目的 研究杨梅酮对慢性不可预测性温和应激（CUMS）模型小鼠抑郁样行为的影响及潜在作用机制。方法 56只C57BL/6J雄性小鼠随机分为4组：对照组（Con）、CUMS模型组（CUMS）、杨梅酮干预组[Myr,100 mg/（kg·bw）]、氟西汀阳性对照组（FLX）,后两组小鼠每日接受刺激前分别以Myr和FLX灌胃干预;8 w后,采用旷场、悬尾、高架十字迷宫、糖水偏好试验评价小鼠的抑郁样行为,观察海马组织病理学形态,对海马进行microRNA（miRNA）高通量测序,并测定海马神经递质和炎症因子水平,筛选海马靶向Nlrp3的相关miRNA、NLRP3炎症信号通路相关基因和蛋白的表达水平。结果 Myr显著改善CUMS小鼠的抑郁样行为,表现为小鼠在旷场试验中心区域的停留时间和移动距离显著增加,静止不动时间降低,开放臂停留时间和进入次数显著增加,糖水偏好指数显著增加（P<0.05）;同时,Myr干预组海马CA1、CA3和DG区神经元排列较为整齐,5-羟色胺和多巴胺水平显著增加（P<0.05）;与Con组相比,CUMS导致17个miRNA显著变化,Myr显著调节15个miRNA的表达,可显著上调CUMS诱导的miR-223和miR-30a表达下降,抑制靶基因NLRP3和下游Cleaved caspase-1（p20）的基因及蛋白表达,并显著降低白细胞介素-1β（interleukin-1β,IL-1β）、白细胞介素-18（interleukin-18,IL-18）和肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）的水平（P<0.05）。结论 Myr通过调节靶向作用于NLRP3的miR-223和miR-30a的表达,抑制海马NLRP3/Caspase-1信号通路的激活,抑制炎症反应,改善小鼠抑郁样行为。
Abstract
Objective To study the effect of myricetin and its underlying mechanism on depressive-like behaviors in mice with chronic unpredictable mild stress. Methods Fifty-six male C57BL/6J mice were randomly divided into four groups: control group (Con), CUMS model group (CUMS), myricetin intervention group (Myr, 100 mg/(kg∙bw) myricetin), and fluoxetine intervention group (FLX). The mice received oral gavage of myricetin and fluoxetine prior to daily stress. Eight weeks later, open-field, tail suspension, elevated plus maze, and sucrose preference tests were used to assess the depressive-like behaviors. The hippocampal histopathological morphology, microRNA (miRNA) expressions, and the neurotransmitters and inflammatory cytokines were measured. Additionally, the relative expression of miRNA targeting Nlrp3, as well as genes and proteins associated with the NLRP3 inflammatory signaling pathway were determined. Results Myricetin significantly improved depressive-like behaviors in CUMS mice, manifested as significantly increased time spent and distance traveled in the central area of the open-field test, reduced immobility time, increased time spent and entries into the open arms, and elevated sucrose preference index (P<0.05). Concurrently, the Myr group exhibited more orderly neuronal arrangement in hippocampal CA1, CA3, and DG regions, along with significantly elevated 5-hydroxytryptamine and dopamine levels. Compared with the Con group, CUMS induced significant alterations in 17 miRNAs expression, while myricetin significantly regulated the expression of 15 miRNAs compared to the CUMS group. Myricetin significantly upregulated the down-regulated expression of miR-223 and miR-30a induced by CUMS to inhibit the expression of NLRP3 and its downstream Cleaved caspase-1 (p20), and reduced interleukin-1β (IL-1β), interleukin-18 (IL-18), and tumor necrosis factor-α (TNF-α) levels (P<0.05). Conclusion Myricetin inhibit the activation of the hippocampal NLRP3/Caspase-1 signaling pathway and suppress inflammation, by regulating the expression of NLRP3-targeted miR-223 and miR-30a and thereby improves depressive-like behaviors in mice.
关键词
抑郁 /
杨梅酮 /
miRNA /
NLRP3 /
小鼠
Key words
depression /
myricetin /
microRNA /
NLRP3 /
mouse
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基金
中国博士后基金（No.2017M620191）; 江苏省中医药学会科研项目（NO.ZXFZ2024094）
