Supplementary Material for: Trigonelline through the Attenuation of Oxidative Stress Exerts Antidepressant- and Anxiolytic-Like Effects in a Mouse Model of Maternal Separation Stress

Depression and anxiety are common psychiatric disorders accounting for social and economic burdens. Previous studies have shown that oxidative stress and oxidant/antioxidant imbalance are involved in the pathophysiology of psychiatric disorders. Experiencing early-life adversities (like maternal separation [MS] stress) provoked psychiatric disorders. Trigonelline (TRG) is a pyridine alkaloid that has various pharmacological effects including hypoglycemic, neuroprotective and memory-improving properties. To investigate the antidepressant- and anxiolytic-like effects of TRG focusing oxidative stress, we applied the MS paradigm to male mice at postnatal day (PND) 2–14 (3 h daily, 9–12 a.m.) and investigated the behaviors at PND 45–47. Using valid behavioral tests including a forced swimming test (FST), splash test, open field test (OFT) and elevated plus maze (EPM), we investigated behavioral modifications. Additionally, we examined the effects of MS and TRG treatment on the level of malondialdehyde (MDA), nitric oxide (NO) and also antioxidant capacity in the brain and serum. Our results showed that MS provoked depressive- and anxiety-like behaviors in the FST, OFT, EPM and splash test, which are associated with an increase in MDA and NO levels as well as a decrease in antioxidant capacity in the brain and serum samples. Findings determined that TRG significantly reversed the negative effects of MS on behavior that is accompanied by a decrease in MDA and NO as well as an increase in antioxidant capacity. Findings of the present study showed that beneficial effects of TRG may be, at least partially, mediated via the reduction of oxidative stress and an increase of antioxidant capacity.

抑郁与焦虑是常见的精神障碍，可造成社会与经济负担。既往研究表明，氧化应激及氧化剂/抗氧化剂失衡参与精神障碍的病理生理过程。早期生活逆境（如母婴分离（maternal separation, MS）应激）可诱发精神障碍。葫芦巴碱（Trigonelline, TRG）是一种吡啶类生物碱，具备降糖、神经保护及改善记忆等多种药理学活性。为围绕氧化应激探究TRG的抗抑郁与抗焦虑样作用，我们采用产后第2~14天（postnatal day, PND，每日上午9时至12时，每日3小时）的母婴分离造模范式处理雄性小鼠，并于产后第45~47天开展行为学检测。通过包括强迫游泳实验（forced swimming test, FST）、喷溅实验（splash test）、旷场实验（open field test, OFT）及高架十字迷宫（elevated plus maze, EPM）在内的公认有效行为学测试，我们分析了小鼠的行为学变化。此外，我们还检测了母婴分离与TRG给药对小鼠脑与血清中丙二醛（malondialdehyde, MDA）、一氧化氮（nitric oxide, NO）水平及抗氧化能力的影响。结果显示，母婴分离可诱导小鼠出现抑郁样与焦虑样行为，该现象与脑及血清中MDA、NO水平升高、抗氧化能力下降密切相关。本研究发现，TRG可显著逆转母婴分离所致的行为学异常，同时伴随MDA、NO水平降低及抗氧化能力提升。本研究结果表明，TRG的有益作用至少部分通过减轻氧化应激、提升抗氧化能力实现。