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Inhibition of the PI 3 K / AKT Pathway Reduces Tumor Necrosis Factor‐Alpha Production in the Cellular Response to Wear Particles In Vitro
Author(s) -
Huang Jianbin,
Ding Yue,
Huang Dongsheng,
Liang Anjing,
Zeng Weike,
Zeng Zhanpeng,
Qin Chuqiang,
Barden Bertram
Publication year - 2013
Publication title -
artificial organs
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.684
H-Index - 76
eISSN - 1525-1594
pISSN - 0160-564X
DOI - 10.1111/j.1525-1594.2012.01568.x
Subject(s) - tumor necrosis factor alpha , protein kinase b , proinflammatory cytokine , secretion , stimulation , inflammation , chemistry , macrophage , microbiology and biotechnology , cancer research , signal transduction , biology , immunology , in vitro , biochemistry , endocrinology
Joint replacement is the most effective treatment for end‐stage osteoarticular disease. However, macrophage‐mediated aseptic loosening of joint prosthesis severely hampers the clinical effects of joint replacement. Until now, the mechanism by which macrophages regulate the secretion of inflammatory cytokines after particle stimulation is not clear. It is well known that the PI 3 K / AKT pathway participates in multiple cellular processes, including cell growth, survival, and inflammation. However, whether the PI 3 K / AKT pathway participates in the proinflammatory response of macrophages after particle stimulation and secondary aseptic loosening is still unknown. In this study, ceramic and titanium particles of different sizes were prepared to stimulate macrophages. LY 294002, a specific inhibitor of PI 3 K , was pretreated prior to particle stimulation. The expression of tumor necrosis factor‐alpha ( TNF ‐α) and all the subunits of PI 3 K and AKT were detected by real‐time polymerase chain reaction, enzyme‐linked immunosorbent assay, and W estern blot. The result showed that LY 294002 could suppress the RNA and protein expression of TNF ‐α in RAW 264.7 cells after stimulation of different particles. The subunits of PI 3 K (p110β and p85β), followed by activation of phosphor‐ AKT ( S er473), participated in the regulation of activating macrophages by wear particles, ultimately resulting in the secretion of TNF ‐α.