Chakraborty Debolina, Joshi Lovely, Agnihotri Prachi, Malik Swati, Pal Niyati, Kumar Vijay, Biswas Sagarika
Council of Scientific & Industrial Research (CSIR)-Institute of Genomics and Integrative Biology, Mall Road, Delhi University Campus, Delhi, 110007, India.
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, India.
Mol Biol Rep. 2025 Apr 26;52(1):429. doi: 10.1007/s11033-025-10530-2.
Rheumatoid arthritis (RA) is a chronic autoimmune disease marked by joint damage and disrupted vitamin D signaling, leading to calcium deposition in joints. This study explores the therapeutic potential of Bavachin (BVN), a phytoestrogen from Psoralea corylifolia, in modulating vitamin D signaling in RA.
Vitamin D receptor (VDR) structure was modeled using SWISS-MODEL, AlphaFold, and I-TASSER, followed by docking with BVN and Estradiol (E2) via AutoDock Vina. BVN's effects on VDR mRNA and protein levels in RA-FLS were assessed by qRT-PCR and Western blot(WB). VDR-related BVN targets in RA were explored through protein-protein interaction (PPI) network and pathway analysis in Cytoscape. BVN's impact on RXRα expression and VDR-RXRα interaction was examined by WB and immunofluorescence. Alizarin staining evaluated calcium deposition, while qRT-PCR analyzed BVN's regulation of calcium-binding proteins.
In silico analysis revealed a strong interaction of VDR with BVN with Gibbs-free energy of -7.2 Kcal/mol with prominent H-bonds. Further, in vitro study in RA-FLS revealed that BVN treatment increased VDR mRNA and protein expression. PPI and pathway enrichment analysis retrieved RXRα as the prominent protein to be targeted by BVN in Vitamin D signaling. BVN treatment also significantly upregulated RXRα expression and enhanced the interaction between VDR and RXRα. Further, BVN modulated associated calcium signaling, reduced calcium deposition in RA-FLS and significantly downregulated calcium-binding proteins CALB1, CALB2, NCX1, TRPV5, and TRPV6.
Collectively, this study depicted a prominent therapeutic efficacy of BVN in targeting Vitamin D signaling and associated calcium deposition to alleviate RA pathogenesis.
类风湿性关节炎（RA）是一种慢性自身免疫性疾病，其特征为关节损伤和维生素D信号传导紊乱，导致钙在关节中沉积。本研究探讨了补骨脂中的一种植物雌激素补骨脂素（BVN）在调节RA中维生素D信号传导方面的治疗潜力。
使用SWISS-MODEL、AlphaFold和I-TASSER对维生素D受体（VDR）结构进行建模，然后通过AutoDock Vina将其与BVN和雌二醇（E2）进行对接。通过qRT-PCR和蛋白质印迹法（WB）评估BVN对RA - FLS中VDR mRNA和蛋白质水平的影响。通过Cytoscape中的蛋白质 - 蛋白质相互作用（PPI）网络和通路分析探索RA中与VDR相关的BVN靶点。通过WB和免疫荧光检查BVN对RXRα表达和VDR - RXRα相互作用的影响。茜素染色评估钙沉积，而qRT-PCR分析BVN对钙结合蛋白的调节作用。
计算机模拟分析显示VDR与BVN有很强的相互作用，吉布斯自由能为 -7.2千卡/摩尔，有显著的氢键。此外，对RA - FLS的体外研究表明，BVN处理可增加VDR mRNA和蛋白质表达。PPI和通路富集分析发现RXRα是BVN在维生素D信号传导中作用的主要靶点。BVN处理还显著上调RXRα表达，并增强VDR与RXRα之间的相互作用。此外，BVN调节相关钙信号传导，减少RA - FLS中的钙沉积，并显著下调钙结合蛋白CALB1、CALB2、NCX1、TRPV5和TRPV6。
总体而言，本研究表明BVN在靶向维生素D信号传导和相关钙沉积以减轻RA发病机制方面具有显著的治疗效果。
