iTRAQ蛋白质组学鉴定亲环蛋白为马氏珠母贝中环孢素A免疫调节的候选介质

Abstract

ITRAQ-based proteomics identified cyclophilin as a candidate mediator for immunomodulation in pearl oyster. • CsA injection reduced the serum IL-17 levels while enhancing the activities of SOD and CAT. • CsA treatment altered the expression of genes associated with HIF-1 and Notch signaling pathways. • CsA significantly inhibited LPS-induced hemocyte apoptosis. Transplantation surgery induces severe immune stress in pearl oysters Pinctada fucata martensii , yet effective immunomodulatory strategies remain limited. This study establishes a proteomics-guided strategy for immunomodulation by identifying cyclophilin as a candidate mediator through iTRAQ-based quantitative analysis and validating cyclosporin A (CsA) through serum immune indicator and transcriptome analysis. Serum iTRAQ analysis identified 1069 differentially expressed proteins following transplantation, among which cyclophilin exhibited significant up-regulation and was selected as a candidate target for therapeutic intervention. Dose-response assays determined 3.9 × 10 −3  mol/L as the optimal concentration, which significantly suppressed TNF-α levels under LPS stimulation ( P  < 0.05). At this concentration, CsA pre-treatment markedly decreased circulating IL-17 concentrations, while it enhanced SOD and CAT enzymatic activities ( P  < 0.05). Transcriptome analysis revealed that CsA treatment was associated with upregulation of HIF-1 and Notch signaling pathway-related genes, concomitant with alleviation of LPS-induced downregulation in DNA replication and cell cycle pathways. Furthermore, CsA significantly inhibited hemocyte apoptosis compared to the control group ( P  < 0.05). These findings demonstrate that iTRAQ-based proteomic screening enables rational target identification for immunomodulation, and CsA exerts protective effects associated with altered expression of HIF-1/Notch pathway-related genes and apoptosis inhibition. These findings provide a molecular basis for developing immunomodulato