Heliyon (Nov 2024)
hsa_circ_0007755 competitively adsorbs miR-27b-3p to mediate CXCL2 expression and recruit Th1 cells to promote hypertrophic scars development
Abstract
Background and objective: The circular RNA hsa_circ_0007755 is markedly upregulated in hypertrophic scars (HS), yet its functional roles in this fibroproliferative disorder remain to be elucidated. This investigation aims to delineate the regulatory mechanisms of hsa_circ_0007755 in HS and to decode its downstream molecular signaling pathways. Methods: We established a murine model of HS. Tissue histopathology was assessed using Hematoxylin and Eosin and Masson's trichrome staining. Peripheral blood from the animals was collected and the ratio of T-helper 1 (Th1) to T-helper 2 (Th2) cells was quantified via flow cytometry. The proliferation and apoptosis rates of human hypertrophic scar fibroblasts (hHSFs) were evaluated using the Cell Counting Kit-8 assay and flow cytometry, respectively. The invasive capacity of hHSFs was assessed via a Transwell assay. Co-culture experiments of hHSFs with T cells were conducted, and alterations in Th1/Th2 ratios were monitored using flow cytometry. Levels of cytokines, fibrosis-associated proteins, nuclear factor-kappaB (NF-κB) pathway-related protein, and C-X-C Motif Chemokine Ligand 2 (CXCL2) were quantified using Enzyme-Linked Immunosorbent Assay or Western blot analysis. The interactions between hsa_circ_0007755, miR-27b-3p, and CXCL2 were investigated using dual-luciferase reporter assays and RNA immunoprecipitation. Results: Both hsa_circ_0007755 and CXCL2 were highly expressed in HS, whereas miR-27b-3p was downregulated. Knockdown of hsa_circ_0007755 inhibited the proliferation and invasion of hHSFs, promoted apoptosis, and reduced the expression of fibrotic proteins α-SMA and Collagen I, as well as the phosphorylation of the inflammatory pathway protein p65. Co-culture experiments confirmed that hHSFs lowly expressing hsa_circ_0007755 showed a decreased Th1 cell proportion and an increased Th2 cell proportion, alongside lower levels of TNF-α and INF-γ and higher levels of IL-4 and IL-10. The effects of either knocking down or overexpressing hsa_circ_0007755 were reversed by knocking down either miR-27b-3p or CXCL2, respectively. hsa_circ_0007755 acted as a ''molecular sponge'' for miR-27b-3p, sequestering and diminishing its availability, thereby alleviating its suppression of the target gene CXCL2. Conclusion: hsa_circ_0007755 plays a pivotal role in modulating the immune response of HS by influencing the miR-27b-3p/CXCL2 axis, regulating the function and proportion of Th1 and Th2 cells, and thereby affecting the inflammatory and fibrotic processes in scar tissue.
