SPI1 Regulates the Progression of Ankylosing Spondylitis by Modulating TLR5 via NF-κB Signaling

Ankylosing spondylitis (AS) is an autoimmune disease which associated with inflammation of the spinal joints. Enhanced osteogenic differentiation was observed in AS; however, the underlying mechanism remains undefined. A cohort of AS (n = 15) and patients with traumatic fracture (n = 15) were recruited to this study. Fibroblasts were isolated, and characterized by H&E and immunocytochemistry (ICC) analysis. The expression and secretion of key molecules were detected by qRT-PCR, western blot, immunofluorescence (IF), and ELISA. Calcium deposition and alkaline phosphatase (ALP) activity were monitored by Alizarin Red S and ALP staining. The direct association between Spi-1 proto-oncogene (SPI1) and toll-like receptor 5 (TLR5) promoter was assessed by ChIP assay. AS fibroblasts was successfully isolated and exhibited osteogenic differentiation potentials. SPI1 was elevated in AS fibroblasts, and silencing of SPI1 inhibited osteogenic differentiation of AS fibroblasts. Mechanistic study showed that SPI1 acted as a transcriptional activator of TLR5. Knockdown of TLR5 suppressed osteogenic differentiation of AS fibroblasts via nuclear factor kappa B (NF-?B) signaling. Rescue experiments revealed that overexpression of TLR5 reversed SPI1 knockdown-suppressed osteogenic differentiation via NF-?B signaling. SPI1 regulated the progression of AS by modulating TLR5 via NF-?B signaling.

Automated summary

AS fibroblasts exhibited enhanced osteogenic differentiation potentials, with elevated expression of SPI1. Silencing of SPI1 inhibited osteogenic differentiation, while overexpression of TLR5 reversed this inhibition via NF-κB signaling. SPI1 regulated AS progression by modulating TLR5 through NF-κB signaling.