Chaihu-Shugan-San Alleviates Post-Stroke Depression in Mice: Mechanistic Insights into Exosome-Mediated NeuroprotectionWu, Xie, Cao
et alJ Ethnopharmacol (2025)
Abstract: Post-stroke depression (PSD) is common among stroke survivors and negatively impacts recovery. Chaihu-Shugan-San (CSS), a traditional Chinese medicine, has shown therapeutic potential for mood disorders, particularly PSD. Recent studies suggest that CSS's effects may be mediated by exosomes, but the mechanisms remain unclear.This study aimed to evaluate the therapeutic effects of CSS on PSD in mice and investigate the underlying mechanisms, particularly the role of exosomes.Active compounds in CSS were identified from rat serum using liquid chromatography-mass spectrometry (LC-MS) and analyzed through network pharmacology. In vitro, an oxygen-glucose deprivation/reperfusion (OGD/R) BV2 microglia model was used to assess the effects of CSS-containing serum (CSS-S). Exosomes from OGD/R-treated BV2 microglia were isolated, labeled with PKH26, and analyzed using transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). In vivo, a photothrombotic stroke (PT) model combined with chronic unpredictable mild stress (CUMS) was used to induce PSD in mice. Behavioral assessments and histological analysis were performed, along with immunofluorescence (IF), ELISA and q-PCR to measure key protein and miR-146 expression in the hippocampus.CSS treatment significantly alleviated depressive-like behaviors in the PSD mouse model. Mice treated with high-dose CSS (4.2 g/kg) exhibited increased sucrose preference, reduced immobility in the tail suspension test (TST) and forced swimming test (FST), and enhanced exploratory activity in the open field test (OFT). Histological analysis demonstrated that CSS treatment improved brain tissue integrity, alleviating neuronal damage and reducing neuroinflammation. Exosome analysis revealed that CSS increased the expression of microglia-derived exosomes in the hippocampus, which were shown to carry miR-146. Further examination of miR-146 isoforms in the hippocampal tissue revealed significant changes: miR-146b-3p and miR-146a-5p were upregulated, while miR-146a-3p and miR-146b-5p were downregulated in PSD mice. Treatment with CSS reversed the altered miRNA expression, indicating a potential mechanism for its neuroprotective effects. Additionally, CSS treatment reduced the expression of inflammatory cytokines such as S100A8, IL1β, IL6, and TNF-α, while restoring the levels of angiogenic factors VEGFC and VEGFR3. ELISA measurements showed significant decreases in cyclic AMP response element-binding protein (CREB), brain-derived neurotrophic factor (BDNF), 5-hydroxytryptamine (5-HT), dopamine (DA), and noradrenaline (NE) in PSD mice; high-dose CSS notably elevated CREB and BDNF levels and showed comparable effects to fluoxetine in restoring 5-HT and DA levels. Additionally, the calcium signaling pathway was implicated, with altered mRNA expressions of CaMKIIα, CREB, phosphorylated CREB (p-CREB), PDE4D, and BDNF, although fluoxetine demonstrated stronger modulatory effects than CSS.CSS alleviates PSD in mice by modulating exosome-mediated signaling, particularly through the regulation of miR-146. The treatment reversed abnormal miRNA expression, reduced neuroinflammation, and improved synaptic function. These findings highlight CSS's potential as an effective therapeutic strategy for PSD by targeting exosome-mediated neuroprotection and miR-146 regulation.Copyright © 2025. Published by Elsevier B.V.
Deciphering the preeclampsia-specific immune microenvironment and the role of pro-inflammatory macrophages at the maternal-fetal interfaceFei, Lu, Shi
et alElife (2025) 13
Abstract: Preeclampsia (PE), a major cause of maternal and perinatal mortality with highly heterogeneous causes and symptoms, is usually complicated by gestational diabetes mellitus (GDM). However, a comprehensive understanding of the immune microenvironment in the placenta of PE and the differences between PE and GDM is still lacking. In this study, cytometry by time of flight indicated that the frequencies of memory-like Th17 cells (CD45RA-CCR7+IL-17A+CD4+), memory-like CD8+ T cells (CD38+CXCR3-CCR7+Helios-CD127-CD8+) and pro-inflam Macs (CD206-CD163-CD38midCD107alowCD86midHLA-DRmidCD14+) were increased, while the frequencies of anti-inflam Macs (CD206+CD163-CD86midCD33+HLA-DR+CD14+) and granulocyte myeloid-derived suppressor cells (gMDSCs, CD11b+CD15hiHLA-DRlow) were decreased in the placenta of PE compared with that of normal pregnancy (NP), but not in that of GDM or GDM&PE. The pro-inflam Macs were positively correlated with memory-like Th17 cells and memory-like CD8+ T cells but negatively correlated with gMDSCs. Single-cell RNA sequencing revealed that transferring the F4/80+CD206- pro-inflam Macs with a Folr2+Ccl7+Ccl8+C1qa+C1qb+C1qc+ phenotype from the uterus of PE mice to normal pregnant mice induced the production of memory-like IL-17a+Rora+Il1r1+TNF+Cxcr6+S100a4+CD44+ Th17 cells via IGF1-IGF1R, which contributed to the development and recurrence of PE. Pro-inflam Macs also induced the production of memory-like CD8+ T cells but inhibited the production of Ly6g+S100a8+S100a9+Retnlg+Wfdc21+ gMDSCs at the maternal-fetal interface, leading to PE-like symptoms in mice. In conclusion, this study revealed the PE-specific immune cell network, which was regulated by pro-inflam Macs, providing new ideas about the pathogenesis of PE.© 2024, Fei, Lu, Shi et al.
Calprotectin-mediated survival of Staphylococcus aureus in coculture with Pseudomonas aeruginosa occurs without nutrient metal sequestrationLee, Zygiel, Lee
et almBio (2025)
Abstract: Pseudomonas aeruginosa and Staphylococcus aureus are bacterial pathogens of major clinical concern that cause polymicrobial infections in diverse patient populations. Human calprotectin (CP; S100A8/S100A9 heterooligomer, MRP8/MRP14 heterooligomer) is a host-defense protein that contributes to nutritional immunity by sequestering multiple nutrient metal ions including Mn(II), Fe(II), and Zn(II). Here, we examine the consequences of metal availability and CP treatment on cocultures of P. aeruginosa and S. aureus. We report that CP elicits Fe-starvation responses in both P. aeruginosa and S. aureus in coculture, including the upregulation of genes involved in Fe uptake by both organisms. Moreover, analyses of pseudomonal metabolites in coculture supernatants further demonstrate Fe-starvation responses, showing that CP treatment leads to increased siderophore levels and reduced phenazine levels. Consistent with prior studies, growth under conditions of Fe depletion accelerated P. aeruginosa killing of S. aureus in coculture, but treatment with CP promoted S. aureus survival. Treatment with CP site variants lacking functional transition-metal-binding sites and metalated CP also enhanced S. aureus survival in coculture with P. aeruginosa, revealing that this consequence of CP treatment is independent of its canonical metal-sequestering function. Thus, the protective effects of CP treatment during coculture appear to override the observed Fe-starvation effects that make P. aeruginosa more virulent toward S. aureus. This work highlights an unappreciated facet of how CP contributes to host-pathogen and pathogen-pathogen interactions that are relevant to human infectious disease.The current working model that describes how the innate immune protein calprotectin (CP) protects the host against bacterial pathogens focuses on its capacity to sequester multiple essential metal nutrients in a process called nutritional immunity. Our study further explores this function by focusing on the effects of metal availability and CP treatment on the dynamics of Pseudomonas aeruginosa and Staphylococcus aureus grown in coculture. These two bacterial pathogens are of significant clinical concern and colocalize with CP at infection sites. This work reveals that CP modulates P. aeruginosa/S. aureus coculture dynamics in a manner that is independent of its ability to sequester nutrient metal ions. This surprising result is important because it demonstrates that CP has metal-independent function and thus contributes to the host-pathogen and pathogen-pathogen interactions in ways that are not accounted for in the current working model focused on metal sequestration.
New insights into tumor microenvironment and HPV integrations in cervical cancer pathogenesis revealed by single-cell transcriptome dataZeng, Peng, Wang
et alHum Mol Genet (2025)
Abstract: HPV infection is common among women and can result in serious illnesses. This research utilizes single-cell RNA-sequencing (scRNA-seq) to study the connection between cellular heterogeneity and HPV integrations in cervical histopathology. scRNA-seq was used to examine heterogeneity among normal patients and those in three disease stages: high-grade squamous intraepithelial lesions (HSIL), microinvasive carcinoma (MIC), and cervical squamous epithelium carcinoma cancer (CSCC) tissues. A method was developed to identify HPV integration events from scRNA-seq data. Our results indicated an increase in squamous epithelial cells and a decrease in columnar epithelial cells as the disease progressed from normal to CSCC. We discovered HPV genes that were differentially expressed across normal patients and those in the three disease stages. Notably, HPV integration events were more common in squamous epithelial cells at the single-cell level. The ratio of HPV-integrated cells increased as the disease progressed from normal tissue to CSCC, eventually stabilizing. Several genes, such as EGR1, S100A11, S100A8, KRT5, RPL34, ATP1B1, RPS4X and EEF2, were frequently integrated by HPV across patients. In contrast, genes like PAN3, BABAM2, SPEN, TCIM-SIRLNT, TEX41-PABPC1P2 and KCNV1-LINC01608 showed frequent integration events across cells. KRT5, ATP1B1, RPS4X, PAN3 and SPEN were novel recurrent HPV-integrated genes we observed at the patient or cell level in this study. Additionally, we found that HPV genes from various HPV types exhibited integration preferences in various samples and disease stages. This provides a valuable insight into the mechanism of HPV-induced cervical cancer from a single-cell standpoint, highlighting its clinical relevance.© The Author(s) 2025. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.
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