Inhibition of the PI3K/AKT Pathway Reduces Tumor Necrosis Factor-Alpha Production in the Cellular Response to Wear Particles In Vitro
Inhibition of the PI3K/AKT Pathway Reduces Tumor Necrosis Factor-Alpha Production in the Cellular Response to Wear Particles In Vitro
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DOI：
10.1111/j.1525-1594.2012.01568.x
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发表时间：
2013-03-01
影响因子：
2.4
通讯作者：
Barden, Bertram
中科院分区：
文献类型：
--
作者：
Huang, Jian-bin;Ding, Yue;Barden, Bertram
关节置换术是治疗终末期骨关节疾病最有效的方法。然而，巨噬细胞介导的人工关节无菌性松动严重影响了关节置换术的临床效果。到目前为止，巨噬细胞在颗粒刺激后调节炎性细胞因子分泌的机制尚不清楚。众所周知，PI 3 K/AKT通路参与多种细胞过程，包括细胞生长、存活和炎症。然而，PI 3 K/AKT通路是否参与了颗粒刺激后巨噬细胞的促炎反应和继发性无菌性松动仍不清楚。在这项研究中，制备了不同尺寸的陶瓷和钛颗粒来刺激巨噬细胞。在粒子刺激之前预处理PI 3 K的特异性抑制剂LY 294002。采用实时荧光定量聚合酶链反应、酶联免疫吸附试验和Western blot检测肿瘤坏死因子-α（TNF-α）、PI 3 K和AKT各亚基的表达。结果表明，LY 294002能抑制不同颗粒物刺激后RAW264.7细胞TNF-α的RNA和蛋白表达。PI 3 K的亚基（p110和p85），随后是磷酸化AKT（Ser 473）的激活，参与了磨损颗粒激活巨噬细胞的调节，最终导致TNF-α的分泌。
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 PI3K/AKT pathway participates in multiple cellular processes, including cell growth, survival, and inflammation. However, whether the PI3K/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. LY294002, a specific inhibitor of PI3K, was pretreated prior to particle stimulation. The expression of tumor necrosis factor-alpha (TNF-) and all the subunits of PI3K and AKT were detected by real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blot. The result showed that LY294002 could suppress the RNA and protein expression of TNF- in RAW264.7 cells after stimulation of different particles. The subunits of PI3K (p110 and p85), followed by activation of phosphor-AKT (Ser473), participated in the regulation of activating macrophages by wear particles, ultimately resulting in the secretion of TNF-alpha.