Expression of PADI4 in patients with ankylosing spondylitis and its role in mediating the effects of TNF-α on the proliferation and osteogenic differentiation of human mesenchymal stem cells.

Abstract

Peptidyl arginine deiminase, type IV (PADI4) plays an important role in inflammation and in the immune response, and it has been shown to be associated with rheumatoid arthritis, osteoarthritis and ankylosing spondylitis (AS). However, little is known about the precise role of PADI4 in the pathogenic process in vitro. In this study, we aimed to investigate the expression of PADI4 in the synovial tissue of patients with AS and to determine the potential effects of PADI4 on human mesenchymal stem cell (hMSC) proliferation and osteogenic differentiation under normal and pathological conditions. Synovial tissues were collected from 18 patients with AS and 11 control subjects. The results of reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis revealed that the expression of PADI4 was upregulated in the patients with AS. In the hMSCs, the protein expression of PADI4 was increased following treatment with tumor necrosis factor-α (TNF-α) in a dose- and time-dependent manner. MTT assay revealed that TNF-α promoted hMSC proliferation. In addition, we found that TNF-α promoted the osteogenic differentiation of hMSCs, as demonstrated by an increase in alkaline phosphatase (ALP) activity, as well as an increase in the expression of bone morphogenetic protein 2 (BMP-2), runt-related transcription factor 2 (Runx2) and Osterix. The hMSCs were transfected with PADI4 siRNA to silence PADI4 expression. We found that, under normal conditions, the silencing of PADI4 did not have any effect on hMSC proliferation or osteogenic differentiation. However, in the presence of TNF-α, hMSC proliferation and osteogenic differentiation were induced. These effects were attenuated by the silencing of PADI4. In conclusion, the findings of this study demonstrate that the expression of PADI4 differs between patients with AS and normal subjects. In addition, our data suggest that PADI4 plays a role in hMSC proliferation and differentiation, which are induced by TNF-α.