类风湿关节炎相关性间质性肺病(RA-ILD)是RA患者常见的并发症之一,影响他们的生活质量。CIBERSORT算法广泛用于确定患病组织中免疫细胞(IC)的比例,而SonicHedgehog(Shh)信号通路,作为一个必要的调节因素,在RA-ILD的病理学方面也引起了关注。本工作旨在基于CIBERSORT算法探索RA-ILD免疫浸润和滑膜组织(ST)Shh表达的机制。使用R语言对RA-ILD的差异基因进行途径富集分析。使用基于机器学习的CIBERSORT算法分析RA-ILD肺组织中22种IC的含量和比例。同时,免疫印迹法检测和分析Shh的表达,平滑(Smo),RA-ILD和Ctrl组(无ILD的RA患者)的ST样品中的骨形态发生蛋白(BMP)蛋白。与RA-ILD相关的蛋白质网络中的枢纽靶基因包括BSG,CCL2,CTLA4,FGFBP1,GLI1,HHIP,HLA-DRB1,IFNAR1,IL17A,IL23a,IL-6,INPP4A,LILRB1,MUC5B,PADI4,PPM1A,PTCH1,PTPN22,RSPO4,嘘,SMO,STAT4,SUFU,TAOK2、TIMP2和TWSG1参与多种途径,如B细胞调节,Shh通路的转录因子,和ST免疫耐受相关途径。在使用CIBERSORT算法对RA-ILD进行免疫学分析时,HLA(r=-0.26),PTPN22(r=-0.36),STAT4(r=-0.18),IL-6(r=-0.17),CTLA4(r=-0.27),和PADI4(r=-0.21)均与CD4T细胞呈负相关(P<0.05)。与Ctrl组相比,在RA-ILD患者血清中发现单核细胞更丰富。嘘,Smo,与Ctrl组相比,RA-ILD组BMP表达明显降低(P<0.05)。在RA-ILD患者的肺组织中观察到明显的免疫细胞浸润。利用CIBERSORT算法的进一步分析揭示了不同IC亚型比例的变化,表明它们与疾病严重程度和预后有关。此外,Shh的表达水平显着降低,Smo,和BMP。这些发现强调了免疫细胞在RA-ILD病理生理学中的重要性,并表明Shh信号通路可能参与RA-ILD的发病机理。
Rheumatoid arthritis-related interstitial lung disease (RA-ILD) is one of the common complications in patients with RA, which affects their quality of life. The CIBERSORT algorithm is widely employed to determine the proportion of immune cells (ICs) in diseased tissues, while the Sonic Hedgehog (Shh) signaling pathway, as an imperative regulatory factor, has also attracted attention in the pathology of RA-ILD. This work was to explore the mechanisms of RA-ILD immune infiltration and synovial tissue (ST) Shh expression based on the CIBERSORT algorithm. The differential genes of RA-ILD were subjected to pathway enrichment analysis using R language. The content and proportion of 22 types of ICs in RA-ILD lung tissues were analyzed using machine learning-based CIBERSORT algorithm. Meanwhile, immunoblotting was employed to detect and analyze the expression of Shh, Smoothened (Smo), and bone morphogenetic proteins (BMPs) proteins in ST samples from RA-ILD and Ctrl groups (RA patients without ILD). The hub target genes in the protein network associated with RA-ILD include BSG, CCL2, CTLA4, FGFBP1, GLI1, HHIP, HLA-DRB1, IFNAR1, IL17A, IL23A, IL-6, INPP4A, LILRB1, MUC5B, PADI4, PPM1A, PTCH1, PTPN22, RSPO4, Shh, SMO, STAT4, SUFU, TAOK2, TIMP2, and TWSG1, which are involved in multiple pathways, such as B cell regulation, transcription factors of the Shh pathway, and ST immune tolerance-related pathways. In the immunological analysis of RA-ILD using the CIBERSORT algorithm, HLA (r = - 0.26), PTPN22 (r = - 0.36), STAT4 (r = - 0.18), IL-6 (r = - 0.17), CTLA4 (r = - 0.27), and PADI4 (r = - 0.21) were all found to exhibit negative correlations with CD4+T cells (P < 0.05). Monocytes were found to be more abundant in RA-ILD patients\' serum versus the Ctrl group. Shh, Smo, and BMP expressions were drastically lower in the RA-ILD group versus Ctrl group (P < 0.05). Significant immune cell infiltration was observed in the lung tissues of RA-ILD patients. Further analysis utilizing the CIBERSORT algorithm revealed alterations in the proportions of different IC subtypes, indicating their association with disease severity and prognosis. Moreover, there was a significant decrease in the expression levels of Shh, Smo, and BMP. These findings underscore the importance of immune cells in the pathophysiology of RA-ILD and suggest a potential involvement of the Shh signaling pathway in the pathogenesis of RA-ILD.