Yang Qi-Qi, Yang Jun, Ye Min, Wang Ke-Po, Zhou Xin-Hua, Li Wei-Ran, Li Fei
Rehabilitation Department, The Second Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230061, China.
Acupuncture and Rehabilitation Department, The First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230031.
Zhen Ci Yan Jiu. 2025 Mar 25;50(3):260-269. doi: 10.13702/j.1000-0607.20240550.
OBJECTIVES: To investigate the effect of moxibustion on hippocampal mitochondrial dynamics-related proteins in rats with vascular dementia (VD), so as to reveal the underlying mechanisms of moxibustion in treating VD. METHODS: SD rats were randomly divided into sham operation, model, moxibustion and medication groups, with 12 rats in each group. The VD model was prepared using an improved bilateral common carotid artery ligation method. Mild moxibstion was applied to "Shenting" (GV24), "Baihui" (GV20) and "Dazhui" (GV14) for 20 min, once daily for 6 days per week. Rats of the medication group were treated with oral administration of nimodipine (12 mg/kg) once daily. All above interventions were performed for 4 weeks. Learning and memory abilities were assessed using Morris water maze test. Histopathological changes of hippocampus was observed with HE staining. Mitochondrial membrane potential (Δψm) and reactive oxygen species (ROS) levels in hippocampal tissue were measured by flow cytometry. The mRNA and protein expression levels of optic atrophy protein 1 (Opa1), mitochondrial fusion protein 1 (Mfn1), mitochondrial fusion protein 2 (Mfn2), dynamics-related protein 1 (Drp1), fission protein 1 (Fis1), mitochondrial fission factor (Mff) in hippocampus was detected by PCR or Western blot, respectively. RESULTS: Compared with the sham operation group, the escape latency was prolonged (<0.01), the times of crossing the original platform were reduced (<0.01), the membrane potential of hippocampus, the mRNA and protein expression levels of Opa1, Mfn1 and Mfn2 in hippocampus were decreased (<0.01, <0.001) in the model group, while the average fluorescence intensity of ROS was increased, the mRNA and protein expression levels of Drp1, Fis1 and Mff were increased (<0.001). In comparison with the model group, the escape latency was significantly shortened (<0.05), the times of crossing the original platform were increased (<0.05), the membrane potential of the hippocampus, the mRNA and protein expression levels of Opa1, Mfn1 and Mfn2 were increased (<0.05, <0.001) in the moxibustion and medication groups, while the average fluorescence intensity of ROS in the hippocampus, the mRNA and protein expression levels of Drp1, Fis1 and Mff were decreased (<0.001). The average fluorescence intensity of ROS in the moxibustion group was significantly lower than that in the medication group (<0.001). HE staining showed loose arrangement of neurons, disappearance of partial nucleolus, and necrocytosis after modeling, which were relatively milder in both moxibustion and medication groups. CONCLUSIONS: Moxibustion can effectively improve the cognitive function of VD rats, and its mechanism may be related to regulating hippocampal mitochondrial dynamics-related proteins, promoting mitochondrial fusion, inhibiting excessive mitochondrial division, thereby improving the imbalance of mitochondrial dynamics and alleviating mitochondrial dysfunction-related conditions.
目的：探讨艾灸对血管性痴呆（VD）大鼠海马线粒体动力学相关蛋白的影响，以揭示艾灸治疗VD的潜在机制。 方法：将SD大鼠随机分为假手术组、模型组、艾灸组和药物组，每组12只。采用改良双侧颈总动脉结扎法制备VD模型。对“神庭”（GV24）、“百会”（GV20）和“大椎”（GV14）进行温和艾灸20分钟，每周6天，每天1次。药物组大鼠每日口服尼莫地平（12mg/kg）。上述所有干预均进行4周。采用Morris水迷宫试验评估学习和记忆能力。用HE染色观察海马组织病理学变化。通过流式细胞术检测海马组织中线粒体膜电位（Δψm）和活性氧（ROS）水平。分别用PCR或Western blot检测海马中视神经萎缩蛋白1（Opa1）、线粒体融合蛋白1（Mfn1）、线粒体融合蛋白2（Mfn2）、动力相关蛋白1（Drp1）、裂变蛋白1（Fis1）、线粒体裂变因子（Mff）的mRNA和蛋白表达水平。 结果：与假手术组相比，模型组逃避潜伏期延长（<0.01），穿越原平台次数减少（<0.01），海马膜电位、海马中Opa1、Mfn1和Mfn2的mRNA和蛋白表达水平降低（<0.01，<0.001），而ROS平均荧光强度增加，Drp1、Fis1和Mff的mRNA和蛋白表达水平增加（<0.001）。与模型组相比，艾灸组和药物组逃避潜伏期明显缩短（<0.05），穿越原平台次数增加（<0.05），海马膜电位、海马中Opa1、Mfn1和Mfn2的mRNA和蛋白表达水平增加（<0.05，<0.001），而海马中ROS平均荧光强度、Drp1、Fis1和Mff的mRNA和蛋白表达水平降低（<0.001）。艾灸组ROS平均荧光强度明显低于药物组（<0.001）。HE染色显示建模后神经元排列疏松，部分核仁消失，出现坏死细胞，艾灸组和药物组相对较轻。 结论：艾灸可有效改善VD大鼠的认知功能，其机制可能与调节海马线粒体动力学相关蛋白、促进线粒体融合、抑制线粒体过度分裂，从而改善线粒体动力学失衡、减轻线粒体功能障碍相关状况有关。
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