基于生物信息学探讨泄浊解毒方调控细胞自噬治疗溃疡性结肠炎的机制
Bioinformatics Reveals Mechanism of Xiezhuo Jiedu Precription in Treatment of Ulcerative Colitis by Regulating Autophagy
康欣 1孙超迪 2刘建平 1任杰 1杜明民 1赵源 1郎晓猛1
作者信息
- 1. 河北省中医院,石家庄 050011;河北省中西医结合胃肠病研究重点实验室,石家庄 050011;河北省浊毒证重点实验室,石家庄 050011
- 2. 河北省中西医结合胃肠病研究重点实验室,石家庄 050011;河北省浊毒证重点实验室,石家庄 050011;邯郸市中医院,河北邯郸 056001
- 折叠
摘要
目的:利用生物信息学与动物实验方法探讨泄浊解毒方调节细胞自噬治疗溃疡性结肠炎(UC)潜在机制.方法:从基因表达综合数据库(GEO)中获取UC患者结肠黏膜组织差异表达基因,与细胞自噬基因取交集,获取细胞自噬差异基因(DEARG),将DEARG分别导入Metascape和STRING数据库,进行基因本体/京都基因与基因组百科全书(GO/KEGG)富集分析、蛋白质-蛋白质相互作用(PPI)分析,最终得到15个关键DEARG,通过最小绝对收缩和选取算子(LASSO)回归和接收者操作特征曲线(ROC)曲线分析获取核心DEARG;运用CIBERSORT去卷积法对UC患者样本做免疫浸润分析,以及核心DEARG与免疫细胞的相关性分析.将C57BL/6J小鼠分为正常组和模型制备组,采用2.5％的葡聚糖硫酸钠自由饮方式,建立UC小鼠模型,按照随机数字表法分为泄浊解毒低、中、高剂量组及美沙拉嗪组,灌胃7 d,苏木素-伊红染色观察结肠黏膜形态,蛋白免疫印迹法检测结肠组织中胱天蛋白酶-1(Caspase-1)、组织蛋白酶B(CTSB)、C-C基序趋化因子2(CCL2)、趋化因子(CXC基序)受体4(CXCR4)、缺氧诱导因子-1α(HIF-1α)蛋白表达量,实时荧光定量聚合酶链式反应法检测结肠组织中Caspase-1、CCL2、CTSB、CXCR4、HIF-1α mRNA表达.结果:从GEO数据库筛选出数据集GSE87466,与细胞自噬基因取交集,运用PPI分析、LASSO回归和ROC曲线分析后得到核心DEARG(Caspase-1、CCL2、CTSB、CXCR4),免疫浸润分析结果显示NK细胞、M0型巨噬细胞、M1型巨噬细胞、树突细胞在UC患者结肠黏膜组织中表达具有显著差异,核心DEARG与此类免疫细胞同样存在显著相关性.与泄浊解毒方网络药理学结果取交集后发现HIF-1α信号通路可能是泄浊解毒方调控UC的关键信号通路.动物实验观察到泄浊解毒方可明显缓解UC小鼠结肠黏膜炎症性反应,与正常组比较,模型组Caspase-1、CCL2、CTSB、CXCR4、HIF-1α蛋白和mRNA的表达量显著升高,经泄浊解毒方及美沙拉嗪治疗后,各组小鼠结肠黏膜中Caspase-1、CCL2、CTSB、CXCR4、HIF-1α蛋白和mRNA的表达量均显著下调.结论:Caspase-1、CCL2、CTSB、CXCR4是与UC发病密切相关的细胞自噬基因,泄浊解毒方可下调细胞自噬核心基因缓解小鼠结肠黏膜炎症反应.
Abstract
Objective:To explore the potential mechanism of Xiezhuo Jiedu prescription in regulating autophagy in the treatment of ulcerative colitis(UC)by bioinformatics and animal experiments.Methods:The differentially expressed genes(DEGs)in the colonic mucosal tissue of UC patients was obtained from the Gene Expression Omnibus(GEO),and those overlapped with autophagy genes were obtained as the differentially expressed autophagy-related genes(DEARGs).DEARGs were imported into Metascape and STRING,respectively,for gene ontology/Kyoto Encyclopedia of Genes and Genomics(GO/KEGG)enrichment analysis and protein-protein interaction(PPI)analysis.Finally,15 key DEARGs were obtained.The core DEARGs were obtained by least absolute shrinkage and selection operator(LASSO)regression and receiver operating characteristic curve(ROC)analysis.The CIBERSORT deconvolution algorithm was used to analyze the immunoinfiltration of UC patients and the correlations between core DEARGs and immune cells.C57BL/6J mice were assigned into a normal group and a modeling group.The mouse model of UC was established by free drinking of 2.5％dextran sulfate sodium.The modeled mice were assigned into low-,medium-,and high-dose Xiezhuo Jiedu prescription and mesalazine groups according to the random number table method and administrated with corresponding agents by gavage for 7 days.The colonic mucosal morphology was observed by hematoxylin-eosin staining.The protein and mRNA levels of cysteinyl aspartate-specific proteinase 1(Caspase-1),cathepsin B(CTSB),C-C motif chemokine-2(CCL2),CXC motif receptor 4(CXCR4),and hypoxia-inducing factor-1α(HIF-1α)in the colon tissue were determined by Western blot and real-time fluorescence quantitative polymerase chain reaction,respectively.Results:The dataset GSE87466 was screened from GEO and interlaced with autophagy genes.After PPI analysis,LASSO regression,and ROC analysis,the core DEARGs(Caspase-1,CCL2,CTSB,and CXCR4)were obtained.The results of immunoinfiltration analysis showed that the counts of NK cells,M0 macrophages,M1 macrophages,and dendritic cells in the colonic mucosal tissue of UC patients had significant differences,and core DEARGs had significant correlations with these immune cells.This result,combined with the prediction results of network pharmacology,suggested that the HIF-1α signaling pathway may play a key role in the regulation of UC by Xiezhuo Jiedu prescription.The animal experiments showed that Xiezhuo Jiedu prescription significantly alleviated colonic mucosal inflammation in UC mice.Compared with the normal group,the model group showed up-regulated protein and mRNA levels of caspase-1,CCL2,CTSB,CXCR4,and HIF-1α,which were down-regulated after treatment with Xiezhuo Jiedu prescription or mesalazine.Conclusion:Caspase-1,CCL2,CTSB,and CXCR4 are autophagy genes that are closely related to the onset of UC.Xiezhuo Jiedu prescription can down-regulate the expression of core autophagy genes to alleviate the inflammation in the colonic mucosa of mice.
关键词
溃疡性结肠炎/泄浊解毒方/生物信息学/细胞自噬Key words
ulcerative colitis/Xiezhuo Jiedu prescription/bioinformatics/autophagy引用本文复制引用
出版年
2025
国家科技期刊平台
NSTL
维普
万方数据