Xu Shihua, Tian Yu, Chen Xiaofei, Zhang Wenjuan, Mei Yingwu, Zhang Qi, Zhang Haifeng
Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, Henan, China.
Technical Research and Development Center, Shanxi Kunming Tobacco Co, Ltd, Taiyuan, Shanxi, China.
Drug Metab Dispos. 2025 Jun;53(6):100084. doi: 10.1016/j.dmd.2025.100084. Epub 2025 Apr 28.
Nicotine withdrawal is often accompanied by anxiety and depression-like behaviors, which are closely linked to neuroinflammation. Theobromine (TB), a bioactive compound found in cocoa, has demonstrated potential anti-inflammatory and neuroprotective properties. This study aimed to quantify the distribution of TB in various tissues-particularly brain regions involved in emotional regulation-and evaluate its efficacy in alleviating nicotine withdrawal-induced emotional disturbances in mice. We first developed a high-efficiency liquid chromatography-tandem mass spectrometry quantitative method for TB and found that its highest concentration was in the blood, followed by the liver, with the lowest levels in the brain. Within the various brain regions, the nucleus accumbens (NAc) exhibited the highest concentration of TB (41.64 ± 17.14 μg/g). Behavioral experiments revealed that TB significantly reduced anxiety and depression-like behaviors induced by nicotine withdrawal. Molecular analyses showed that TB markedly decreased the expression of proinflammatory cytokines (C-C motif chemokine ligand 4, C-C motif chemokine ligand 5, cyclooxygenase-2, toll-like receptor 4, and inducible nitric oxide synthase) in the NAc and lowered levels of interleukin-2, interleukin-6, and tumor necrosis factor-α in both the NAc and serum. In summary, by optimizing liquid chromatography-tandem mass spectrometry technology, we discovered for the first time, to our knowledge, that TB is distributed across nearly all brain regions, with the highest concentration in the NAc. This provides a pharmacological basis for its neuroprotective and mood-regulating effects. TB effectively mitigates anxiety and depression-like behaviors associated with nicotine withdrawal in mice, likely through its anti-inflammatory actions in the NAc and systemic circulation. This study offers compelling evidence for TB as a natural anti-inflammatory and mood-regulating agent, laying the groundwork for developing novel adjunctive treatments for nicotine withdrawal. SIGNIFICANCE STATEMENT: To the authors' knowledge, this study provides the first comprehensive mapping of theobromine (TB) distribution in the brain, specifically focusing on its interactions with common withdrawal and addiction-related pathways, identifying its highest concentration in the nucleus accumbens-a key region for emotional regulation. Demonstrating that TB effectively alleviates nicotine withdrawal-induced anxiety and depression-like behaviors by suppressing neuroinflammation, the findings of this study support the development of TB-based therapies, presenting a novel strategy to aid individuals in overcoming nicotine dependence.
尼古丁戒断通常伴有焦虑和抑郁样行为，这些行为与神经炎症密切相关。可可碱（TB）是可可中发现的一种生物活性化合物，已显示出潜在的抗炎和神经保护特性。本研究旨在量化TB在各种组织中的分布，特别是参与情绪调节的脑区，并评估其减轻小鼠尼古丁戒断诱导的情绪障碍的功效。我们首先开发了一种用于TB的高效液相色谱-串联质谱定量方法，发现其最高浓度存在于血液中，其次是肝脏，脑中浓度最低。在各个脑区中，伏隔核（NAc）的TB浓度最高（41.64±17.14μg/g）。行为实验表明，TB显著减少了尼古丁戒断诱导的焦虑和抑郁样行为。分子分析表明，TB显著降低了NAc中促炎细胞因子（C-C基序趋化因子配体4、C-C基序趋化因子配体5、环氧化酶-2、Toll样受体4和诱导型一氧化氮合酶）的表达，并降低了NAc和血清中白细胞介素-2、白细胞介素-6和肿瘤坏死因子-α的水平。总之，通过优化液相色谱-串联质谱技术，据我们所知，我们首次发现TB分布于几乎所有脑区，NAc中浓度最高。这为其神经保护和情绪调节作用提供了药理学基础。TB有效减轻了小鼠与尼古丁戒断相关的焦虑和抑郁样行为，可能是通过其在NAc和全身循环中的抗炎作用。本研究为TB作为一种天然抗炎和情绪调节剂提供了有力证据，为开发尼古丁戒断的新型辅助治疗奠定了基础。意义声明：据作者所知，本研究首次全面绘制了可可碱（TB）在脑中的分布图，特别关注其与常见戒断和成瘾相关途径的相互作用，确定其在伏隔核（情绪调节的关键区域）中浓度最高。本研究结果表明，TB通过抑制神经炎症有效减轻尼古丁戒断诱导的焦虑和抑郁样行为，支持基于TB的疗法的开发，提出了一种帮助个体克服尼古丁依赖的新策略。
