琐琐葡萄黄酮对鱼藤酮致小鼠运动行为障碍的保护作用

摘要/Abstract

摘要：

目的运用鱼藤酮（rotenone，Rot）诱导小鼠运动行为障碍，探究琐琐葡萄黄酮（flavones from vitis vinifera L，VTF）的保护作用及机制。方法将40只C57BL/6雄性小鼠随机分为4组，每组10只，分别为对照（Con）组、Rot组、Rot+VTF组、VTF组。Rot组：连续3周腹腔注射鱼藤酮，剂量为1.5 mg/（kg·d）。Rot+VTF组：先用VTF进行灌胃，剂量为420 mg/（kg·d）；1 h后注射鱼藤酮，剂量为1.5 mg/（kg·d）。VTF组：VTF进行灌胃，剂量为420 mg/（kg.d）；Con组给予等量溶剂。3周后，通过悬挂实验、爬杆实验和步态检测检测小鼠运动行为能力；通过免疫组织化学染色检测小鼠多巴胺能（dopaminergic，DA）神经元损伤情况；通过免疫组织化学染色、Western blot法和q-RTPCR法检测小鼠中脑小胶质细胞活化和M1极化水平。结果与Con相比，Rot组小鼠悬挂实验评分降低、爬杆实验时间延长；与Rot组相比，Rot+VTF组小鼠悬挂实验评分增加、爬杆实验时间缩短，差异均有统计学意义（均P<0.05）。步态检测结果显示，与Con组相比，Rot组小鼠步长缩短，步距变大；与Rot组相比，Rot+VTF组小鼠步长增加，步距缩短，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Con组相比，Rot组小鼠中脑酪氨酸羟化酶（tyrosine hydroxylase-positive，TH）免疫反应阳性神经元明显减少、离子化钙结合适配分子1（ionized calcium-binding adapter molecule1，IBA1）光密度值升高；与Rot组相比，Rot+VTF组TH免疫反应阳性神经元增加、IBA1光密度值降低，差异均有统计学意义（均P<0.05）。Western blot法和q-RTPCR法检测结果显示，与Con组相比，Rot组小鼠中脑IBA1蛋白表达水平和诱导型一氧化氮合成酶（inducible nitric oxide synthase，iNOS）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素（interleukin，IL）-1β和IL-6基因表达水平升高；与Rot组相比，Rot+VTF组中脑IBA1蛋白表达水平和iNOS、TNF-α、IL-1β和IL-6基因表达水平降低，差异均有统计学意义（均P<0.05）。结论 VTF可能通过抑制小鼠中脑小胶质细胞活化和M1极化介导的炎性反应，进而减轻鱼藤酮引发的DA神经元损伤，最终改善运动行为障碍。

关键词: 琐琐葡萄, 黄酮, 鱼藤酮, 多巴胺能神经元, 运动行为障碍, 小胶质细胞

Abstract:

Objective To explore the protective effect and mechanism of flavones from vitis vinifera L（VTF） on motor behavior disorders in mice induced by rotenone. Method Totally 40 C57BL/6 male mice were randomly divided into four groups，with 10 mice in each group：control（Con） group，Rot group，Rot+VTF group，and VTF group. Rot group：rotenone was injected intraperitoneally for 3 consecutive weeks at a dose of 1.5 mg/（kg·d）. Rot+VTF group：mice were gavaged with VTF at a dose of 420 mg/（kg·d） first，and then injected with rotenone at a dose of 1.5 mg/（kg·d） after 1 hour. VTF group：VTF was administered by gavage at 420 mg/（kg·d）. The Con group：mice were injected with equal amounts of solvent. After 3 weeks，the motor behavior ability of mice was detected through suspension experiments，pole climbing experiments，and gait detection. The damage of dopaminergic neurons in mice were detected through immunohistochemical staining. Western blot and q-RTPCR were used to detect the microglia activation and M1 polarization levels in the midbrain. Results Compared with the Con group，the suspension test scores of mice in the Rot group decreased and the pole climbing test time was prolonged. Compared with the Rot group，the Rot+VTF group showed an increase in suspension test scores and a decrease in pole climbing test time，with statistically significant differences（all P<0.05）. The gait detection results showed that compared with the Con group，the stride length of the Rot group mice was shortened and the stride distance was increased. Compared with the Rot group，the mice in the Rot+VTF group showed an increase in stride length and a decrease in stride distance，with statistically significant differences（all P<0.05）. The immunohistochemical staining results showed that compared with the Con group，the number of tyrosine hydroxylase-positive dopaminergic neurons（TH+ cells） in the Rot group mice was significantly reduced，and the ionized calcium-binding adapter molecule1（IBA1） optical density value was increased in the midbrain. Compared with the Rot group，the number of TH+ cells increased and the IBA1 optical density value decreased in the Rot+VTF group，with statistically significant differences（all P<0.05）. The results of Western blot and q-RTPCR detection showed that compared with the Con group，the expression levels of IBA1，inducible nitric oxide synthase（iNOS），tumor necrosis factor-α（TNF-α），interleukin（IL）-1β and IL-6 in the midbrain of mice in the Rot group were increased. Compared with the Rot group，the expression levels of IBA1，iNOS，TNF-α，IL-1β and IL-6 in Rot+VTF group were decreased，with statistically significant differences（all P<0.05）. Conclusion VTF may alleviate the damage to dopaminergic neurons in the midbrain of mice induced by rotenone by inhibiting the microglia activation and M1 polarization-mediated inflammatory response，thereby improving dyskinesia in mice.

Key words: Vitis vinifera L, Flavone, Rotenone, Dopaminergic neurons, Dyskinesia, Microglia

中图分类号:

R114

孙威, 江阿沛. 琐琐葡萄黄酮对鱼藤酮致小鼠运动行为障碍的保护作用. [J]职业与健康, 2026, 42(14): 1924-1929.

SUN Wei, JIANG Apei. Protective effect of flavones from vitis vinifera L on motor behavior disorders induced by rotenone in mice. [J]OCCUPATION AND HEALTH, 2026, 42(14): 1924-1929.

图/表 6

图1 悬挂实验结果注：Con—对照组；Rot—鱼藤酮组；Rot+VTF—鱼藤酮+琐琐葡萄黄酮组；VTF—琐琐葡萄黄酮组；aP<0.05。

图2 爬杆实验结果注：Con—对照组；Rot—鱼藤酮组；Rot+VTF—鱼藤酮+琐琐葡萄黄酮组；VTF—琐琐葡萄黄酮组；aP<0.05。

图3 VTF对鱼藤酮致步态异常的调控作用注：A—左前肢步长；B—右前肢步长；C—左后肢步长；D—右后肢步长；E—前肢步距；F—后肢步距；Con—对照组；Rot—鱼藤酮组；Rot+VTF—鱼藤酮+琐琐葡萄黄酮组；VTF—琐琐葡萄黄酮组；aP<0.05。

图4 VTF对鱼藤酮致DA神经元损伤的调控作用注：A—运用TH抗体对中脑进行免疫组织化学染色；B—统计分析结果；Con—对照组；Rot—鱼藤酮组；Rot+VTF—鱼藤酮+琐琐葡萄黄酮组；VTF—琐琐葡萄黄酮组；aP<0.05。

图5 VTF对鱼藤酮致小胶质细胞活化的调控作用注：A—IBA1免疫组织化学染色结果；B—IBA1光密度值统计分析结果；C—IBA1蛋白表达；D—IBA1蛋白表达统计分析结果；Con—对照组；Rot—鱼藤酮组；Rot+VTF—鱼藤酮+琐琐葡萄黄酮组；VTF—琐琐葡萄黄酮组；aP<0.05。

图6 VTF对鱼藤酮致M1极化的调控作用注：A—iNOS基因表达；B—TNF-α基因表达；C—IL-6基因表达；D—IL-1β基因表达；Con—对照组；Rot—鱼藤酮组；Rot+VTF—鱼藤酮+琐琐葡萄黄酮组；VTF—琐琐葡萄黄酮组；aP<0.05。

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