Shi Shuai, Wang Mingye, Liu Xiaofang, Han Shengwang, Zhu Pengyu
The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine, No. 411, The Street of Guogeli, Harbin 150001, Heilongjiang, China.
College of Integrated Traditional Chinese and Western Medicine, Hebei Medical University of Chinese Medicine, No. 326, The South of Xinshi Street, Shijiazhuang 050091, Hebei, China.
Evid Based Complement Alternat Med. 2022 Mar 14;2022:1649605. doi: 10.1155/2022/1649605. eCollection 2022.
Scalp acupuncture is a contemporary acupuncture method based on the fundamental theories of traditional acupuncture, which has been widely used in patients with stroke in China. However, the effectiveness is controversial due to lack of solid experimental evidence.
In this study, a rat model of cerebral ischemia-reperfusion injury (CIRI) was established by the middle cerebral artery occlusion/recirculation. The efficacy of scalp acupuncture against CIRI was evaluated by the mNSS scores, TTC staining for brain slices, and laser Doppler perfusion imaging. Immunohistochemical staining for angiogenetic factors indicated the vascularization after CIRI, including VEGF, Ang2, and bFGF. Activation of the Wnt/-catenin signaling pathway and p-GSK3 (ser9)/VEGF pathway in the injured brain tissues was assessed by western blotting and qRT-PCR.
On the 7, 14, and 21 days after CIRI, scalp acupuncture could reduce the mNSS scores, decrease the cerebral infarction area, and accelerate the recirculation of ischemic brain tissues. VEGF, FLK1, bFGF, and Ang2 were upregulated on both the mRNA and protein levels in the ischemic brain tissues of the AC group, suggesting that the recirculation might result from angiogenesis, which was also confirmed with the IHC staining in the angiogenetic markers of VEGF, Ang2, and bFGF. Moreover, Wnt3a, -catenin, and cyclin D1 were also upregulated on both the mRNA and protein levels in the ischemic brain tissues of the AC group on day 7, 14, and 21, indicating that the Wnt/-catenin signaling pathway was activated after the treatment of scalp acupuncture. In contrast, dikkoppf-1 (DKK1) pretreatment, a specific inhibitor for the Wnt/-catenin signaling pathway, inactivated the Wnt3a/-catenin signaling pathway and exacerbated the infarct size induced by the cerebral IR injury on day 7.
Together, our findings demonstrated a mechanism whereby scalp acupuncture led to the activation of Wnt/-catenin signaling pathway, promoting angiogenetic factor expression and restoring blood perfusion in the ischemic zone.
头皮针是基于传统针灸基本理论的一种现代针灸方法，在中国已广泛应用于中风患者。然而，由于缺乏确凿的实验证据，其有效性存在争议。
在本研究中，通过大脑中动脉闭塞/再灌注建立大鼠脑缺血再灌注损伤（CIRI）模型。通过改良神经功能缺损评分（mNSS）、脑片TTC染色和激光多普勒灌注成像评估头皮针对CIRI的疗效。血管生成因子的免疫组织化学染色显示CIRI后的血管生成情况，包括血管内皮生长因子（VEGF）、血管生成素2（Ang2）和碱性成纤维细胞生长因子（bFGF）。通过蛋白质印迹法和定量逆转录聚合酶链反应（qRT-PCR）评估受损脑组织中Wnt/β-连环蛋白信号通路和p-糖原合成酶激酶3（ser9）/VEGF通路的激活情况。
在CIRI后的第7、14和21天，头皮针可降低mNSS评分，减小脑梗死面积，并加速缺血脑组织的再灌注。针刺组缺血脑组织中VEGF、血管内皮生长因子受体1（FLK1）、bFGF和Ang2在mRNA和蛋白水平均上调，提示再灌注可能是由血管生成所致，这也通过VEGF、Ang2和bFGF血管生成标志物的免疫组织化学染色得到证实。此外，在第7、14和21天，针刺组缺血脑组织中Wnt3a、β-连环蛋白和细胞周期蛋白D1在mRNA和蛋白水平也上调，表明头皮针治疗后Wnt/β-连环蛋白信号通路被激活。相反，Wnt/β-连环蛋白信号通路的特异性抑制剂迪克kopf-1（DKK1）预处理可使Wnt3a/β-连环蛋白信号通路失活，并在第7天加重脑缺血再灌注损伤诱导的梗死面积。
总之，我们的研究结果证明了一种机制，即头皮针可导致Wnt/β-连环蛋白信号通路激活，促进血管生成因子表达并恢复缺血区的血液灌注。
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