【摘要】 背景与目的：研究发现，肝内免疫调节与肝纤维化发生发展密切相关。然而，在肝纤维化中与免疫细胞相关联的关键基因目前仍不清楚。因此，本研究探讨肝纤维化中的关键基因与疾病进展以及肝内免疫细胞分布的关联。方法：基于公共数据库中的不同肝纤维化程度的肝组织（GSE162694和GSE49541），对转录组测序RNA-seq测序数据进行差异表达比较，以正常肝样本作为对照；通过相对表达丰度排秩（REO）算法结果获得相关逆转稳定基因对；分析与肝纤维化相关的关键基因。构建CCl4诱导的肝纤维化小鼠模型，Masson染色鉴定病理组织学改变，qRT-PCR和Western blot方法检测关键基因的mRNA与蛋白表达；xCell工具分析关键基因与免疫细胞、肝纤维化进展的关系。结果：分析测序数据获得两个肝纤维化相关的交叠的逆转基因对（THBS2>RHDE及LBH>LRRC19）；其中THBS2、LBH均在肝纤维化组织中差异上调，并且LBH表达与肝纤维化分级、组织学评分和炎症评分明显相关（均P<0.05）。与对照组比较，模型组小鼠肝脏中LBH mRNA及蛋白水平均明显上调（均P<0.05）。基于LBH表达中位值，将肝纤维化芯片GSE162694、GSE49541中的样本分为LBH高表达组与LBH低表达组，xCell分析显示，CD⁴⁺记忆T细胞、中央记忆CD⁸⁺T细胞、树突状细胞、aDC、cDC等免疫细胞富集水平及免疫细胞评分在LBH高表达组中均明显上调（均P<0.05）。结论：转录辅助因子LBH可能参与调节肝内免疫细胞分布并促进肝纤维化的进展。更多还原

【Abstract】 Background and Aims: Previous studies have found a close association between intrahepatic immune regulation and the development of liver fibrosis. However, the key genes associated with immune cells in liver fibrosis are still unclear. Therefore, this study aimed to investigate the correlation between key genes in liver fibrosis, disease progression, and the distribution of intrahepatic immune cells.Methods: Differential gene expression analysis was performed on RNA sequencing data（RNA-seq） from liver tissues with different degrees of liver fibrosis obtained from public databases（GSE162694 and GSE49541）, using normal liver samples as controls. The relative expression order（REO） algorithm was used to identify relative reversal stabile pairs. Key genes associated with liver fibrosis were analyzed. A CCl4-induced mouse model of liver fibrosis was established, and histopathological changes were identified using Masson staining. The mRNA and protein expression of key genes were detected using qRT-PCR and Western blot methods, respectively. The relationship between key genes and immune cells as well as the progression of liver fibrosis was analyzed using the xCell tool.Results: Analysis of the sequencing data identified two overlapping reverse gene pairs associated with liver fibrosis（THBS2>RHDE and LBH>LRRC19）. Both THBS2 and LBH were upregulated in liver fibrosis tissues, and the expression of LBH was significantly correlated with fibrosis stage, histological score, and inflammation score（all P<0.05）. Compared to the control group, the mRNA and protein levels of LBH in the liver of the model mice were significantly upregulated（both P<0.05）. Based on the median expression value of LBH, the samples from the liver fibrosis datasets GSE162694 and GSE49541 were divided into the LBH high-expression group and the LBH low-expression group. The xCell analysis revealed that CD⁴⁺ memory T cells, central memory CD⁸⁺ T cells, dendritic cells, aDC, cDC, and other immune cells were enriched and immune cell scores were significantly upregulated in the high LBH expression group（all P<0.05）.Conclusion: The transcriptional co-factor LBH may regulate the distribution of intrahepatic immune cells and promote the progression of liver fibrosis.更多还原