[Effect of ZHU Lian's acupuncture stimulation method on neurological function in rats with ischemic stroke based on...

To observe the effects of the ZHU Lian's acupuncture stimulation method on neurological function and miR-34a-5p/tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein γ (YWHAG) pathway in rats with ischemic stroke (IS), and to explore its mechanism in alleviating neurological impairment in IS.

A total of 72 clean-grade male SD rats, with 10 rats were randomly selected as the sham operation group, and the remaining rats were subjected to suture occlusion method to establish an IS model. Fifty successfully modeled rats were randomly divided into a model group, an acupuncture group, a positive drug group, an acupuncture+agonist control group, and an acupuncture+agonist group, with 10 rats in each group. The rats in the acupuncture group were treated with ZHU Lian's acupuncture stimulation method at "Shuigou" (GV26), "Baihui" (GV20), "Dazhui" (GV14), and bilateral "Neiguan" (PC6) and "Hegu" (LI4). The rats in the positive drug group were treated with 10.8 mg/kg nimodipine via gavage. The rats in the acupuncture+agonist group and acupuncture+agonist control group were treated with intracerebroventricular injections of miR-34a-5p agonist (miR-34a-5p agomir) and negative control (agomir NC), respectively, at a dose of 5μL, followed by the same acupuncture protocol. All treatments were performed once daily for 5 days, followed by 2 days off, continuing for 21 days. After intervention, neurological deficits were evaluated using the Zea-Longa score. The infarct volume percentage was measured by TTC staining. Neuronal damage and Nissl bodies in the ischemic cortex were observed by Nissl staining. Neuronal apoptosis rate in the ischemic cortex was assessed by TUNEL staining. Relative expression levels of miR-34a-5p and YWHAG mRNA in the ischemic cortex were measured by RT-qPCR. Protein levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and YWHAG were detected by Western blot. A dual-luciferase assay was used to verify the relationship between miR-34a-5p and YWHAG.

Compared with the sham operation group, the Zea-Longa scores, the percentage of cerebral infarct volume, the neuronal apoptosis rate in the ischemic cortical tissue, the expression levels of IL-6, TNF-α, and miR-34a-5p mRNA in the model group were increased (P<0.05), and the expression of YWHAG mRNA and protein was decreased (P<0.05). Compared with the model group, the Zea-Longa scores, the percentage of cerebral infarct volume, the neuronal apoptosis rate in the ischemic cortical tissue, the expression levels of IL-6, TNF-α, and miR-34a-5p mRNA in the acupuncture group, the positive drug group, and the acupuncture+agonist control group were decreased (P<0.05), and the expression of YWHAG mRNA and protein was increased (P<0.05). Compared with the acupuncture+agonist control group, the Zea-Longa scores, the percentage of cerebral infarct volume, the neuronal apoptosis rate in the ischemic cortical tissue, the expression levels of IL-6, TNF-α, and miR-34a-5p mRNA in the acupuncture+ agonist group were increased (P<0.05), and the expression of YWHAG mRNA and protein was decreased (P<0.05). In addition, the miR-34a-5p and YWHAG have a targeted regulatory relationship (P<0.05).

The ZHU Lian 's acupuncture stimulation method could improve neurological function in IS model rats, possibly by modulating the miR-34a-5p/YWHAG pathway.