| 引用本文: |
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[点击复制] |
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| 青藤碱通过调控XCL抑制Jurkat细胞增殖与活化的机制研究 |
| 徐丽,胡泽玉,胡明月,赵芳,蔡雄 |
| (湖南中医药大学中医药科学院, 湖南 长沙 410208;湖南中医药大学第一附属医院, 湖南 长沙 410007) |
| 摘要: |
| 目的 探究青藤碱对Jurkat细胞增殖与活化的影响及其通过调控XC基序趋化因子配体（XCL）表达在急性Ｔ淋巴细胞白血病中的潜在作用机制。方法 以人T淋巴细胞白血病细胞系（Jurkat, Clone E6-1）为研究对象，分别以不同浓度的青藤碱（0、150、300、600 μmol/L）处理24、48、72 h，通过CCK-8检测细胞增殖，以确定最佳干预浓度和时间；流式细胞术检测细胞凋亡；流式细胞术和ELISA检测T细胞活化标志物CD25表达水平及活化后白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、干扰素-γ（IFN-γ）的含量；转录组测序分析差异表达基因（DEGs）变化，并对DEGs进行GO和KEGG通路富集分析；比较各组间XCL的基因丰度，qRT-PCR验证细胞中XCL1和XCL2 mRNA的表达水平。结果 与青藤碱0 μmol/L组比较，青藤碱150 μmol/L组、青藤碱300 μmol/L组和青藤碱600 μmol/L组Jurkat细胞的存活率、T细胞活化标志物CD25表达水平、细胞上清液中IFN-γ含量均降低（P<0.05，P<0.01，P<0.001）；Jurkat细胞的凋亡率升高（P<0.01，P<0.001）。与青藤碱150 μmol/L组比较，青藤碱300 μmol/L组和青藤碱600 μmol/L组Jurkat细胞的凋亡率均升高（P<0.001）；细胞上清液中IFN-γ含量均降低（P<0.001）。与青藤碱300 μmol/L组比较，青藤碱600 μmol/L组Jurkat细胞的凋亡率升高（P<0.001）；细胞上清液中IFN-γ含量降低（P<0.001）。转录组测序结果发现，青藤碱0 μmol/L组、青藤碱300 μmol/L组和青藤碱600 μmol/L组共获得97个DEGs；GO富集分析表明，DEGs显著富集于免疫与炎症反应、对TNF的应答及TNF介导的信号通路等生物学过程，涉及的DEGs包括XCL2、XCL1等；KEGG通路富集分析进一步表明，XCL信号通路在青藤碱处理组中明显富集。基因表达水平分析显示，DEGs中XCL在青藤碱0 μmol/L组中高表达，而在青藤碱300 μmol/L组和青藤碱600 μmol/L组中低表达；进一步qRT-PCR结果证实，与青藤碱0 μmol/L组比较，青藤碱300 μmol/L组和青藤碱600 μmol/L组Jurkat细胞的XCL1和XCL2 mRNA表达水平均降低（P<0.001）。结论 青藤碱可抑制Jurkat细胞增殖与活化，其分子机制可能与XCL表达的下调相关。 |
| 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 |
| DOI：10.3969/j.issn.1674-070X.2025.12.007 |
| 投稿时间：2025-08-21 |
| 基金项目:国家自然科学基金面上项目（82274506）；湖南省自然科学基金面上项目（224JJ5298）。 |
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| The mechanism of sinomenine in inhibiting Jurkat cell proliferation and activation through regulation of XCL |
| XU Li, HU Zeyu, HU Mingyue, ZHAO Fang, CAI Xiong |
| (Academy of Chinese Medical Sciences, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| 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. |
| Key words: sinomenine Jurkat cell cell proliferation cell activation transcriptome sequencing XC motif chemokine ligand |
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