百会透曲鬓针刺法对脑出血大鼠自噬 相关蛋白LC3A/B的影响研究

牛明明¹  邹伟^2△  于学平²

（1.黑龙江中医药大学临床医学院，黑龙江  哈尔滨，150040；2.黑龙江中医药大学附属第一医院，黑龙江  哈尔滨，150040）

中图分类号：R245.19      文献标识码：A    文章编号：1674-7860（2016）12-0015-04    证型：BDG

【摘  要】目的：观察“百会透曲鬓”针刺法对脑出血大鼠自噬相关蛋白LC3A/B表达的影响。方法：采用自体血注射的方法制作脑出血Wistar大鼠模型，将大鼠随机分为5组（空白对照组、模型对照组、抑制剂组、针刺组、抑制剂+针刺组），每组12只，再将各组随机分为4个时间亚组（造模后12 h、24 h、72 h、7 d），每亚组3只。空白对照组、模型对照组和抑制剂组分别在相应时间节点取材；抑制剂组于造模前15 min注射3-甲基腺嘌呤（3-MA）自噬抑制剂，针刺组和抑制剂+针刺组于造模成功后6 h开始以“百会透曲鬓” 针刺法针刺，1次/24 h，并于相应时间节点取材。分别于造模前和造模后6 h及处死前对大鼠神经功能进行评分，并用免疫组化的方法分析自噬相关蛋白LC3A/B的表达变化。结果：神经功能评分：造模前大鼠神经功能均为正常，造模后出现神经功能缺损症状，针刺组于处死前有改善且与干预时间成正相关，免疫组化结果显示：LC3A/B的病理变化表达在细胞质中，而在相同时间点，其程度高低依次为针刺组、模型组、抑制剂+针刺组，空白对照组和抑制剂组程度相似且明显低于前三组。结论：“百会透曲鬓”针刺法对脑出血后大鼠的神经功能缺损有改善作用且对自噬相关蛋白LC3A/B的表达有一定影响。

【关键词】自噬；针刺；脑出血；LC3A/B

【Abstract】Objective:To investigate the effect of Baihui-Qubin acupuncture on autophagy associatedBeclin-1 and LC3A/Bexpression in intracerebral hemorrhage (ICH) rat model. Methods: Intracerebralhemorrhage rat model was established by autologous blood injection. The Wistarrats were randomly divided into five groups: sham group, ICH model group,inhibitor group, acupuncture group and inhibitor + acupuncture group (12 rats ineach group); 4 sub-group was divided based on different time point (12hrs,24hrs, 72hrs, 7days), with 3 rats in each group. Brain tissues were harvestedat altered time point in sham group, ICH model group and inhibitor group. 3-MAinhibitor was injected at 15 minutes before autologous blood injection.Baihui-Qubin acupuncture was implemented in acupuncture group and inhibitor +acupuncture group 6hrs after ICH (once per 24hrs), brain tissues were alsoharvested at altered time point. Evaluate the neurological deficit 6hrs afterICH and before tissues being harvested. Immunohistochemical studies wereperformed to analyze the expression of Beclin-1 and LC3A/B in brain tissues. Results: Neurological deficitevaluation: the neurological behavior in acupuncture group was improvedassociated with the acupuncture involvement. Immunohistochemical results: LC3A/B pathology pattern was detected incytoplasm; the expression was increased in acupuncture group and reduced ininhibitor group as well as the inhibitor + acupuncture group which was slightlyhigher than that of the inhibitor group. Conclusion: Baihui-Qubin acupuncturemethod alleviated the neurological deficits and had effects on autophagyassociated Beclin-1 and LC3A/Bexpressions.

【Keywords】Autophagy;Acupuncture; Intracerebral hemorrhage (ICH); LC3A/B

doi:10.3969/j.issn.1674-7860.2016.12.008

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