基于转录组测序分析芹菜素抑制巨噬细胞焦亡的潜在作用机制  

The potential mechanism of apigenin inhibits macrophage pyroptosis based on transcriptome sequencing

作　　者：白勒谢 吴帅凯 叶利经 潘玉芝 吴丹媚 赵国军 BAI Lexie;WU Shuaikai;YE Lijing;PAN Yuzhi;WU Danmei;ZHAO Guojun(College of Pharmacy,Dali University,Dali,Yunnan 671000,China;Affiliated Qingyuan Hospital,Guangzhou Medical University/Qingyuan People′s Hospital,Qingyuan,Guangdong 511518,China;School of Intelligent Medicine and Biotechnology,Guilin Medical University,Guilin,Guangxi 541199,China)

机构地区：[1]大理大学药学院,云南大理671000 [2]广州医科大学附属清远医院/清远市人民医院,广东清远511518 [3]桂林医学院智能医学与生物技术学院,广西桂林541199

出　　处：《现代医药卫生》2025年第4期828-835,共8页Journal of Modern Medicine & Health

基　　金：国家自然科学基金项目(81870337);广东省自然科学基金面上项目(2024A1515012850)。

摘　　要：目的探讨芹菜素对巨噬细胞焦亡的影响及机制。方法体外培养THP-1细胞,应用佛波酯将其诱导分化为巨噬细胞,以脂多糖(LPS)和尼日利亚菌素联合刺激诱导巨噬细胞焦亡。通过光学显微镜观察细胞形态,采用Western blot检测Gasdermin D-N(GSDMD-N)蛋白的表达,应用实时荧光定量聚合酶链反应(qRT-PCR)检测NLRP3、IL-1β和TNF-αmRNA的表达,探讨芹菜素对巨噬细胞焦亡的影响。进一步使用转录组测序方法筛选差异表达基因和信号通路。最后应用qRT-PCR验证5.8S rRNA、18S rRNA、28S rRNA、Pre-45S rRNA及POP1、NAT10、RCL1和GAR1相关基因的表达,探讨核糖体生物合成信号通路在芹菜素抑制巨噬细胞焦亡中的作用。结果芹菜素预处理能够抑制巨噬细胞焦亡中气泡状突出物的形成,减少GSDMD-N蛋白和NLRP3、IL-1β、TNF-αmRNA的表达。基因本体(GO)富集分析结果显示芹菜素主要参与核酸代谢过程、RNA代谢过程和转录调节活性等功能,京都基因与基因组百科全书(KEGG)富集分析结果显示芹菜素主要参与核糖体生物合成、NOD样受体和Toll样受体等细胞焦亡相关信号通路。qRT-PCR验证显示,芹菜素预处理能够显著减少5.8S rRNA、18S rRNA、28S rRNA、Pre-45S rRNA及RCL1、GAR1、POP1和NAT10相关基因的表达。结论芹菜素对巨噬细胞焦亡具有抑制作用,其机制可能与抑制核糖体生物合成信号通路有关。Objective To explore the effect and mechanism of apigenin on macrophage pyroptosis.Methods THP-1 cells were cultured in vitro and induced to differentiate into macrophages by phorbol ester,and the macrophages were induced to pyroptosis by lipopolysaccharide(LPS)and nigericin.The cell morphology was observed by optical microscope.Western blot was used to detect the Gasdermin D-N(GSDMD-N).The mRNA expressions of NLRP3,IL-1βand TNF-αwere detected by real-time fluorescent quantitative polymerase chain reaction(qRT-PCR)to investigate the effect of apigenin on macrophage pyroptosis.Transcriptome sequencing was further used to screen differentially expressed genes and signaling pathways.Finally,qRT-PCR was used to verify the expression of 5.8S rRNA,18S rRNA,28S rRNA,Pre-45S rRNA and POP1,NAT10,RCL1 and GAR1 related genes,and to explore the role of ribosome biosynthesis signaling pathway in apigenin inhibiting macrophage pyroptosis.Results Apigenin pretreatment inhibited the formation of bubble-like protrusions during macrophage pyroptosis,and reduced the expression of GSDMD-N protein and the mRNA expression of NLRP3,IL-1β,and TNF-α.Gene ontology(GO)enrichment analysis showed that apigenin was mainly involved in nucleic acid metabolic process,RNA metabolic process and transcription regulator activity.Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis showed that apigenin mainly participates in ribosome biogenesis,NOD-like receptor signaling pathway and Toll-like receptor signaling pathway related to pyroptosis.qRT-PCR showed that apigenin pretreatment significantly reduced the expression of 5.8S rRNA,18S rRNA,28S rRNA,Pre-45S rRNA,RCL1,GAR1,POP1 and NAT10.Conclusion Apigenin has an inhibitory effect on macrophage pyroptosis,and its mechanism may be related to the regulation of the ribosome biogenesis signaling pathway.

关 键 词：巨噬细胞 细胞焦亡 芹菜素 转录组测序 核糖体生物合成信号通路 

分 类 号：R285.5[医药卫生—中药学]