用大脑中动脉闭塞1.5小时的方法建立脑缺血再灌注损伤大鼠模型。眼针组大鼠于造模后即刻、12小时和24小时给予眼针治疗30分钟。抑制剂组和增强剂组大鼠在造模前30分钟经脑室注射自噬激动剂3-甲基腺嘌呤或自噬诱导剂雷帕霉素。采用Longa评分法评估神经功能。用激光多普勒血流仪测量大脑皮质血流速度,用尼氏染色观察脑组织神经元损伤情况。采用蛋白质免疫印迹法检测自噬相关蛋白Beclin-1、微管相关蛋白1轻链3(LC3)及p62的表达。

结果

与假手术组比较,模型组神经功能评分显著升高(P<0.01);脑血流量和血流速度显著降低(P<0.01);缺血脑组织尼氏体数量减少(P<0.01);Beclin-1蛋白表达水平及LC3-II/LC3-I升高,而p62表达水平降低(P<0.01)。干预后,与模型组比较,眼针组和抑制剂组神经功能评分降低(P<0.01);脑血流量和血流速度显著升高(P<0.01);尼氏体数量增多(P<0.01);Beclin-1蛋白表达水平及LC3-II/LC3-I降低,p62表达水平升高(P<0.01)。增强剂组与模型组上述指标比较差异无统计学意义(P>0.05)。

结论

眼针可改善脑缺血再灌注损伤大鼠的神经功能,其机制可能与加快脑血流及抑制缺血脑组织自噬有关。

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