Bzatp induced expression and production of interleukin1 beta and tumor necrosis factor alpha by m2 macrophages from ankylosing spondylitis patients

Maryam Akhtari,1,* Seyed jalal zargar,2 Mahdi mahmoudi,3 Mahdi vojdanian,4 Ahmadreza jamshidi,5

1. School of Biology, College of Science, University of Tehran, Tehran, Iran
2. School of Biology, College of Science, University of Tehran, Tehran, Iran
3. Rheumatology Research Center, Tehran University of Medical Sciences, Tehran, Iran
4. Rheumatology Research Center, Tehran University of Medical Sciences, Tehran, Iran
5. Rheumatology Research Center, Tehran University of Medical Sciences, Tehran, Iran

Abstract

Introduction

Ankylosing spondylitis (as) is an auto-inflammatory spinal rheumatic disease. the disease that affects spinal column and sacroiliac joints is characterized by inflammatory type of low back pain and progressive spinal stiffness. inflammatory tissues contain populations of macrophages that involved in the auto-inflammatory responses and fibrocartilage destruction in as. extracellular adenosine triphosphate (atp), regulates several immune and inflammatory responses in macrophages by interaction with p2x7 purinergic receptor. the p2x7r is a receptor/channel that activation by extracellular atp causes the formation of inflammasome protein complex and release of pro-inflammatory cytokines. the effects of p2x7r activity on the pathogenesis of as is still unknown. in this study, we investigated (by bzatp) on the production of pro-inflammatory cytokines in m2 macrophages from healthy individuals and as patients.

Methods

Monocytes were collected from 14 healthy controls and 14 active as patients, differentiated to m2 macrophages by macrophage-colony stimulating factor (m-csf) for 7 days. m2 macrophages were then treated with bzatp (200 µm) for 24 h. analysis of tumor necrosis factor-alpha (tnf-a) and interleukin 1β (il-1β) mrna expression and protein production was performed using sybr green qpcr and elisa method.

Results

Mrna expression level of il-1β and tnf-α was significantly increased (p < 0.05) following treatment with bzatp in m2 macrophages from as patients and healthy controls. protein production of tnf-α was also significantly up-regulated (p < 0.01) following treatment with bzatp in m2 macrophages from as patients and normal individuals. m2 macrophages from as patients have a higher rate of tnf-α release (p < 0.05) by bzatp stimulation compared to healthy macrophages.

Conclusion

Our results suggested that bzatp can modulate inflammatory signaling more specifically in macrophages from as patients and has a less effect on healthy macrophages. therefor modulating the p2x7 receptor activation could be used as a therapeutic approach for as treatment.

Keywords

P2x7 receptor, m2 macrophage, ankylosing spondylitis