中国实验方剂学杂志2026，Vol.32Issue(11)：56-67,12.DOI:10.13422/j.cnki.syfjx.20260124
清热解毒法通过保护内皮-周细胞并抑制巨噬细胞迁移协同减轻缺血性中风损伤
Heat-clearing and Toxin-removing Method Reduces Ischemic Stroke Injury by Protecting Endothelial-pericyte and Inhibiting Macrophage Migration
摘要
Abstract
Objective:To investigate the regulatory effects of Huanglian Jiedutang(HLJDT)on immune cell migration,blood-brain barrier protection,and cellular functional recovery in a model of ischemic stroke.Methods:A transient middle cerebral artery occlusion(tMCAO)model was established in mice to induce ischemic stroke.Cerebral blood flow and neurological function were evaluated using laser speckle imaging and neurological deficit scoring.Histopathological damage in brain tissues was assessed by hematoxylin-eosin(HE)and Nissl staining.Mice were divided into a sham group,a model group,an HLJDT group,and a Ginkgo biloba extract(GBE)group.After one week of acclimatization,intragastric administration was initiated.The sham and model groups received normal saline,the HLJDT group received HLJDT at 1.82 g·kg-1,and the GBE group received GBE at 0.432 g·kg-1.Administration was continued for 5 consecutive days,and the tMCAO model was established after the final dose on day 6.Single-cell RNA sequencing was performed on brain tissues and peripheral immune cells.UMAP and odds ratio(OR)indices were used to analyze cell distribution.Differential expression analysis was conducted to evaluate the effects of HLJDT on endothelial cells,pericytes,and macrophages,combined with CellChat and decoupler to analyze cell-cell communication and transcription factor regulation.Finally,PCR and ELISA were used to validate the mRNA and protein expression of relevant genes.Results:Compared with the sham group,the model group showed significantly increased neurological deficit scores(P＜0.01)and significantly decreased cerebral blood flow(P＜0.01),accompanied by cortical structural disorder,aggravated cytoplasmic vacuolization,and increased numbers of Nissl bodies.Compared with the model group,both the HLJDT and GBE groups exhibited significantly reduced neurological deficit scores(P＜0.01)and markedly improved cerebral blood flow(P＜0.01),along with amelioration of cortical structural disorder,alleviated cytoplasmic vacuolization,and reduced numbers of Nissl bodies.Single-cell analysis showed that HLJDT protected endothelial cells and pericytes by preventing their reduction,restored the expression of functional genes in these cells(e.g.,PECAM1 and NOS3),and downregulated the expression of chemokines and adhesion-related factors(e.g.,CCL2 and CXCL2).In macrophages,HLJDT reduced their recruitment to the central nervous system and downregulated the expression of chemokine receptors and inflammatory factors(e.g.,IL-6,CCR2,and CXCR2).Cell-cell communication analysis further indicated that HLJDT,through the above mechanisms,alleviated damage to pericytes and endothelial cells,reduced their recruitment of macrophages,and decreased ligand-receptor interactions in chemokine signaling pathways(including CCL,CXCL,and CSF3)between pericytes/endothelial cells and macrophages,thereby preventing secondary injury.Compared with the sham group,the model group showed significantly upregulated mRNA expression levels of IL-1β,IL-6,TNF-α,CCL2,CXCL2,and CSF3(P＜0.01),while mRNA expression levels of endothelial-and pericyte function-related genes(RGS5,PECAM1,VEGFB,and NOS3)were significantly downregulated(P＜0.01).In contrast,compared with the model group,the HLJDT and GBE groups exhibited significantly decreased mRNA expression levels of IL-1β,IL-6,TNF-α,CCL2,CXCL2,and CSF3(P＜0.01),and significantly increased expression of RGS5,PECAM1,VEGFB,and NOS3(P＜0.01).At the protein level,compared with the sham group,the model group showed significantly increased expression of IL-1β,IL-6,and TNF-α(P＜0.01),whereas these protein levels were significantly reduced in the HLJDT and GBE groups compared with the model group(P＜0.01).Conclusion:HLJDT reduces neuronal damage in ischemic stroke by protecting endothelial cells and pericytes,while inhibiting their interaction with macrophages,thereby mitigating secondary injury in the central nervous system.关键词
黄连解毒汤/缺血性脑卒中/单细胞转录组/中枢-外周互作Key words
Huanglian Jiedutang/ischemic stroke/single-cell transcriptomics/central-peripheral interaction分类
医药卫生引用本文复制引用
孙资金,王雪茜,程发峰,张浩嘉,王凯,王昭懿,宋粼静,徐文秀,吉静,李长香,王庆国..清热解毒法通过保护内皮-周细胞并抑制巨噬细胞迁移协同减轻缺血性中风损伤[J].中国实验方剂学杂志,2026,32(11):56-67,12.基金项目
国家自然科学基金项目(U21A20400) （U21A20400）
