低剂量晚期糖基化终产物修饰低密度脂蛋白改善小鼠骨髓来源肥大 .docVIP

晚期糖基化终产物修饰的低密度脂蛋白下调小鼠骨髓来源肥大细胞IL-6和TNF-α释放 黄骁，赵馨娜，梁春，吴宗贵 摘要 目的 观察AGE-LDL对LPS诱导小鼠骨髓来源肥大细胞(Bone marrow derived mast cells, mBMMCs)释放IL-6和TNF-α的影响，并探讨HO-1对mBMMCs IL-6和TNF-α释放的影响。方法 分别用LPS(100ng/ml)、LPS联合AGE-LDL（50ug/ml）、LPS联合n-LDL(50ug/ml)刺激mBMMCs，另设对照组，并用ELISA试剂盒检测上清中IL-6和TNF-α浓度，real-time PCR检测mBMMCs HO-1mRNA表达水平，western blot检测MAPK p-P38、p-ERK1/2和NF-κB p-P65蛋白表达水平。在各组加用HO-1抑制剂Znpp，并检测上清IL-6和TNF-α浓度。结果 AGE-LDL 可显著下调LPS诱导mBMMCs释放的IL-6和TNF-α（p0.001, p0.001）。Znpp使AGE-LDL对mBMMCs释放IL-6和TNF-α的下调作用消失。AGE-LDL可诱导mBMMCs MAPK p-P38、p-ERK1/2和NF-κB p-P65蛋白表达,不影响LPS诱导的MAPK p-P38、p-ERK1/2和NF-κB p-P65蛋白表达升高。结论 AGE-LDL可下调LPS诱导mBMMCs释放的IL-6和TNF-α，此下调是通过升高HO-1表达而起作用。AGE-LDL可激活MAPK P38及ERK和NF-κB信号通路。AGE-LDL并非通过抑制MAPK和NF-κB信号通路，下调LPS诱导mBMMCs释放IL-6和TNF-α。 关键词 晚期糖基化终产物；低密度脂蛋白；肥大细胞；白细胞介素6；肿瘤坏死因子α Advanced glycation end products modified low density lipoprotein (AGE-LDL) down-regulated LPS-induced IL-6 and TNF-α release by murine bone marrow derived mast cells(mBMMCs). Abstract Objective To investigate the effect of AGE-LDL on LPS-induced IL-6 and TNF-α release by mBMMCs. Methods mBMMCs were cultured with n-LDL(50ug/ml), AGE-LDL(50ug/ml), LPS(100ng/ml), LPS(100ng/ml) plus n-LDL(50ug/ml) and LPS(100ng/ml) plus AGE-LDL(50ug/ml), respectively. Level of IL-6 and TNF-αin supernatant were measured by ELISA. HO-1 mRNA expression was measured by real time PCR. MAPK p-P38、p-ERK1/2 and NF-κB p-P65 protein expression were measured by western blot. Znpp, an HO-1 inhibitor, was added to each group, and level of IL-6 and TNF-αin supernatant were measured by ELISA. Results AGE-LDL significantly down-regulated LPS-induced IL-6 and TNF-α release by mBMMCs（p0.001, p0.001）. The down-regulated effect of AGE-LDL on LPS-induced IL-6 and TNF-α release by mBMMCs was diminished by Znpp. AGE-LDL promoted MAPK p-P38、p-ERK1/2 and NF-κB p-P65 protein expression in mBMMCs. However, AGE-LDL did not affect LPS-induced MAPK p-P38、p-ERK1/2 and NF-κB p-P65 protein expression in mBMMCs. Conclusion AGE-LDL down-regulated LPS-ind