| 摘要: |
| 目的：基于单细胞转录组测序技术探索脑泰方对缺血性卒中大鼠脑组织各细胞亚群的影响。方法：采用改良线栓法构建大鼠脑中动脉阻塞模型，随机分为假手术组、模型组、脑泰方低剂量组、脑泰方中剂量组、脑泰方高剂量组、去铁酮组，每组各10只。药物干预后，采用神经功能评分评估大鼠神经功能恢复情况，采用苏木素-伊红（Hematoxylin-Eosin，HE）染色和普鲁士蓝染色观察大鼠脑组织病理变化及铁沉积情况，利用单细胞转录组测序技术分析模型组和脑泰方高剂量组大鼠缺血侧脑组织细胞亚群的差异，采用Seurat包分析脑泰方干预后各细胞亚群的变化。结果：1）与模型组相比，脑泰方组和去铁酮组大鼠神经功能评分明显改善（P＜0.05），表明其能够改善缺血性卒中后的神经功能障碍。HE染色结果显示，脑泰方组神经元结构破坏较模型组明显减轻，细胞排列整齐，核膜清晰，胞浆染色均匀;普鲁士蓝染色进一步证实，脑泰方可显著减轻缺血侧皮质和海马区的神经元坏死和铁沉积现象，提示其可能通过调控铁代谢缓解铁死亡。2）单细胞转录组测序分析一共鉴定了22个细胞亚群，包括巨噬细胞、Mag+少突胶质细胞、Il1b+小胶质细胞、Spp1+小胶质细胞、中性粒细胞、成纤维细胞、Lgals3+小胶质细胞、Mrc1+巨噬细胞、壁细胞和Nr2f2+星形胶质细胞等。脑泰方干预后，5个小胶质细胞亚群均发生了显著变化。结论：脑泰方可能通过调控C1q high小胶质细胞、Il1b+小胶质细胞和GPR34+小胶质细胞、Lgals3+小胶质细胞与Spp1+小胶质细胞等细胞群的增殖或凋亡发挥治疗缺血性卒中的作用。 |
| 关键词: 缺血性卒中 脑泰方 铁死亡 单细胞转录组学 神经炎症 |
| DOI： |
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| Mechanism of Naotai prescription in treatment of ischemic stroke:A study based on single-cell transcriptomics |
| YANG Kailin,WANG Shanshan,LIU Han |
| (Hunan Provincial Key Laboratory of Disease Model Research for Integrated Traditional Chinese and Western Medicine，Hunan Academy of Chinese Medicine，Changsha 410013，Hunan，China;Key Laboratory of Hunan Province for Prevention and Treatment of Integrated Traditional Chinese and Western Medicine on Cardiocerebral Diseases，School of Integrated Traditional Chinese and Western Medicine，Hunan University of Chinese Medicine，Changsha 410208，Hunan，China) |
| Abstract: |
| Objective：To investigate the effect of Naotai prescription on cell subsets in the brain tissue of rats with ischemic stroke based on single-cell transcriptome sequencing.Methods：The modified suture method was used to establish a rat model of middle cerebral artery occlusion，and then the rats were randomly divided into sham-operation group，model group，deferiprone group，and low-，middle-，and high-dose Naotai prescription groups，with 10 rats in each group.After drug intervention，neurological score was used to assess the recovery of neurological function in rats;HE staining and Prussian blue staining were used to observe pathological changes and iron deposition in brain tissue;single-cell transcriptome sequencing was used to investigate the differences in cell subsets in the ischemic side of brain between the model group and the high-dose Naotai prescription group;Seurat package was used to analyze the changes in cell subsets after intervention with Naotai prescription.Results：Compared with the model group，the Naotai prescription groups and the deferiprone groups had a significant improvement in neurological score (P < 0.05)，suggesting that they could improve neurological dysfunction after ischemic stroke.HE staining showed that compared with the model group，the Naotai prescription groups had significant alleviation of neuronal structural damage，with ordered arrangement of cells，clear nuclear membranes，and uniform staining of cytoplasm，and Prussian blue staining further confirmed that Naotai prescription significantly alleviated neuronal necrosis and iron deposition in the cortex and the hippocampus at the ischemic side of brain，suggesting that Naotai prescription might alleviate ferroptosis by regulating iron metabolism.A total of 22 cell subsets were identified by single-cell transcriptome sequencing，including macrophages，Mag+ oligodendrocytes，Il1b+ microglial cells，Spp1+ microglial cells，neutrophils，fibroblasts，Lgals3+ microglial cells，Mrc1+ macrophages，parietal cells，and Nr2f2+ astrocytes.There were significant changes in 5 microglial cell subsets after intervention with Naotai prescription.Conclusion：Naotai prescription may exert a therapeutic effect on ischemic stroke by regulating the proliferation or apoptosis of the cell subsets such as C1q high microglial cells，Il1b+ microglial cells，GPR34+ microglial cells，Lgals3+ microglial cells，and Spp1+ microglia. |
| Key words: ischemic stroke Naotai prescription ferroptosis single-cell transcriptomics Neuroinflammation |
