Raw data of NTS-BLA of heroin withdrawal mice.

Anxiety and depression significantly contribute to heroin relapse, and addressing these issues could lower relapse rates. The basolateral amygdala (BLA) and nucleus tractus solitarius (NTS) are involved in regulating these emotions, but the molecular mechanisms during heroin withdrawal are not yet understood. Subcutaneous injection of heroin into C57BL/6J mice to simulate chronic dependence, withdrawal, and Exendin-4 treatment. Assess anxiety and depression-like behaviors using open field test (OFT), elevated plus maze (EPM), forced swimming test (FST), and tail suspension test (TST). Analyze neuronal and protein expression changes in the BLA brain area with Western blotting (WB) and immunofluorescence staining. Heroin dependence reduces glutamatergic neurons in BLA without affecting anxiety and depression-like behaviors, due to the inhibitory effect of heroin reward. During withdrawal, GLP-1 secretion by the NTS rises, increasing c-Fos and GLP-1 receptor expression in glutamatergic neurons of BLA, linked to heightened anxiety but not depression. A 7-day treatment with Exendin-4 (2 µg/kg) alleviates anxiety in withdrawal mice by downregulating GLP-1 signaling in the NTS-BLA circuit, indicating GLP-1’s role in regulating anxiety during heroin withdrawal. GLP-1 receptors within BLA may serve as molecular targets for modulating emotional states, thereby offering empirical support for strategies aimed at preventing heroin relapse.焦虑与抑郁显著促进海洛因复吸，针对此类情绪问题的干预或可降低复吸率。基底外侧杏仁核（basolateral amygdala, BLA）与孤束核（nucleus tractus solitarius, NTS）参与情绪调控，但海洛因戒断过程中的分子机制尚未阐明。本研究通过向C57BL/6J小鼠皮下注射海洛因以模拟慢性海洛因依赖、戒断状态，并给予艾塞那肽（Exendin-4）干预。采用旷场实验（open field test, OFT）、高架十字迷宫（elevated plus maze, EPM）、强迫游泳实验（forced swimming test, FST）及悬尾实验（tail suspension test, TST）评估小鼠的焦虑及抑郁样行为；通过蛋白质免疫印迹（Western blotting, WB）与免疫荧光染色，分析BLA脑区的神经元及蛋白表达变化。实验结果显示：海洛因依赖可降低BLA内的谷氨酸能神经元数量，但未显著影响小鼠的焦虑及抑郁样行为，这或与海洛因奖赏的抑制效应有关。戒断阶段，NTS分泌的胰高血糖素样肽-1（GLP-1）水平升高，可增加BLA内谷氨酸能神经元的c-Fos蛋白与GLP-1受体表达，该变化与焦虑水平升高相关，但与抑郁样行为无关。每日给予2 μg/kg的Exendin-4，连续干预7天，可通过下调NTS-BLA环路中的GLP-1信号通路，缓解戒断小鼠的焦虑症状，表明GLP-1在海洛因戒断期的情绪调控中发挥关键作用。BLA内的GLP-1受体或可作为调控情绪状态的分子靶点，为预防海洛因复吸的干预策略提供实验依据。