目的 采用慢性不可预知温和应激所致小鼠焦虑模型探究去甲槟榔碱的体内抗焦虑活性及潜在的作用机制。方法 采用旷场、明暗箱和高架十字迷宫等行为学方法评价去甲槟榔碱对焦虑模型小鼠的行为影响;采用酶联免疫法检测小鼠血清皮质酮含量、血清和脑组织中超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT)水平以及脑组织中神经递质和炎症因子水平;Western blotting检测NMDAR、CamkII、Kalirin、Rac的表达。结果 去甲槟榔碱能显著降低模型小鼠在旷场活动中总路程、平均速度,增加小鼠活动次数;显著增加模型小鼠在明暗箱实验中的穿箱次数和明室时间(P<0.05、0.01)。去甲槟榔碱能显著减少模型小鼠在高架十字迷宫实验中进入闭臂次数和闭臂总路程(P<0.01)。去甲槟榔碱20、40 mg/kg组小鼠血清皮质酮(Cort)、MDA显著降低(P<0.01、0.001),去甲槟榔碱各剂量组小鼠SOD、CAT显著增加(P<0.05、0.001)。去甲槟榔碱各剂量组小鼠脑组织MDA含量显著降低(P<0.01、0.001),SOD、CAT含量显著增加(P<0.001)。去甲槟榔碱各剂量组小鼠脑组织5-羟色胺(5-HT)、多巴胺(DA)、去甲肾上腺素(NE)、γ-氨基丁酸(GABA)含量显著增加(P<0.05、0.01、0.001)。去甲槟榔碱40 mg/kg组小鼠肿瘤坏死因子-α(TNF-α)含量显著降低(P<0.05),去甲槟榔碱各剂量组小鼠白细胞介素(IL)-6、IL-1β含量显著降低(P<0.05、0.001)。去甲槟榔碱40 mg/kg组可显著上调NMDARa、p-CamkII/CamkII、Kalirin/β-actin、Rac/β-Actin蛋白表达(P<0.001)。结论 去甲槟榔碱具备显著抗焦虑活性,其作用机制与对抗氧化应激损伤、抑制神经炎性反应、调节神经递质水平以及NMDAR/CamkⅡ/Rac信号通路有关。

Objective To investigate the antianxiety activity of guvacoline in vivo and its potential mechanism induced by chronic and unpredictable mild stress. Methods Behavioral methods such as open field, light and dark box, and elevated cross maze were used to evaluate the effect of norarecoline on the behavior of anxious mice. The content of serum corticosterone, the levels of SOD, MDA, and CAT in serum and brain, and the levels of neurotransmitters and inflammatory factors in brain were detected by enzyme-linked immunoassay. Western blotting detected the expressions of NMDAR, CamkII, Kalirin, and Rac. Resluts Guvacoline can significantly >reduce the total distance and average speed of the model mice in the open field, and increase the number of mouse activities. The penetration times and open chamber time of model mice in light and dark chamber experiment were significantly increased (P < 0.05, 0.01). Guvacoline could significantly reduce the number of times and the total distance of the closed arm in the elevated cross maze experiment (P < 0.01). Serum Cort and MDA in 20 and 40 mg/kg guvacoline groups were significantly decreased (P < 0.01, 0.001), while SOD and CAT were significantly increased in all guvacoline dose groups (P < 0.05, 0.001). The MDA content in the brain tissue of mice in each dose group of guvacoline was significantly reduced (P < 0.01 and 0.001), while the SOD and CAT contents were significantly increased (P < 0.001). The contents of 5-HT, DA, NE, and GABA in brain tissue of mice in guvacoline each dose group were significantly increased (P < 0.05, 0.01, 0.001). The content of TNF-α in guvacoline 40 mg/kg group was significantly decreased (P < 0.05), and the contents of IL-6 and IL-1β in guvacoline dose groups were significantly decreased (P < 0.05, 0.001). The expressions of NMDARa, p-CAMkII/CamkII, Kalirin/β-actin, Rac/β-actin were significantly up-regulated in guvacoline 40 mg/kg group (P < 0.001). Conclusion Guvacoline has significant anti-anxiety activity, and its mechanism is related to anti-oxidative stress injury, inhibition of neuroinflammatory response, regulation of neurotransmitter levels and NMDAR/CamkⅡ/Rac signaling pathway.

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中国农业科学院农产品加工研究所创新工程（CAAS-ASTIP-2023-IFST）；三亚中国农业科学院国家南繁研究院“南繁专项”2022年院院联合攻关项目（YYLH05）；三亚中国农业科学院国家南繁研究院“南繁专项”2023年重点项目（ZDXM2302）