湖南中医药大学学报2025，Vol.45Issue(12)：2271-2279,9.DOI:10.3969/j.issn.1674-070X.2025.12.005
百合知母汤通过改善线粒体功能与自噬保护秀丽隐杆线虫多巴胺能神经元的机制研究
Mechanism of Baihe Zhimu Decoction in protecting dopaminergic neurons in Caenorhabditis elegans via enhancement of mitochondrial function and autophagy
摘要
Abstract
Objective To investigate the protective effects of Baihe Zhimu Decoction(BHZMD)on dopaminergic neurons(DNs)in Caenorhabditis elegans(C.elegans).Methods Synchronized C.elegans worms were randomly divided into blank control group,model group[exposed to 400 μmol·L-1 rotenone(ROT)],BHZMD group(treated with 6.7 mg·mL-1 BHZMD solution+400 μmol·L-1 ROT),and fluoxetine group(treated with 100 μmol·L-1 fluoxetine solution+400 μmol·L-1 ROT).The optimal concentration of ROT was determined based on the fluorescence intensity of green fluorescent protein(GFP)and the degree of DN deformity of the worms.The optimal concentration of BHZMD was selected by assessing worm head thrash frequency.Dopaminergic neuronal damage was evaluated in the worms with GFP-labeled DNs(BZ555).Mitochondrial function was analyzed using worms expressing fluorescent markers for electron transport chain(ETC)complexes(CB7272)to monitor ETC recovery.Mitochondrial membrane potential was measured using a fluorescence probe.Survival analysis was performed in a juglone-induced oxidative stress model.Intracellular reactive oxygen species(ROS)levels were measured using the DCFH-DA probe.The expression of pink-1 was examined in autophagy-related gene-labeled worms(RB2547)via immunofluorescence.Results The optimal modeling concentrations of ROT and BHZMD were determined to be 400 μmol·L-1 and 6.7 mg·mL-1,respectively.Compared with the blank control group,the model group exhibited significantly decreased GFP fluorescence signals of DNs(P<0.01)and significantly increased degree of DN deformity(P<0.01).Furthermore,the GFP intensity of the mitochondrial ETC complexes was significantly reduced(P<0.01),along with decreased ATP levels(P<0.01),declined mitochondrial membrane potential(P<0.01),lower worm survival rate(P<0.01),elevated ROS levels(P<0.01),and weakened pink-1 fluorescence signals(P<0.05).Compared with the model group,the BHZMD group exhibited higher DN GFP fluorescence signals(P<0.05);both the BHZMD and fluoxetine groups demonstrated reduced degree of DN deformity(P<0.01),enhanced ETC complex GFP intensity(P<0.01),elevated ATP levels(P<0.01),improved mitochondrial membrane potential(P<0.01),higher survival rates(P<0.01),decreased ROS levels(P<0.01),and intensified pink-1 fluorescence signals(P<0.01).Conclusion BHZMD may protect DNs in C.elegans by ameliorating mitochondrial dysfunction,enhancing antioxidant capacity,reducing ROS levels,and upregulating pink-1 expression.关键词
百合知母汤/秀丽隐杆线虫/氧化应激/线粒体功能/多巴胺能神经元Key words
Baihe Zhimu Decoction/Caenorhabditis elegans/oxidative stress/mitochondrial function/dopaminergic neuron分类
医药卫生引用本文复制引用
陈桑妮,吴梅,何薇,孟盼,李丹丹..百合知母汤通过改善线粒体功能与自噬保护秀丽隐杆线虫多巴胺能神经元的机制研究[J].湖南中医药大学学报,2025,45(12):2271-2279,9.基金项目
国家自然科学基金项目(82474468) （82474468）
湖湘青年英才项目(2024RC3200) （2024RC3200）
湖南省卫生健康委员会高层次人才项目(青年骨干)(20240304118) （青年骨干）
长沙市杰出创新青年培养计划(kq2406017). （kq2406017）
