湖南中医药大学学报2025，Vol.45Issue(12)：2288-2295,8.DOI:10.3969/j.issn.1674-070X.2025.12.007
青藤碱通过调控XCL抑制Jurkat细胞增殖与活化的机制研究
The mechanism of sinomenine in inhibiting Jurkat cell proliferation and activation through regulation of XCL
摘要
Abstract
Objective To investigate the effects of sinomenine(SIN)on Jurkat cell proliferation and activation as well as its potential mechanism of action in acute T-cell lymphoblastic leukemia via regulation of XC motif chemokine ligand(XCL)expression.Methods The human T lymphocytic leukemia cell line Jurkat(Clone E6-1)was treated with different concentrations of SIN(0,150,300,600 μmol/L)for 24,48,and 72 hours.Cell proliferation was assessed using the CCK-8 assay to determine the optimal intervention concentration and duration.Apoptosis was measured by flow cytometry.Cell activation and post-activation cytokine levels(IL-6,TNF-α,and IFN-γ)were measured via flow cytometry and ELISA.Flow cytometry and ELISA were employed to measure the expression level of the T cell activation marker CD25,as well as the levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and interferon-γ(IFN-γ)after activation.Transcriptome sequencing was performed to analyze differentially expressed genes(DEGs),followed by GO and KEGG pathway enrichment analysis of the DEGs.Gene abundance of XCL was compared across groups.The mRNA expression levels of XCL1 and XCL2 in cells were verified by qRT PCR.Results Compared with the 0 μmol/L SIN group,the 150 μmol/L,300 μmol/L,and 600 μmol/L SIN groups showed decreased cell viability,lower expression of the T cell activation marker CD25,and reduced IFN-γ levels in the cell supernatant(P<0.05,P<0.01,P<0.001).The apoptosis rate of Jurkat cells increased in these groups(P<0.01,P<0.001).Compared with the 150 μmol/L SIN group,the 300 μmol/L and 600 μmol/L SIN groups exhibited an elevation in apoptosis rate(P<0.001)and a reduction in IFN-γ levels(P<0.001).Compared with the 300 μmol/L SIN group,the 600 μmol/L SIN group displayed increased apoptosis(P<0.001)and decreased IFN-γ secretion(P<0.001).Transcriptome sequencing identified 97 DEGs among the 0,300,and 600 μmol/L SIN groups.GO enrichment analysis indicated that these DEGs were significantly enriched in biological processes related to immune and inflammatory responses,response to TNF,and TNF mediated signaling pathways,involving DEGs such as XCL2 and XCL1.KEGG pathway analysis further revealed marked enrichment of the XCL signaling pathway in SIN treated groups.Expression analysis showed that XCL was highly expressed in the 0 μmol/L SIN group but downregulated in the 300 μmol/L and 600 μmol/L groups.Subsequent qRT-PCR verification confirmed that the mRNA expression levels of both XCL1 and XCL2 were lower in the 300 μmol/L and 600 μmol/L SIN groups compared with the 0 μmol/L SIN group(P<0.001).Conclusion SIN can inhibit the proliferation and activation of Jurkat cells,and its molecular mechanism may be associated with the downregulation of XCL expression.关键词
青藤碱/Jurkat细胞/细胞增殖/细胞活化/转录组测序/XC基序趋化因子配体Key words
sinomenine/Jurkat cell/cell proliferation/cell activation/transcriptome sequencing/XC motif chemokine ligand分类
医药卫生引用本文复制引用
徐丽,胡泽玉,胡明月,赵芳,蔡雄..青藤碱通过调控XCL抑制Jurkat细胞增殖与活化的机制研究[J].湖南中医药大学学报,2025,45(12):2288-2295,8.基金项目
国家自然科学基金面上项目(82274506) （82274506）
湖南省自然科学基金面上项目(224JJ5298). （224JJ5298）
