米诺环素调控P2X7受体抑制BV-2细胞活化的研究

摘要

AIM: To explore the role of P2X7 receptor in inhibition of lipopolysaccharide (LPS)-stimulated BV-2 cell activation by minocycline .METHODS:BV-2 cells were divided into 5 groups:control group, LPS group, LPS+0.1 μmol/L Mino group, LPS+1 μmol/L Mino group and LPS+10 μmol/L Mino group.The expression of P2X7 re-ceptor was determined by real-time PCR and Western blotting .The levels of TNF-αand IL-1βin the microglia culture su-pernatants were measured by ELISA .The morphological changes of the cells were also observed .RESULTS: After ex-posed to LPS, the expression of P2X7 receptor increased in BV-2 cells at mRNA and protein levels .The concentrations of TNF-αand IL-1βin the microglia culture supernatants also increased .Meanwhile, 0.1~10μmol/L minocycline inhibited those changes in a dose-dependent manner .CONCLUSION:Minocycline inhibits the activation of microglia .The mecha-nism may be related to the P2X7 receptor.%目的：探索P2X7受体在米诺环素（Mino）抑制脂多糖（LPS）刺激的BV-2细胞活化中的作用。方法：将体外培养的BV-2细胞分为5组：空白对照组、LPS处理组、LPS＋0．1μmol／L Mino组、LPS＋1μmol／L Mino组和LPS＋10μmol／L Mino组。分组处理8 h后行real-time PCR检测P2X7受体mRNA表达；处理24 h后观察各组细胞形态学变化，Western blotting检测P2X7受体蛋白表达，取细胞培养液上清行ELISA检测TNF-α和IL-1β的分泌情况。结果：经LPS处理后，BV-2细胞P2X7受体mRNA及蛋白的表达均升高，细胞培养液上清的TNF-α和IL-1β表达升高，同时伴有形态学的改变，而0．1～10μmol／L Mino 能抑制这一趋势，差异有统计学意义（ P ＜0.01）。结论：Mino抑制BV-2细胞活化的机制可能与抑制P2X7受体的活性相关。