Zhang Tiange, Gu Jingna, Li Yanlin, Li Huizhen, Yan Can, Wu Lili
Integrative Medicine Research Center, School of Basic Medical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, China.
School of the First Clinical Medical, Guangzhou University of Chinese Medicine, Guangzhou, China.
Eur J Pharmacol. 2025 Jun 26;1003:177860. doi: 10.1016/j.ejphar.2025.177860.
To elucidate the antidepressant effects of Quercetin in a rat model of Chronic Unpredictable Mild Stress (CUMS), explore the underlying mechanisms of Quercetin, and offer novel insights into developing antidepressant agents derived from substances with both medicinal and nutritional applications.
Herein, 75 male rats were sourced and randomly assigned to three groups: Control, CUMS, and Quercetin (50 mg/kg/day). Except for the control group, all rats were exposed to social isolation and chronic stress for eight weeks. The Sucrose Preference Test (SPT), Elevated Plus Maze Test (EPMT), Three-Chamber Social Interaction Test (TCT), and Forced Swimming Test (FST) were used to assess the antidepressant effects of Quercetin on CUMS-exposed rats. Flow cytometry, Immunofluorescence (IF) staining, and Western Blotting (WB) were employed to quantify CD4 T cells and detect their protein expression in the Choroid Plexus (CP), Cerebrospinal Fluid (CSF), plasma, and hippocampus. The IL-1β, IL-6, IL-4, and IL-10 cytokine levels were measured using the Enzyme-Linked Immunosorbent Assay (ELISA). The expressions of ionized calcium-binding adapter molecule 1 (Iba-1), inducible Nitric Oxide Synthase (iNOS), and Arginase-1 in hippocampal tissue were analyzed using IF and WB. Additionally, the CP was examined for ZO-1, CCL5, CCL11, CXCL9, CXCL10, ICAM-1, and VCAM-1 expression through IF. In vitro, HAPI microglial cells were treated with the CSF from the three groups (CON-CSF, CUMS-CSF, and Quercetin-CSF) and cell viability was assessed using the CCK-8 assay. The expressions of Iba-1, iNOS, and Arginase-1 in HAPI microglial cells were also assessed using IF.
As demonstrated by the SPT, FST, EPMT, and TCT behavioral tests, Quercetin significantly improved depressive and anxiety-like behaviors, as well as social deficits in CUMS rats. It also upregulated ZO-1, chemokines (CCL5, CCL11, and CXCL10), and adhesion molecules (ICAM-1) in the CP of CUMS rats. Additionally, Quercetin facilitated moderate recruitment of CD4 T lymphocytes to the CP, suppressed the production of pro-inflammatory cytokines (IL-1β, IL-6), reduced hippocampal microglial cell density, and inhibited M1-type microglial polarization.
Herein, Quercetin moderately promoted the recruitment of CD4 T lymphocytes to the CP, enhanced the Immune Microenvironment (IME) of the CP-CSF-hippocampal axis, and influenced microglial polarization in the hippocampus, highlighting some of the potential mechanisms underlying its antidepressant effects. In light of the current limitations of conventional antidepressants, including suboptimal efficacy and Adverse Reactions (AR), our findings underscore the potential value of Quercetin as an effective antidepressant with both medicinal and nutritional applications.
阐明槲皮素在慢性不可预测轻度应激（CUMS）大鼠模型中的抗抑郁作用，探究槲皮素的潜在作用机制，并为开发兼具药用和营养应用价值的抗抑郁药物提供新见解。
本研究选取75只雄性大鼠，随机分为三组：对照组、CUMS组和槲皮素组（50 mg/kg/天）。除对照组外，所有大鼠均接受8周的社会隔离和慢性应激处理。采用蔗糖偏好试验（SPT）、高架十字迷宫试验（EPMT）、三室社交互动试验（TCT）和强迫游泳试验（FST）评估槲皮素对CUMS大鼠的抗抑郁作用。运用流式细胞术、免疫荧光（IF）染色和蛋白质印迹法（WB）对脉络丛（CP）、脑脊液（CSF）、血浆和海马中的CD4 T细胞进行定量分析，并检测其蛋白表达。采用酶联免疫吸附测定法（ELISA）检测白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、白细胞介素-4（IL-4）和白细胞介素-10（IL-10）细胞因子水平。通过IF和WB分析海马组织中离子钙结合衔接分子1（Iba-1）、诱导型一氧化氮合酶（iNOS）和精氨酸酶-1的表达。此外，通过IF检测CP中紧密连接蛋白1（ZO-1）、趋化因子（CCL5、CCL11、CXCL9、CXCL10）和黏附分子（ICAM-1、VCAM-1）的表达。体外实验中，用三组大鼠的脑脊液（CON-CSF、CUMS-CSF和槲皮素-CSF）处理HAPI小胶质细胞，采用CCK-8法评估细胞活力。同时，通过IF评估HAPI小胶质细胞中Iba-1、iNOS和精氨酸酶-1的表达。
SPT、FST、EPMT和TCT行为学试验表明，槲皮素显著改善了CUMS大鼠的抑郁和焦虑样行为以及社交缺陷。它还上调了CUMS大鼠CP中ZO-1、趋化因子（CCL5、CCL11和CXCL10）和黏附分子（ICAM-1）的表达。此外，槲皮素促进了CD4 T淋巴细胞适度募集至CP，抑制了促炎细胞因子（IL-1β、IL-6）的产生，降低了海马小胶质细胞密度，并抑制了M1型小胶质细胞极化。
本研究中，槲皮素适度促进了CD4 T淋巴细胞向CP的募集，增强了CP-CSF-海马轴的免疫微环境（IME），并影响了海马中的小胶质细胞极化，突出了其抗抑郁作用的一些潜在机制。鉴于传统抗抑郁药目前存在疗效欠佳和不良反应（AR）等局限性，我们的研究结果强调了槲皮素作为一种兼具药用和营养应用价值的有效抗抑郁药的潜在价值。
