Zhu Sijia, Liu Meijun, Han Shiyu, Zhu Jingyi, Deng Xinmin, Tian Yanyan, Yang Dongdong
Neurology Department, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu 610075, China.
Neurology Department, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu 610075, China.
J Pharm Biomed Anal. 2025 May 15;257:116679. doi: 10.1016/j.jpba.2025.116679. Epub 2025 Jan 19.
The incidence of Parkinson's disease (PD) increases with age. Previous pharmacological studies have shown the potential of Huatan Jieyu Granules (HGs) for the treatment of PD, but the exact mechanisms remain unclear. This study aimed to explore the effects of herbal treatment on PD using mouse models and single-cell sequencing.
In this study, we established in vivo 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD models in mice. Motor function was assessed through behavioral tests. Immunofluorescence was used to examine dopaminergic neuron loss. Single-cell sequencing was performed on mice from the blank, PD model and medication groups. After quality control and dimensionality reduction of the single-cell data, cells were clustered, and different cell types were identified. We then identified the intersection of differentially expressed genes (DEGs1) in the blank and model groups and DEGs2 in the model and medication groups, yielding intersected DEGs. Key drug targets were identified by intersecting these DEGs with the drug targets of active ingredients in TCM. Topological analysis of the PPI network was used to identify key genes. Cell types exhibiting high expression of these genes were designated as key cells. These key cells were subjected to cellular communication analysis and temporal analysis, after which they were classified into subtypes.
HGs significantly improved motor function and prevented dopaminergic neuronal loss in the substantia nigra (SN) of MPTP-treated mice. A total of 34 cell clusters were delineated, with 9 cell types identified, including oligodendrocytes (oligo), neurons, and T cells. We identified 758 intersected DEGs and 13 key drug targets, including Egfr, Ntrk2, Grm5, Htr2c, Bcl2l1. Oligo and neuronal cells were identified as key cells due to higher expression levels of these key genes. In the cellular communication analysis, oligo-neuronal interactions in the blank and model groups, and oligo-OPC and oligo-T cell interactions in the medication group, exhibited the most receptor-ligand interactions. In temporal analysis, both oligo and neuronal cells were differentiated into 9 states, with C1 being the most differentiated.
HGs demonstrate neuroprotective effects in MPTP-treated mice. Using single-cell sequencing, we identified five key genes (Egfr, Ntrk2, Grm5, Htr2c, Bcl2l1) and two key cell types (oligo and neuronal) related to HGs in PD. These findings provided a foundation for understanding the molecular mechanisms by which HGs treat PD.
帕金森病（PD）的发病率随年龄增长而增加。先前的药理学研究表明，化痰解郁颗粒（HGs）具有治疗PD的潜力，但其确切机制尚不清楚。本研究旨在利用小鼠模型和单细胞测序探索中药治疗PD的效果。
在本研究中，我们建立了体内1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）诱导的小鼠PD模型。通过行为测试评估运动功能。采用免疫荧光法检测多巴胺能神经元损失。对空白组、PD模型组和药物治疗组的小鼠进行单细胞测序。对单细胞数据进行质量控制和降维后，对细胞进行聚类，并鉴定不同的细胞类型。然后我们确定了空白组和模型组中差异表达基因（DEGs1）与模型组和药物治疗组中差异表达基因（DEGs2）的交集，得到交集差异表达基因。通过将这些差异表达基因与中药活性成分的药物靶点进行交叉分析，确定关键药物靶点。利用蛋白质-蛋白质相互作用（PPI）网络的拓扑分析确定关键基因。将高表达这些基因的细胞类型指定为关键细胞。对这些关键细胞进行细胞通讯分析和时间分析，然后将它们分类为亚型。
HGs显著改善了MPTP处理小鼠的运动功能，并预防了黑质（SN）中多巴胺能神经元的损失。共划分出34个细胞簇，鉴定出9种细胞类型，包括少突胶质细胞（oligo）、神经元和T细胞。我们确定了758个交集差异表达基因和13个关键药物靶点，包括表皮生长因子受体（Egfr）、神经营养酪氨酸激酶受体2（Ntrk2）、代谢型谷氨酸受体5（Grm5）、5-羟色胺受体2C（Htr2c）、凋亡抑制蛋白（Bcl2l1）。由于这些关键基因的表达水平较高，oligo和神经元细胞被确定为关键细胞。在细胞通讯分析中，空白组和模型组中的oligo-神经元相互作用，以及药物治疗组中的oligo-少突胶质前体细胞（OPC）和oligo-T细胞相互作用，表现出最多的受体-配体相互作用。在时间分析中，oligo和神经元细胞均分化为9种状态，其中C1状态分化程度最高。
HGs在MPTP处理的小鼠中表现出神经保护作用。通过单细胞测序，我们确定了与PD中HGs相关的五个关键基因（Egfr、Ntrk2、Grm5、Htr2c、Bcl2l1）和两种关键细胞类型（oligo和神经元）。这些发现为理解HGs治疗PD的分子机制提供了基础。
