Zhang Yi-Min, Chen Sheng-Xin, Dai Qiu-Fu, Jiang Shu-Ting, Chen Ai-Lian, Tang Chun-Zhi, Zhang Yu-Qing
Department of Traditional Chinese Medicine, School of Medicine, Jinan University, 510632, Guangzhou, China.
Clinical Medical School of Acupuncture Rehabilitation, Guangzhou University of Traditional Chinese Medicine, 510405, Guangzhou, China.
Chin J Integr Med. 2018 Jul;24(7):537-544. doi: 10.1007/s11655-015-1969-9. Epub 2015 Apr 24.
To observe the effect of acupuncture on the Notch signaling pathway in rats with traumatic brain injury and to explore the pathogenesis of acupuncture intervention on traumatic brain injury.
Feeney's freefall epidural impact method was used to establish a traumatic brain injury model in rats; the rats were randomly divided into a normal group, sham operation group, model group and acupuncture group. Acupuncture was performed in the Baihui (DU 20), Shuigou (DU 26), Fengfu (DU 16), Yamen (DU 15) and Hegu (LI 4) acupoints in the rat, and Yamen was punctured via Fengfu. Then, the rats in each group were randomly divided into three subgroups, namely the day 3 subgroup, day 7 subgroup and day 14 subgroup according to treatment duration. The modified neurological severity scores (mNss) method was used to perform neurobehavioral scoring for evaluating the degree of injury in the rats. The hematoxylin-eosin (HE) staining method was used to observe the pathological change in the brain tissue of rats in each group. Real-time fluorescent quantitative polymerase chain reaction (Q-PCR) technology was used to detect changes in the Notch1, Hes1 and Hes5 gene expression levels in the cortex on the injured side. Western blot was used to detect the protein expression changes.
One day after modeling, the mNss scores in the model group and in the acupuncture group were significantly higher than those in the normal and sham operation groups (P<0.01) ; there was no statistically significant difference between the normal group and the sham operation group. The scores decreased with increased treatment time, and the scores in the acupuncture group decreased more significantly than those in the model group (P<0.01). The pathological examination by the HE staining method demonstrated that the brain tissue of the rats in the acupuncture and model groups relatively significantly changed. The Notch1 gene expression level in the acupuncture group was significantly higher than the level in all of the other groups (P<0.01) ; the Hes1 and Hes5 gene expression levels were also higher in the acupuncture group. The expression changes of the Notch1 and Hes1 protein were consistent with that of mRNA. In each experimental group, the mNss score and the pathological results by the HE staining method were consistent with the mRNA results.
Acupuncture could significantly promote high expression levels of Notch1, Hes1 and Hes5 in the brain tissue of traumatic brain injury rats. Therefore, acupuncture might be an important intervention for inducing endogenous stem cell proliferation and for promoting nerve repair.
观察针刺对创伤性脑损伤大鼠Notch信号通路的影响，探讨针刺干预创伤性脑损伤的发病机制。
采用Feeney自由落体硬脑膜冲击法建立大鼠创伤性脑损伤模型；将大鼠随机分为正常组、假手术组、模型组和针刺组。对大鼠百会（督脉20）、水沟（督脉26）、风府（督脉16）、哑门（督脉15）和合谷（手阳明大肠经4）穴位进行针刺，经风府针刺哑门。然后，根据治疗时间将每组大鼠随机分为三个亚组，即第3天亚组、第7天亚组和第14天亚组。采用改良神经功能缺损评分（mNss）法对大鼠进行神经行为评分，以评估大鼠损伤程度。采用苏木精-伊红（HE）染色法观察各组大鼠脑组织的病理变化。采用实时荧光定量聚合酶链反应（Q-PCR）技术检测损伤侧皮质Notch1、Hes1和Hes5基因表达水平的变化。采用蛋白质免疫印迹法检测蛋白表达变化。
造模后1天，模型组和针刺组的mNss评分显著高于正常组和假手术组（P<0.01）；正常组和假手术组之间无统计学差异。评分随治疗时间延长而降低，针刺组评分降低比模型组更显著（P<0.01）。HE染色法病理检查显示，针刺组和模型组大鼠脑组织变化较明显。针刺组Notch1基因表达水平显著高于其他所有组（P<0.01）；针刺组Hes1和Hes5基因表达水平也较高。Notch1和Hes1蛋白的表达变化与mRNA一致。各实验组中，mNss评分和HE染色法病理结果与mRNA结果一致。
针刺可显著促进创伤性脑损伤大鼠脑组织中Notch1、Hes1和Hes5的高表达。因此，针刺可能是诱导内源性干细胞增殖和促进神经修复的重要干预措施。
