基于单细胞测序与微阵列芯片数据，系统探索溃疡性结肠炎（UC）的潜在治疗靶点及分子亚型特征，为疾病分型与个体化治疗提供分子依据。

从GEO数据库获取UC相关单细胞测序数据集GSE182270，选取3例UC样本与3例正常样本进行质量控制与分析，识别并注释12个细胞亚群。基于各亚群差异表达基因（DEGs）并结合炎症因子基因集筛选关键基因。随后下载并整合转录组数据集GSE75214与GSE87473，共包含203例UC样本与32例正常样本。利用sva包去除批次效应，limma包进行标准化处理，构建统一的UC表达矩阵。通过一致性聚类识别UC分子亚型，并采用主成分分析（PCA）验证亚型间的差异。免疫组化验证关键基因在UC样本中的表达。连续变量比较采用配对t检验，非正态分布变量采用Wilcoxon符号秩检验，多组间比较分析采用单因素方差分析。

共筛选出17个在UC样本中上调的关键基因（P < 0.05），包括CCL2、CCL20、CD14、CD48、CD69、CXCL10、F3、GPR183、IL-1β、IL7R、CXCL8、LCK、NAMPT、NFKBIA、PDPN、PLAUR、TIMP1。PCA显示，基于关键基因的一致性聚类能够有效区分不同UC分子亚型。HLA家族基因（HLA-B、HLA-C、HLA-DMA、HLA-DMB、HLA-DPA1、HLA-DQB1、HLA-F等17个）及37个免疫检查点基因在亚型4中表达升高，而HHLA2在亚型3中高表达（P < 0.05）。上述差异表达提示不同亚型存在免疫微环境特征差异。免疫组化显示，与对照组比较，3个核心基因编码的蛋白IL-1β，TIMP1和HLA-F在UC组中表达升高。

整合单细胞测序与转录组数据揭示，HLA家族及免疫检查点基因的差异表达可反映UC的分子亚型特征。IL-1β，TIMP1和HLA-F等基因在UC中具有重要的致病与调控作用，可能作为潜在的分子靶点，为UC的精准分型与个体化治疗提供新的生物学依据。

To systematically explore potential therapeutic targets and molecular subtype characteristics of ulcerative colitis (UC) by integrating single-cell RNA sequencing (scRNA-seq) and microarray data, providing a molecular basis for disease subtyping and personalized treatment.

The scRNA-seq dataset GSE182270, including 3 UC samples and 3 normal samples, was obtained from the GEO database. After quality control, 12 cell subpopulations were identified and annotated. Key genes were screened based on differentially expressed genes (DEGs) from these subpopulations and an inflammation-related gene set. Subsequently, transcriptomic datasets GSE75214 and GSE87473, including 203 UC samples and 32 normal samples, were integrated. Batch effects were corrected using the sva package, and data were normalized using the limma package to construct a unified UC expression matrix. Molecular subtypes were identified via consensus clustering, with differences validated by principal component analysis (PCA). Immunohistochemistry (IHC) was employed to examine the expression of key genes in UC samples. Continuous variables were compared using paired t-tests, non-normally distributed variables were analyzed using Wilcoxon signed-rank tests, and differences among multiple groups were analyzed using one-way analysis of variance.

Seventeen key genes (CCL2, CCL20, CD14, CD48, CD69, CXCL10, F3, GPR183, IL-1β, IL7R, CXCL8, LCK, NAMPT, NFKBIA, PDPN, PLAUR, TIMP1) were significantly upregulated in UC samples (P < 0.05). PCA confirmed that consensus clustering based on these key genes effectively distinguished distinct UC molecular subtypes. Seventeen HLA family genes (including HLA-B, HLA-C, HLA-DMA, HLA-DMB, HLA-DPA1, HLA-DQB1) and 37 immune checkpoint genes was significantly overexpression in subtype 4, while HHLA2 was highly expressed in subtype 3 (P < 0.05), indicating distinct immune microenvironment features across subtypes. Elevated expression of the proteins (IL-1β, TIMP-1, and HLA-F) encoded by three hub genes was observed in the UC group compared to controls by immunohistochemistry.

The integration of scRNA-seq and transcriptomic data reveals that differential expression of HLA family and immune checkpoint genes defines molecular subtypes of UC. Key genes such as IL-1β, TIMP1 and HLA-F play significant pathogenic and regulatory roles and represent potential molecular targets, offering new biological insights for the precise subtyping and personalized treatment of UC.