二氢杨梅素通过抑制LCN2/NLRP3轴促进星形胶质细胞A2型极化抑制神经元铁死亡改善帕金森病小鼠抑郁样行为OA
Dihydromyricetin ameliorates depressive-like behaviors in Parkinson's disease mice by promoting A2 astrocyte polarization and inhibiting neuronal ferroptosis via suppression of the LCN2/NLRP3 axis
目的 抑郁样行为是帕金森病(Parkinson's disease,PD)中最常见的非运动症状之一,严重影响患者的生活质量.二氢杨梅素(dihydromyricetin,DHM)是一种天然黄酮类化合物,具有一定神经保护作用,但其是否能够改善PD抑郁样行为及其分子机制尚不明确.本研究拟探究DHM改善PD抑郁样行为的潜在作用机制,并筛选和验证其可能调控的关键分子靶点.方法 将32只7周龄雄性C57BL/6J小鼠[体质量(24.5±1.5)g]按随机数字表法分为4组(n=8):对照(Control)组、模型(PD)组、阳性对照美多芭干预(PD+Madopar)组和DHM干预(PD+DHM)组.除Control组外,其余各组采用腹腔注射 1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride,MPTP)建立亚急性PD小鼠模型.采用转棒、爬杆、悬挂实验评估小鼠运动能力,采用开放旷场、强迫游泳、悬尾实验检测小鼠抑郁样行为.基于网络药理学、数据集挖掘和分子对接技术预测DHM治疗PD的关键靶点.利用Western blotting和免疫荧光等技术检测DHM对突触可塑性、星形胶质细胞极化和铁死亡的影响.进一步通过细胞条件培养基实验以验证DHM改善PD抑郁样行为的预测机制.结果 与PD组相比,PD+DHM 组小鼠在转棒实验中的停留时间以及悬挂实验中的支撑时间显著延长(P<0.001),爬杆实验中的下杆时间显著缩短(P=0.005 5),旷场实验中的总路程明显增多(P<0.001),平均速度加快(P<0.001),中心活动距离和活动时间显著增加(P<0.01),强迫游泳和悬尾实验中,小鼠的平均不动时间明显缩短(P<0.01).网络药理学和分子对接确定脂质运载蛋白2(lipocalin 2,LCN2)/NOD 样受体热蛋白结构域相关蛋白 3(NOD-like receptor thermal protein domain associcated protein 3,NLRP3)轴为DHM的潜在作用靶点.与PD组相比,DHM干预后逆转了神经元的丢失和尼氏小体的减少,并上调了脑源性神经营养因子(brain derived neurotrophic factor,BDNF)、突触蛋白I(synapsin I,SYN1)、突触后密度蛋白95(postsynaptic density 95,PSD95)等突触可塑性指标的表达(P<0.01).DHM干预后LCN2、NLRP3和星形胶质细胞A1型标志物补体C3(complement component 3,C3)的表达与 PD 组 相 比均显著下调(P<0.001),A2型标志物S100钙结合蛋白A10(S100 calcium binding protein A10,S100A10)的表达显著增加(P<0.01).免疫荧光显示,与PD组相比,PD+DHM组小鼠额叶皮层GFAP/C3阳性细胞数目显著减少而GFAP/S100A10细胞数目显著增加(P<0.05).此外,与PD组相 比,PD+DHM组小鼠额叶皮层组织中酰基辅酶A合成酶长链家族成员4(acyl-CoA synthetase long chain family member 4,ACSL4)、转铁蛋白受体(transferrin receptor,TFRC)表达显著降低(P<0.01),GPX4、SLC7A11蛋白的表达显著升高(P<0.001).细胞实验进一步证实DHM及NLRP3抑制剂MCC950预处理的条件培养基逆转了多巴胺能神经元铁死亡相关指标的异常表达(P<0.01).结论 DHM可能通过抑制LCN2/NLRP3轴促进星形胶质细胞向A2型极化,缓解星形胶质细胞极化失衡与神经元铁死亡,进而改善PD小鼠抑郁样行为.
Objective Depressive-like behaviors is one of the most prevalent non-motor symptoms in Parkinson's disease(PD),severely compromising patients'quality of life.Dihydromyricetin(DHM),a natural flavonoid,exhibits neuroprotective effects,but its ability to ameliorate PD-related depressive-like behaviors and the underlying mechanisms remain unclear.This study aimed to investigate DHM's mechanism for improving depressive-like behaviors in PD and identify its key molecular targets.Methods Thirty-two 7 weeks old male C57BL/6J mice(weighting 24.5±1.5 g)were randomly divided into 4 groups(n=8):Control,PD model,PD+Madopar(positive control),and PD+DHM.Except the control group,all groups received intraperitoneal 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)to establish subacute PD models.Motor function was assessed via rotarod,pole and wire hang tests;depressive-like behaviors were evaluated using the open field,forced swimming,and tail suspension tests.Network pharmacology,dataset mining,and molecular docking predicted DHM's key targets for PD treatment.Western blotting and immunofluorescence detected DHM's effects on synaptic plasticity,astrocyte polarization,and ferroptosis.Conditioned medium experiments further validated the predicted mechanisms.Results Compared with the PD group,PD+DHM mice showed significantly prolonged rotarod duration and suspension time(P<0.001),reduced pole descent time(P=0.005 5),increased total distance(P<0.001)and velocity(P<0.001)in open field test,with higher central activity distance/time ratios(P<0.01),and shorter immobility times in forced swimming and suspension tests(P<0.01).Network pharmacology and molecular docking identified the Lipocalin-2(LCN2)/NOD-like receptor thermal protein domain associcated protein 3(NLRP3)axis as DHM's potential target.DHM intervention reversed neuronal loss and Nissl body reduction while upregulating synaptic plasticity markers BDNF,SYN1 and PSD95,compared with the PD group(P<0.01).DHM significantly downregulated LCN2,NLRP3,and A1-astrocyte marker complement component 3(C3;P<0.001),but upregulated A2-marker S100 calcium binding protein A10(S100A10;P<0.01).Compared with the PD gruop,immunofluorescence revealed reduced GFAP/C3-positive cells and increased GFAP/S100A10 cells in the prefrontal cortex of PD+DHM group(P<0.05).Additionally,DHM decreased acyl-CoA synthetase long chain family member 4(ACSL4)and Transferrin receptor(TFRC)expression(P<0.001),while increasing GPX4 and SLC7A11 levels(P<0.001)in the prefrontal cortex.Conditioned medium experiments confirmed that DHM and NLRP3 inhibitor MCC950 reversed abnormal ferroptosis-related changes in dopaminergic neurons(P<0.01).Conclusion DHM may ameliorate depressive-like behaviors in PD mice by inhibiting the LCN2/NLRP3 axis which promotes A2-astrocyte polarization,mitigates imbalance in astrocyte polarization and alleviates neuronal ferroptosis.
吕梦林;刘小倩;张宝文;寇现娟
武汉体育学院运动医学院,湖北武汉武汉体育学院运动医学院,湖北武汉武汉体育学院运动医学院,湖北武汉武汉体育学院运动医学院,湖北武汉
医药卫生
二氢杨梅素帕金森病铁死亡星形胶质细胞
dihydromyricetinParkinson's diseaseferroptosisastrocytes
《陆军军医大学学报》 2026 (10)
1339-1352,14
湖北省自然科学基金中医药创新发展联合基金重点项目(2024AFD242)湖北省高校优秀中青年科技创新团队项目(T2024019) Supported by the Key Project of Traditional Chinese Medicine Joint Fund for Innovation and Development of Natural Science Foundation of Hubei Province(2024AFD242)and the Program for Excellent Young and Middle-Aged Science and Technology Innovation Team Program for Higher Education Institutions of Hubei Province(T2024019).
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