目的探讨电针对成年弱视小鼠视皮层可塑性的调控作用及其机制。
方法采用随机数字表法将48只SPF级雄性健康3周龄昆明小鼠随机分为正常对照组、单眼形觉剥夺(MD)组和电针干预组，每组16只。除正常对照组外，其他小鼠均缝合右侧眼睑2周，建立成年MD弱视模型，电针干预组于小鼠5周龄时用电针刺激"太阳""睛明""风池"3个穴位4周。5周龄与9周龄时，分别检测各组小鼠主观视功能(探爪成功率)的变化。9周龄时，检测各组小鼠闪光视觉诱发电位(F-VEP)变化；采用Western blot法检测小鼠剥夺眼对侧视皮层中可塑性相关蛋白突触素(SYP)、突触蛋白1(SYN1)、轴索过度生长抑制因子A(Nogo-A)和轴索过度生长抑制因子受体(NgR)蛋白表达；采用免疫组织化学染色法检测小鼠视皮层中早期生长反应基因1(Egr-1)的表达。
结果9周龄时，正常对照组、MD组和电针干预组小鼠探爪成功率分别为(71.69±10.60)%、(25.54±10.09)%和(58.25±8.39)%，总体比较差异有统计学意义( F＝5.987， P＝0.006)，其中MD组小鼠探爪成功率明显低于正常对照组，电针干预组探爪成功率明显高于MD组，差异均有统计学意义(均 P<0.05)。各组小鼠F-VEP检查P2波振幅总体比较，差异有统计学意义( F＝63.710， P<0.001)，其中MD组小鼠剥夺眼P2波振幅较正常对照组和电针干预组明显降低，差异均有统计学意义(均 P<0.001)。各组小鼠剥夺眼对侧视皮层中SYP和SYN1蛋白相对表达量总体比较，差异均有统计学意义( F＝5.451、3.871，均 P<0.05)，其中MD组SYP和SYN1蛋白相对表达量明显低于正常对照组和电针干预组，差异均有统计学意义(均 P<0.05)。各组小鼠剥夺眼对侧视皮层中Nogo-A和NgR蛋白相对表达量总体比较，差异均有统计学意义( F＝4.188、3.942，均 P<0.05)，其中MD组Nogo-A和NgR蛋白相对表达量明显高于正常对照组和电针干预组，差异均有统计学意义(均 P<0.05)。免疫组织化学染色结果显示，与正常对照组比较，MD组小鼠剥夺眼对侧视皮层神经元Egr-1表达减少，棕褐色神经元突起难以分辨；与MD组比较，电针干预组小鼠剥夺眼对侧视皮层区域可见明显Egr-1阳性细胞表达，但表达强度弱于正常对照组。
结论电针刺激"太阳""睛明""风池"穴位可重新激活成年弱视小鼠视皮层可塑性，并改善弱视小鼠的视觉功能，其机制可能与调控Nogo-A/NgR信号通路有关。
ObjectiveTo investigate the regulatory effect of electroacupuncture on visual cortex plasticity in adult amblyopic mice and its mechanism.
MethodsForty-eight SPF male healthy 3-week-old Kunming mice were randomly divided into a normal control group, a monocular form deprivation (MD) group, and an electroacupuncture intervention group by the random number table method, with 16 mice in each group.Except for the normal control group, mice in the other groups had their right eyelids sutured for two weeks to establish an adult MD amblyopia model.The electroacupuncture intervention group received electroacupuncture stimulation at three acupoints, Taiyang (EX-HN5), Jingming (BL1), and Fengchi (GB20) for four weeks at five weeks of age.The subjective visual function (paw probing success rate) of each group of mice was measured at five and nine weeks of age, respectively.At nine weeks of age, the changes in flash-visual evoked potential (F-VEP) of mice in each group was detected.The expression of plasticity related proteins synaptophysin (SYP), synaptic protein 1 (SYN1), neurite overgrowth inhibitor A (Nogo-A), and Nogo receptor (NgR) proteins in the contralateral cortex of the deprived mouse eyes was detected by Western blot.Expression of early growth response gene 1 (Egr-1) in the visual cortex of mice was detected by immunohistochemical staining.This study was approved by the Animal Ethics Committee of Hunan Children's Hospital (No.HCHDWLL-2022-17), and the management and use of animals were in accordance with the Laboratory Animal Management and Use Guide of Hunan Children's Hospital.
ResultsAt nine weeks of age, the success rates of paw probing in the normal control group, MD group, and electroacupuncture intervention group were (71.69±10.60)%, (25.54±10.09)%, and (58.25±8.39)%, respectively, with a statistically significant overall difference ( F=5.987, P=0.006).Among them, the success rate of paw probing was significantly lower in the MD group than in the normal control group, and the electroacupuncture intervention group was significantly higher than in the MD group (both P<0.05).There was a significant overall difference in P2 wave amplitude in F-VEP examination among different groups of mice ( F=63.710, P<0.001), with lower P2 wave amplitude in the deprived eye of the MD group than in the normal control group and the electroacupuncture intervention group, and the differences were statistically significant (both P<0.001).There were significant differences in the expression levels of SYP and SYN1 proteins in the contralateral cortex of the deprived mouse eyes ( F=5.451, 3.871; both P<0.05).The relative expression levels of SYP and SYN1 proteins were significantly lower in the MD group than in the normal control group and electroacupuncture intervention group (all P<0.05).There were significant differences in levels of Nogo-A and NgR proteins in the contralateral cortex of the deprived mouse eyes ( F=4.188, 3.942, both P<0.05).The relative expression levels of Nogo-A and NgR proteins were significantly higher in the MD group than in the normal control group and the electroacupuncture intervention group (all P<0.05).The immunohistochemical staining results showed that compared with the normal control group, the MD group mice had a decrease in the expression of Egr-1 in the contralateral cortical neurons of the deprived eye, and the brown neuron protrusions were indistinguishable.Compared with the MD group, the electroacupuncture intervention group showed significant positive cell expression in the contralateral cortical area of the deprived eye, but the expression intensity was weaker than that of the normal control group.
ConclusionsElectroacupuncture treatment of Taiyang (EX-HN5), Jingming (BL1), and Fengchi (GB21) acupoints can reactivate the plasticity of the visual cortex in adult amblyopic mice and improve their visual function.The mechanism may be related to the regulation of Nogo-A/NgR signaling pathway.
李雯,付婧,王婵媛,等. 电针对成年弱视小鼠视皮层可塑性的调控作用及其机制[J]. 中华实验眼科杂志,2025,43(02)：106-113.
DOI:10.3760/cma.j.cn115989-20231227-00221版权归中华医学会所有。
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李雯：实施研究、分析数据、统计分析、起草文章；付婧：实施研究、采集数据、统计分析；王婵媛：设计实验、审阅修改文章；唐璟、刘玉婷：实施研究、采集数据；罗瑜琳：设计实验、指导研究、分析数据、对文章知识性内容作批评性审阅及定稿
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