PHYTOMEDICINE
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2025
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IF 8.3
Xue Yang,Jinni Yang,Anhui Wang,Yuan Zheng,Jiachun Lin,Ziwen Kong,Yuan Tian,Haijuan Dong,Zunjian Zhang,Rui Song
Background : Ulcerative colitis (UC) has demonstrated an escalating global incidence and prevalence, thereby posing substantial challenges to public health. Despite recent advancements in therapeutic interventions, the clinical management of UC remains suboptimal, underscoring the urgent need for novel treatment strategies. Saikosaponin A (SSa), a bioactive compound derived from the traditional Chinese herb Radix Bupleuri (RB), exhibits potent anti-inflammatory and immunomodulatory effects. However, its effects on UC and the underlying molecular mechanisms remain to be thoroughly explored. Purpose : This study aims to elucidate the underlying mechanisms of SSa in ameliorating UC and establish a pharmacological foundation for developing novel treatment modalities to address unmet clinical needs in UC treatment. Methods : The protective effects of SSa against DSS-induced acute colitis were evaluated in a 3% DSS-treated mouse model. Histological (H&E staining) and molecular analyses (RT-qPCR, ELISA, Western blotting, and flow cytometry) were performed to assess colonic tissue damage and inflammatory responses. Macrophage depletion via tail vein injection of clodronate liposomes confirmed the pivotal role of macrophages in UC pathogenesis and SSa's anti-inflammatory effects. The inhibitory effects of SSa on NLRP3 inflammasome activation were analyzed in vivo and in LPS/ATP-stimulated bone marrow-derived macrophages (BMDMs) using RT-qPCR, ELISA, and Western blotting. Bioinformatics analysis, targeted LC-MS/MS, and molecular docking were employed to identify potential molecular targets and mechanisms of SSa. Drug affinity responsive target stability (DARTS), cellular thermal shift assay (CETSA), and CH25H siRNA knockdown assays were used to validate CH25H as the direct target of SSa. Results : SSa effectively attenuated DSS-induced colitis in mice by alleviating colonic inflammation, preserving intestinal barrier integrity, reducing LPS translocation, and mitigating systemic organ injury in the liver and spleen. The inflammatory response of macrophages and the production of IL-1β were identified as key pathogenic components in colitis, and the clearance of macrophages significantly ameliorated colitis progression while SSa administration post-macrophage clearance did not further alter the disease phenotype. Mechanistically, SSa inhibited the NLRP3 inflammasome activation-mediated IL-1β secretion in macrophages. The sterol metabolism played a crucial role in the activation of the NLRP3 inflammasome. SSa also restored the disturbed sterol homeostasis in the colon under inflammatory conditions, especially promoted the synthesis of 25-hydroxycholesterol (25-OHC). Further investigation revealed that SSa primarily exerts its therapeutic effects by directly targeting cholesterol 25-hydroxylase (CH25H), which promotes the production of 25-OHC and inhibits macrophage NLRP3 inflammasome activation. Conclusion : This pioneering study demonstrated the therapeutic effect of SSa on DSS-induced colitis by targeting CH25H to enhance 25-OHC biosynthesis, which subsequently inhibited NLRP3 inflammasome activation in IL-1β-producing macrophages. These findings reveal a novel mechanism of SSa in UC treatment through cholesterol metabolism-regulated cascade immune modulation, providing strong pharmacological support for its development as a potential UC therapy.
研究领域:
免疫学
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药理学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
INFLAMMATION RESEARCH
·
2023
·Vol. 72
·期 5
·1099-1119
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IF 6.7
Ding Xuexuan,Qin Jingtong,Huang Fangfang,Feng Fuhai,Luo Lianxiang
Background Lipids, significant signaling molecules, regulate a multitude of cellular responses and biological pathways in asthma which are closely associated with disease onset and progression. However, the characteristic lipid genes and metabolites in asthma remain to be explored. It is also necessary to further investigate the role of lipid molecules in asthma based on high-throughput data. Objective To explore the biomarkers and molecular mechanisms associated with lipid metabolism in asthma. Methods In this study, we selected three mouse-derived datasets and one human dataset (GSE41665, GSE41667, GSE3184 and GSE67472) from the GEO database. Five machine learning algorithms, LASSO, SVM-RFE, Boruta, XGBoost and RF, were used to identify core gene. Additionally, we used non-negative matrix breakdown (NMF) clustering to identify two lipid molecular subgroups and constructed a lipid metabolism score by principal component analysis (PCA) to differentiate the subtypes. Finally, Western blot confirmed the altered expression levels of core genes in OVA (ovalbumin) and HDM+LPS (house dust mite+lipopolysaccharide) stimulated and challenged BALB/c mice, respectively. Results of non-targeted metabolomics revealed multiple differentially expressed metabolites in the plasma of OVA-induced asthmatic mice. Results Cholesterol 25-hydroxylase (CH25H) was finally localized as a core lipid metabolism gene in asthma and was verified to be highly expressed in two mouse models of asthma. Five-gene lipid metabolism constructed from CYP2E1, CH25H, PTGES, ALOX15 and ME1 was able to distinguish the subtypes effectively. The results of non-targeted metabolomics showed that most of the aberrantly expressed metabolites in the plasma of asthmatic mice were lipids, such as LPC 16:0, LPC 18:1 and LPA 18:1. Conclusion Our findings imply that the lipid-related gene CH25H may be a useful biomarker in the diagnosis of asthma.
研究领域:
生物信息学
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分子生物学
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免疫学
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药理学
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产品关联
DOI
PubMed
高影响因子
IMMUNITY
·
2024
·
IF 25.5
Jun Xiao,Shuang Wang,Longlong Chen,Xinyu Ding,Yuanhao Dang,Mingshun Han,Yuxiao Zheng,Huan Shen,Sifan Wu,Mingchang Wang,Dan Yang,Na Li,Chen Dong,Miao Hu,Chen Su,Weiyun Li,Lijian Hui,Youqiong Ye,Huiru Tang,Bin Wei,Hongyan Wang
Macrophages are critical to turn noninflamed “cold tumors” into inflamed “hot tumors”. Emerging evidence indicates abnormal cholesterol metabolites in the tumor microenvironment (TME) with unclear function. Here, we uncovered the inducible expression of cholesterol-25-hydroxylase (Ch25h) by interleukin-4 (IL-4) and interleukin-13 (IL-13) via the transcription factor STAT6, causing 25-hydroxycholesterol (25HC) accumulation. scRNA-seq analysis confirmed that CH25H hi subsets were enriched in immunosuppressive macrophage subsets and correlated to lower survival rates in pan-cancers. Targeting CH25H abrogated macrophage immunosuppressive function to enhance infiltrating T cell numbers and activation, which synergized with anti-PD-1 to improve anti-tumor efficacy. Mechanically, lysosome-accumulated 25HC competed with cholesterol for GPR155 binding to inhibit the kinase mTORC1, leading to AMPKα activation and metabolic reprogramming. AMPKα also phosphorylated STAT6 Ser564 to enhance STAT6 activation and ARG1 production. Together, we propose CH25H as an immunometabolic checkpoint, which manipulates macrophage fate to reshape CD8 + T cell surveillance and anti-tumor response.
研究领域:
免疫学
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肿瘤学
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代谢生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
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