Pu Yanpeng, Wang Zhen, Chu Haoran
Anhui University of Chinese Medicine Postdoctoral Research Station, Hefei 230000, China.
Department of Encephalopathy.
Nan Fang Yi Ke Da Xue Xue Bao. 2025 May 20;45(5):921-928. doi: 10.12122/j.issn.1673-4254.2025.05.04.
To evaluate the effect of eye acupuncture on neural function and angiogenesis of ischemic cerebral tissue in rats, and explore the roles of METTL3-mediated mA methylation and the HIF-1α/VEGF-A signal axis in mediating this effect.
Fifty SD rats were randomized into normal control group, sham-operated group, model group, eye acupuncture group and DMOG (a HIF-1α agonist) group. Rat models of cerebral ischemia/reperfusion injury (CIRI) were established using a modified thread thrombus method, and the changes in neurological deficits of the rats after interventions were evaluated. TTC and Nissl staining were used to examine the changes in infarction size and neuronal injury, and cerebral angiogenesis was detected by double-immunofluorescence staining. mA methylation modification level in the brain tissue was detected by ELISA, and RT-qPCR and Western blotting were used to detect the mRNA and protein expressions of METTL3 and HIF-1α/VEGF-A.
Compared with the control and sham-operated rats, the CIRI rats had significantly higher neurological deficit scores with larger cerebral infarction area, a greater number of CD31- and EDU-positive new vessels, higher expression levels of HIF-1α and VEGF-A, reduced number of Nissl bodies and m6A methylation level, and lowered METTL3 protein and mRNA expressions. All these changes were significantly improved by interventions with eye acupuncture after modeling or intraperitoneal injections of DMOG for 7 consecutive days prior to modeling, and the effects of the two interventions were similar.
Eye acupuncture can improve neurological deficits in CIRI rat models possibly by promoting cortical angiogenesis upregulating METTL3-mediated mA methylation and regulating the HIF-1α/VEGF-A signal axis.
评估眼针疗法对大鼠缺血脑组织神经功能及血管生成的影响，并探讨METTL3介导的m⁶A甲基化和HIF-1α/VEGF-A信号轴在介导该效应中的作用。
将50只SD大鼠随机分为正常对照组、假手术组、模型组、眼针组和DMOG（一种HIF-1α激动剂）组。采用改良线栓法建立大鼠脑缺血/再灌注损伤（CIRI）模型，评估干预后大鼠神经功能缺损的变化。采用TTC和尼氏染色检测梗死灶大小和神经元损伤的变化，通过双免疫荧光染色检测脑内血管生成。采用ELISA检测脑组织中m⁶A甲基化修饰水平，采用RT-qPCR和蛋白质免疫印迹法检测METTL3和HIF-1α/VEGF-A的mRNA和蛋白表达。
与对照组和假手术组大鼠相比，CIRI大鼠神经功能缺损评分显著更高，脑梗死面积更大，CD31和EDU阳性新血管数量更多，HIF-1α和VEGF-A表达水平更高，尼氏体数量减少，m⁶A甲基化水平降低，METTL3蛋白和mRNA表达降低。建模后采用眼针干预或在建模前连续7天腹腔注射DMOG均可显著改善上述所有变化，且两种干预效果相似。
眼针疗法可能通过促进皮质血管生成、上调METTL3介导的m⁶A甲基化并调节HIF-1α/VEGF-A信号轴来改善CIRI大鼠模型的神经功能缺损。
