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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
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).
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DOI:
作者:
黄骁,赵馨娜,梁春,吴宗贵
HUANG xiao, ZHAO xinna, LIANG chun, WU zonggui
作者单位:
上海长征医院
Shanghai Changzheng hospital
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
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(p<0.001, p<0.001)。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。
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(p<0.001, p<0.001). 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-induced IL-6 and TNF-α release by mast cells. The down-regulation was HO-1 dependent, but not MAPK and NF-κB pathways.
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