目的 采用慢性不可預(yù)知溫和應(yīng)激所致小鼠焦慮模型探究去甲檳榔堿的體內(nèi)抗焦慮活性及潛在的作用機(jī)制。方法 采用曠場(chǎng)、明暗箱和高架十字迷宮等行為學(xué)方法評(píng)價(jià)去甲檳榔堿對(duì)焦慮模型小鼠的行為影響;采用酶聯(lián)免疫法檢測(cè)小鼠血清皮質(zhì)酮含量、血清和腦組織中超氧化物歧化酶(SOD)、丙二醛(MDA)、過氧化氫酶(CAT)水平以及腦組織中神經(jīng)遞質(zhì)和炎癥因子水平;Western blotting檢測(cè)NMDAR、CamkII、Kalirin、Rac的表達(dá)。結(jié)果 去甲檳榔堿能顯著降低模型小鼠在曠場(chǎng)活動(dòng)中總路程、平均速度,增加小鼠活動(dòng)次數(shù);顯著增加模型小鼠在明暗箱實(shí)驗(yàn)中的穿箱次數(shù)和明室時(shí)間(P<0.05、0.01)。去甲檳榔堿能顯著減少模型小鼠在高架十字迷宮實(shí)驗(yàn)中進(jìn)入閉臂次數(shù)和閉臂總路程(P<0.01)。去甲檳榔堿20、40 mg/kg組小鼠血清皮質(zhì)酮(Cort)、MDA顯著降低(P<0.01、0.001),去甲檳榔堿各劑量組小鼠SOD、CAT顯著增加(P<0.05、0.001)。去甲檳榔堿各劑量組小鼠腦組織MDA含量顯著降低(P<0.01、0.001),SOD、CAT含量顯著增加(P<0.001)。去甲檳榔堿各劑量組小鼠腦組織5-羥色胺(5-HT)、多巴胺(DA)、去甲腎上腺素(NE)、γ-氨基丁酸(GABA)含量顯著增加(P<0.05、0.01、0.001)。去甲檳榔堿40 mg/kg組小鼠腫瘤壞死因子-α(TNF-α)含量顯著降低(P<0.05),去甲檳榔堿各劑量組小鼠白細(xì)胞介素(IL)-6、IL-1β含量顯著降低(P<0.05、0.001)。去甲檳榔堿40 mg/kg組可顯著上調(diào)NMDARa、p-CamkII/CamkII、Kalirin/β-actin、Rac/β-Actin蛋白表達(dá)(P<0.001)。結(jié)論 去甲檳榔堿具備顯著抗焦慮活性,其作用機(jī)制與對(duì)抗氧化應(yīng)激損傷、抑制神經(jīng)炎性反應(yīng)、調(diào)節(jié)神經(jīng)遞質(zhì)水平以及NMDAR/CamkⅡ/Rac信號(hào)通路有關(guān)。

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.

R285

中國農(nóng)業(yè)科學(xué)院農(nóng)產(chǎn)品加工研究所創(chuàng)新工程（CAAS-ASTIP-2023-IFST）；三亞中國農(nóng)業(yè)科學(xué)院國家南繁研究院“南繁專項(xiàng)”2022年院院聯(lián)合攻關(guān)項(xiàng)目（YYLH05）；三亞中國農(nóng)業(yè)科學(xué)院國家南繁研究院“南繁專項(xiàng)”2023年重點(diǎn)項(xiàng)目（ZDXM2302）