Effects of acupuncture on the number of associated protein phosphorylation in brain tissues of MCAO rats based on protein microarray technique
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To investigate the effects of acupuncture on the number of associated phosphorylated proteins in brain tissues of middle cerebral artery occlusion (MCAO) rats, based on the protein microarray technique.
The MCAO model was prepared according to the modified occlusion method using occlusion lines. Forty healthy Sprague-Dawley (SD) rats were randomly divided into 4 groups using the lottery method: a sham operation group, a model group, a control point group and an acupoint group, with 10 rats in each group. Rats in the sham operation group and the model group only received binding without acupuncture. Rats in the acupoint group received acupuncture at Dazhui (GV 14), Baihui (GV 20) and Shuigou (GV 25); rats in the control point group received acupuncture at non-acupoint control points. The needle was twisted once for 1 min after insertion and another time in the middle of the 30 min needle retaining. Acupuncture was conducted once every 12 h for 6 consecutive times. At the end of the experiment, the neurological impairment score was collected, and cells of the ischemic brain tissues were extracted. The protein phosphorylation of the related signaling was detected using the 720 phosphorylated antibody microarray technique, and the differentially expressed proteins between groups were screened.
The neurological impairment scores after 72 h of treatment: compared with the sham operation group, the scores of the model group, the control point group and the acupoint group were significantly increased (P<0.01); compared with the model group, the scores of the acupoint group and the control point group were significantly decreased (P<0.01, P<0.05); the score of the acupoint group was better than that of the control point group (P<0.05). The results of the protein microarray: compared with the sham operation group, 48 proteins showed up-regulated phosphorylation (≥1.5 times) in the model group and the down-regulated was 28; compared with the model group, 35 proteins showed up-regulated phosphorylation in the control point group, and the down-regulated was 24. There were 29 proteins showing up-regulated phosphorylation in the acupoint group and the down-regulated was 51. The numbers of proteins involved in the function and signal transduction pathways were also different.
Acupuncture at Dazhui (GV 14), Baihui (GV 20) and Shuigou (GV 25) can effectively repair brain injury. The ischemic injury of brain tissue may be caused by imbalance of a variety of proteins, and acupuncture can promote brain tissue repair by multi-functional and multi-channel regulation of the protein disorders.
KeywordsAcupuncture Therapy Point Dazhui (GV 14) Point Baihui (GV 20) Point Shuigou (GV 26) Infarction Middle Cerebral Artery Protein Array Analysis Rats
参照线拴法并加以改良复制 MCAO 模型, 将40 只Sprague-Dawley (SD)健康大鼠按抽签法随机分为4 组, 即假手术组、模型组、对照点组和穴位组, 每组10 只。假手术组与模型组大鼠只捆绑不针刺, 穴位组大鼠针 刺大椎、百会、水沟, 对照点组大鼠针刺非穴对照点, 针后每穴捻转1 min, 中间捻转1 次, 留针30 min, 每12 h 针刺1 次, 连续6 次。实验结束后进行神经功能缺损评分, 提取缺血脑组织细胞, 采用720 磷酸化抗体蛋白芯片 技术观察各组大鼠脑组织损伤修复相关信号蛋白磷酸化的变化, 筛选出各组差异性表达的蛋白。
治疗72 h 后, 神经功能缺损评分: 与假手术组比较, 模型组、对照点组和穴位组评分显著增高(P<0.01); 与模型组比较, 穴<位组与对照点组评分降低(P<0.01, P<0.05); 穴位组评分优于对照点组(P<0.05)。蛋白芯片显示: 与假手术组比 较, 模型组磷酸化水平表达上调(≥1.5 倍)的蛋白数量为48 种, 下调的有28 种; 与模型组比较, 对照点组上调的 蛋白数量为35 种, 下调的有24 种; 穴位组上调蛋白数量为29 种, 下调的有51 种; 且在功能归属与信号转导途 径的归属上蛋白数量也不一致。
针刺大椎、百会和水沟能有效修复脑损伤; 脑组织的缺血性损伤可能由多种数量蛋白失调而联合致病, 而针刺则可以多功能、多途径地调整失调蛋白而促进脑组织修复。
关键词针刺疗法 穴 大椎 穴 百会 穴 水沟 梗死 大脑中动脉 蛋白质阵列分析 大鼠
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This work was supported by National Natural Science Foundation of China (国家自然科学基金项目, No. 81303051, No. 30901901); Province and Ministry Coconstruction Key Laboratory for Internal Medicine of Traditional Chinese Medicine of the Education Ministry of China (中医内科学省部共建教育部重点实验室, No. ZYNK201501); Tuina Department of Yueyang Hostpital Affiliated to Hunan University of Chinese Medicine State Clinical Key Specialty (湖南中医药大学附属岳阳医院推 拿科国家临床重点专科); Tuina Key Discipline of State Administration of Traditional Chinese Medicine (推拿学 国家中医药管理局重点学科).
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