南方医科大学学报

南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (7): 1520-1532.doi: 10.12122/j.issn.1673-4254.2026.07.06

• • 上一篇    

化瘀通络灸联合神经干细胞移植对血管性痴呆大鼠Drp1/Mfn蛋白表达及线粒体动力学的影响

尹维帅1(), 李飞2, 曹奕5, 杨琪琪2, 邓则天3, 曹世杰1, 孙永旗1, 毕昕阳1, 梁正鹏1, 伍思嘉1, 王保国4()   

  1. 1.安徽中医药大学第二临床医学院，安徽 合肥 230061
    2.安徽中医药大学第二附属医院 康复二科，安徽 合肥 230061
    4.安徽中医药大学第二附属医院 脑病七科，安徽 合肥 230061
    5.；安徽中医药大学第二附属医院 名医堂，安徽 合肥 230061
    3.安徽农业大学国家重点生物实验室，安徽 合肥 230036
  • 收稿日期:2025-12-08 出版日期:2026-07-20 发布日期:2026-07-20
  • 通讯作者: 王保国 E-mail:1366031873@qq.com;wangbaoguotcm@126.com
  • 作者简介:尹维帅，在读硕士研究生，E-mail: 1366031873@qq.com
  • 基金资助:
    国家青年岐黄学者支持项目(国中医药人教函（2022）256号);第六批全国中医临床优秀人才研修项目(国中医药人教函（2025）256号);安徽省高等学校科学研究重点项目(2023AH050820);安徽中医药大学2024年度临床科研项目（第一批）(2024LC085);安徽省中医药高水平传承人才支持项目(皖中医药发展秘[2024]1号)

Effect of Huayu Tongluo moxibustion combined with neural stem cell transplantation on hippocampal Drp1/Mfn proteins and mitochondrial dynamics in rats with vascular dementia

Weishuai YIN1(), Fei LI2, Yi CAO5, Qiqi YANG2, Zetian DENG3, Shijie CAO1, Yongqi SUN1, Xinyang BI1, Zhengpeng LIANG1, Sijia WU1, Baoguo WANG4()   

  1. 1.Second Clinical Medical College, Anhui University of Chinese Medicine, Hefei 230061, China
    2.Rehabilitation Department II, Second Affiliated Hospital of Anhui University of Chinese Medicine，
    4.Department of Encephalopathy VII, Second Affiliated Hospital of Anhui University of Chinese Medicine，
    5.Famous Doctors Clinic, Second Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230061, China
    3.State Key Laboratory of Biology, Anhui Agricultural University, Hefei 230036, China
  • Received:2025-12-08 Online:2026-07-20 Published:2026-07-20
  • Contact: Baoguo WANG E-mail:1366031873@qq.com;wangbaoguotcm@126.com

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摘要：

目的 探讨化瘀通络灸调控神经干细胞（NSCs）线粒体动力学治疗血管性痴呆（VD）的作用机制。 方法 将72只SPF级SD大鼠随机分为假手术组、VD组、VD+定位注射等量细胞培养基组（VD+CM组）、VD+定位注射NSCs悬液组（VD+NSCs组）、VD+化瘀通络灸组（VD+HYTLM组）、VD+定位注射NSCs悬液+化瘀通络灸组（VD+NSCs+HYTLM组），12只/组。分别于造模后和治疗后，采用Morris水迷宫实验检测大鼠学习、运动能力；TUNEL法、免疫荧光单标法观察海马CA1区脑组织神经元凋亡率及神经干细胞分化情况；苏木精-伊红（HE）染色、免疫荧光共染色检测海马CA1区脑组织神经元存活情况及神经元病理形态；流式细胞仪测定各海马CA1区脑组织线粒体活性氧（ROS）；Western blotting法检测Drp1、Mfn1、Mfn2、Fis1的蛋白表达情况。 结果 与假手术组相比，VD组大鼠逃避潜伏期延长（P<0.01），运动轨迹紊乱；神经元凋亡率上升（P<0.001）；神经干细胞增殖抑制，成熟神经元丢失，新生神经元抑制；ROS平均水平上升（P<0.001）；神经元大面积结构破坏，胞体缩小、胞核固缩，排列疏松，局灶性空洞形成；Drp1、Mfn1、Mfn2 表达下降（P<0.05），Fis1表达上升（P<0.001）。与VD组相比，VD+定位注射等量细胞培养基组无明显变化（P>0.05）。与VD+定位注射等量细胞培养基组比较，VD+NSCs组大鼠逃避潜伏期缩短（P<0.05），运动轨迹趋于集中；神经元凋亡率下降（P<0.05）；神经干细胞增殖增加，成熟神经元数量增多，新生神经元生成增强；ROS平均水平降低（P<0.001）；神经元形态改善，胞体较饱满、胞核固缩减轻，细胞排列稍紧密；Drp1、Mfn1、Mfn2表达上升（P<0.001，P<0.001，P<0.01），Fis1表达下降（P<0.05）。与VD+NSCs组比较，VD+化瘀通络灸组无明显变化（P>0.05）。与VD+化瘀通络灸组比较，VD+NSCs+化瘀通络灸组逃避潜伏期缩短（P<0.001），运动轨迹接近假手术组；神经元凋亡率下降（P<0.01）；神经干细胞增殖、成熟神经元存活及新生神经元生成增强；ROS平均水平下降（P<0.01）；结构完整，胞体饱满，排列紧密；Drp1、Mfn1、Mfn2蛋白表达上升（P<0.001，P<0.01，P<0.01），Fis1表达下降（P<0.05）。 结论 化瘀通络灸法可促进VD大鼠认知功能改善，诱导NSCs分化为功能性神经元，提升神经元存活率并减轻病理损伤。其机制可能通过调节海马区线粒体动力学相关蛋白，促进融合、抑制过度裂变，恢复线粒体动态平衡，从而缓解功能障碍。

关键词: 血管性痴呆, 化瘀通络灸, 神经干细胞, 线粒体动力学, 神经元

Abstract:

Objective To investigate the mechanism by which Huayu Tongluo moxibustion (HYTLM) regulates mitochondrial dynamics of neural stem cells (NSCs) for treatment of vascular dementia (VD). Methods Seventy-two SD rats were randomized equally into 6 groups, including a sham-operated group, a VD model group, and 4 VD groups receiving HYTLM treatment, HYTLM treatment with stereotactic injection of NSC suspension into the hippocampal CA1 region, or injections of NSC suspension or cell culture medium without HYTLM treatment. Morris water maze test was used to assess learning and memory ability of the rats. Hippocampal neuronal apoptosis, NSC differentiation, neuronal survival, histopathological changes, mitochondrial reactive oxygen species (ROS) levels, and hippocampal expressions of Drp1, Mfn1, Mfn2, and Fis1 proteins were evaluated using TUNEL assay, immunofluorescence staining, HE staining, flow cytometry, and Western blotting. Results The rats receiving culture medium injection, similar to the VD rats, exhibited prolonged escape latency and disorganized swimming trajectories in Morris water maze test and had significantly increased neuronal apoptosis, suppressed NSC proliferation, reduced mature neurons, impaired neurogenesis, and increased ROS levels in the hippocampal CA1 region, showing also obvious neuronal injuries, lowered hippocampal Drp1, Mfn1, and Mfn2 expressions and increased Fis1 expression. All these changes were significantly alleviated in VD rats receiving NSC transplantation. HYTLM treatment produced similar effects to NSC transplantation, but their combined treatment further shortened the escape latency of the VD rats, which showed similar swimming patterns to the sham-operated rats, and produced stronger protective effects on the hippocampal neurons. The combined treatment also further reduced ROS levels and improved aberrant expressions of Drp1, Mfn1, Mfn2, and Fis1. Conclusion HYTLM treatment improves cognitive functions of VD rats, promotes NSC differentiation into functional neurons and neuronal survival, and attenuates neuronal injury possibly by regulating hippocampal mitochondrial dynamics, thereby restoring mitochondrial homeostasis and ameliorating functional impairment.

Key words: vascular dementia, Huayu Tongluo moxibustion, neural stem cells, mitochondrial dynamics, neurons