基于转录组分析三七总皂苷缓解阿司匹林所致小肠损伤的机制  

Mechanism of Panax notoginseng saponins on alleviating Aspirin-induced intestinal injury based on transcriptome analysis

作　　者：高若瑜 华国栋 朱宝琛 刘文卉 黄鑫 王萌 薛春苗[2] GAO RuoYu;HUA Guodong;ZHU Baochen(Beijing University of Chinese Medicine,Beijing 102488,China;Dongzhimen Hospital,Beijing University of Traditional Chinese Medicine,Beijing 100700,China)

机构地区：[1]北京中医药大学中药学院,北京102488 [2]北京中医药大学东直门医院药学部,北京100700

出　　处：《四川中医》2024年第10期57-64,共8页Journal of Sichuan of Traditional Chinese Medicine

基　　金：国家自然科学基金青年项目(编号:82004125);国家中医药管理局高水平重点学科临床中药学(编号:zyyzdxk-2023257)。

摘　　要：目的:研究三七总皂苷(PNS)缓解阿司匹林(ASA)所致大鼠小肠隐窝上皮细胞(IEC-6)损伤的潜在保护作用机制。方法:利用转录组学来阐明对照组、阿司匹林组和阿司匹林+三七总皂苷组IEC-6细胞中独特表达的基因,并利用GO和KEGG对差异表达基因进行富集分析。基于转录组学结果,建立阿司匹林诱导的小肠隐窝上皮细胞损伤的细胞模型,采用三七总皂苷进行干预,qRT-PCR法检测焦亡相关基因NLRP3、Casepase-1、IL-1βmRNA的表达量,CCK8法检测细胞生长率,Western blot检测细胞NLRP3、Casepase-1、IL-1β蛋白表达,探讨三七总皂苷通过抑制细胞焦亡减轻阿司匹林致小肠损伤的机制。结果:转录组测序分析显示,对照组与阿司匹林组共有6077个基因表达差异,而阿司匹林组与阿司匹林+三七总皂苷组有329个基因表达差异。GO功能富集揭示了多种关键生物过程、细胞成分、分子功能和通路进程,KEGG显著差异通路集中在炎性通路及细胞死亡(凋亡、焦亡)相关通路,提示阿司匹林对细胞的死亡方式及炎症反应有较强的干预作用。在体外实验中,阿司匹林能有效抑制IEC-6细胞的增殖,增加NLRP3、Casepase-1、IL-1βmRNA的表达量,促进NLRP3、Casepase-1、IL-1β蛋白的表达,促进细胞焦亡。与阿司匹林组相比,三七总皂苷和阿司匹林联合治疗可显著提高细胞增殖率,使NLRP3、Casepase-1、IL-1βmRNA的表达量降低,并抑制焦亡相关蛋白NLRP3、Casepase-1、IL-1β的表达。结论:我们的研究结果表明,三七总皂苷通过降低NLRP3、Casepase-1、IL-1βmRNA和蛋白的表达水平来抑制阿司匹林造成的小肠细胞焦亡,提示三七总皂苷在改善阿司匹林诱导的小肠损伤方面存在良好的潜力。Objective:To investigate the potential protective mechanism of Panax notoginseng saponins in alleviating aspirin-induced injury to small intestinal crypt epithelial cells in rats.Methods:Transcriptome analysis was employed to elucidate the uniquely expressed genes in IEC-6 cells of the control group,aspirin group,and aspirin+Panax notoginseng saponins group.Enrichment analysis of differentially expressed genes was conducted using GO and KEGG.Based on the transcriptome results,a cell model of small intestinal crypt epithelial cell injury induced by aspirin(ASA)was established.Pannotoginseng saponins(PNS)were used for intervention,qRT-PCR was utilized to detect the mRNA expression of pyroptosis-related genes NLRP3,Caspase-1,and IL-1β.The CCK8 method was applied to measure the cell growth rate.Western blotting was performed to examine the protein expressions of NLRP3,Caspase-1,and IL-1βin cells with an aim to explore the mechanism through which Panax notoginseng saponins alleviate aspirin-induced small intestinal injury by inhibiting pyroptosis.Results:Transcriptome sequencing analysis revealed 6077 differentially expressed genes between the control group and the aspirin group while 329 differentially expressed genes between the aspirin group and the combination treatment with panax notoginseng saponins group.GO functional enrichment unveiled various key biological processes,cellular components,molecular functions as well as pathway processes.KEGG significantly distinct pathways were concentrated in inflammatory pathways and cell death(apoptosis and pyroptosis)-related pathways indicating that aspirin had a strong intervention effect on cell death and inflammatory response.In vitro experiments demonstrated that aspirin effectively inhibited IEC-6 cell proliferation while increasing mRNA expression levels of NLRP3,Caspase-1,and IL-1β;it also promoted protein expression levels of NLRP3,Caspase-1,and IL-1βthereby promoting pyroptosis.Compared with the aspirin group,the combined treatment of panax notoginseng saponi

关 键 词：阿司匹林 三七总皂苷 小肠损伤 转录组测序 细胞焦亡 

分 类 号：R285.5[医药卫生—中药学]