Phosphoinositide 3-kinase β, phosphoinositide 3-kinase δ, and phosphoinositide 3-kinase γ mediate the anti-inflammatory effects of magnesium sulfate

Background: We previously demonstrated that inhibiting phosphoinositide 3-kinase (PI3K) or activating L-type calcium channels blocked the anti-inflammatory effects of magnesium sulfate (MgSO4). However, the question as which class I PI3K isoform (PI3K alpha, PI3K beta, PI3K delta, or PI3K gamma) is involved in this regard remains unstudied. The question as whether MgSO4 and L-type calcium channels interact to influence PI3K activation also remains unstudied. We therefore designed this study to test two hypotheses: (1) inhibiting PI3K alpha, PI3K beta, PI3K delta, or PI3K gamma would block the anti-inflammatory effects of MgSO4 and (2) activating L-type calcium channels would block the effects of MgSO4 on activating PI3K. Materials and methods: PI3K isoform investigation: macrophages (RAW264.7 cells) were treated with endotoxin, endotoxin plus MgSO4, or endotoxin plus MgSO4 plus the selective inhibitor of PI3K alpha (PIK-75), PI3K beta (TGX-221), PI3K delta (IC-87114), or PI3K gamma (AS-252424). Calcium channel investigation: macrophages were treated with endotoxin, endotoxin plus MgSO4, or endotoxin plus MgSO4 plus the L-type calcium channel activator BAY-K8644. Results: The endotoxin plus MgSO4 group presented lower concentrations of inflammatory mediators (macrophage inflammatory protein 2, tumor necrosis factor a, and interleukin 6, lower nuclear concentration of phosphorylated nuclear factor kappa B, lower cytosolic concentration of phosphorylated inhibitor kappa B alpha, and higher concentration of phosphorylated Akt (PI3K activation marker) than the endotoxin group (all P < 0.05). These effects of MgSO4 were significantly reduced by TGX-221, IC-87114, or AS-252424, but not PIK-75. Additionally, BAY-K8644 blocked the effect of MgSO4 on activating PI3K. Conclusions: MgSO4 exerts its anti-inflammatory effects through activating PI3K beta, PI3K delta, and PI3K gamma. The underlying mechanism appears to involve inhibition of L-type calcium channels. (C) 2015 Elsevier Inc. All rights reserved.