Abstract
Background 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. Methods 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/β-catenin signaling pathway and p-GSK3β (ser9)/VEGF pathway in the injured brain tissues was assessed by western blotting and qRT-PCR. Results 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, β-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/β-catenin signaling pathway was activated after the treatment of scalp acupuncture. In contrast, dikkoppf-1 (DKK1) pretreatment, a specific inhibitor for the Wnt/β-catenin signaling pathway, inactivated the Wnt3a/β-catenin signaling pathway and exacerbated the infarct size induced by the cerebral IR injury on day 7. Conclusion Together, our findings demonstrated a mechanism whereby scalp acupuncture led to the activation of Wnt/β-catenin signaling pathway, promoting angiogenetic factor expression and restoring blood perfusion in the ischemic zone.
Highlights
Stroke is defined as the sudden death of brain cells due to vascular accidents caused by blockage or rupture of an artery to the brain
After routine skin preparation disinfection, the neck median was opened with a pair of scissors, and iris forceps was used to carefully dissect the common carotid artery (CCA) free from the vagus nerves without causing physical injury, and common carotid artery (CCA), internal carotid artery (ICA), and external carotid artery (ECA) were exposed. e trunk of the ICA was separated from the pterygopalatine artery (PPA), and an artery clamp was arranged at the starting position and the proximal part of the CCA. e ECA stump was gently pulled down, the 0.2 mm orifice was cut, and the tail end of the nylon line was gently pushed through the CCA bifurcation along the ICA into the skull to reach the middle cerebral artery (MCA)
Effect of Scalp Electroacupuncture on Cerebral Infarction Size and regional cerebral blood flow (rCBF) of the Cerebral Cortex after cerebral ischemia-reperfusion injury (CIRI). e Tetrazolium Chloride (TTC) staining and the rCBF of the cerebral cortex were used to measure the cerebral infarction area and the cerebral blood flow perfusion. e results of TTC staining (Figures 1(a) and 1(b)) showed that the volume of cerebral infarction induced by ischemia-reperfusion injury was significantly increased in the rats of the model group compared with that of the sham group (P < 0.01). e volume of cerebral infarction in the acupuncture group (AC) group was significantly lower than that in the model group (P < 0.05)
Summary
Stroke is defined as the sudden death of brain cells due to vascular accidents caused by blockage or rupture of an artery to the brain. E recent studies reveal that the Wnt/β-catenin signaling pathway plays important roles in promoting endothelial cell proliferation and maintaining blood-brain barrier homeostasis and has a synergistic effect with the GSK3β/VEGF pathway [15]. 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. Wnt3a, β-catenin, and cyclin D1 were 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/β-catenin signaling pathway was activated after the treatment of scalp acupuncture. Our findings demonstrated a mechanism whereby scalp acupuncture led to the activation of Wnt/β-catenin signaling pathway, promoting angiogenetic factor expression and restoring blood perfusion in the ischemic zone
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