Nonenzymatic glycation of high-density lipoprotein impairs its anti-inflammatory effects in innate immunity
Nonenzymatic glycation of high-density lipoprotein impairs its anti-inflammatory effects in innate immunity
复制标题
高密度脂蛋白的非酶糖化损害其在先天免疫中的抗炎作用
DOI：
10.1002/dmrr.1297
复制
发表时间：
2012-02-01
影响因子：
8
通讯作者：
Zheng, Lemin
中科院分区：
文献类型：
--
作者：
Liu, Donghui;Ji, Liang;Zheng, Lemin
目的/假设2型糖尿病（T2 DM）患者高密度脂蛋白（HDL）的蛋白质和脂质组成异常，可损害其抗炎功能。方法将人急性单核细胞白血病细胞系（THP-1）、小鼠RAW 264.7巨噬细胞和原代人单核细胞衍生的巨噬细胞与天然HDL预孵育，从T2 DM患者分离的糖尿病HDL或HDL在体外用不同剂量的D-葡萄糖糖化，然后用脂多糖（LPS）攻击。通过酶联免疫吸附试验（ELISA）测定肿瘤坏死因子（TNF）-α和IL-1 β的释放。Western blot检测I κ-B α在细胞质中的磷酸化和NF-κ B的核转位。结果与天然HDL相比，糖尿病HDL抑制TNF-α（p < 0.05）和IL-1 β（p < 0.001）释放的能力明显减弱。类似地，体外HDL的糖化以葡萄糖剂量依赖性方式损害其抑制TNF-α和IL-1 β释放的能力。此外，apoHDL仍能有效抑制LPS诱导的TNF-α和IL-1 β的释放，但糖化apoHDL部分丧失了这种能力。糖化HDL和糖尿病HDL的非酶糖化水平增加28倍（p < 0.001）和4倍结论糖尿病HDL和体外糖化HDL均部分丧失了抑制LPS诱导的巨噬细胞释放细胞因子的保护作用，HDL蛋白质成分的非酶糖化在这些损伤中起关键作用。版权所有（C）2011约翰威利父子有限公司
Aims/hypothesis In type 2 diabetes mellitus (T2DM), the abnormal protein and lipid composition of diabetic high-density lipoprotein (HDL) could impair its anti-inflammatory functions. Whether nonenzymatic glycation directly impaired the anti-inflammatory effects of HDL in innate immunity remained unclear.Methods Human acute monocytic leukemia cell line (THP-1) cells, mouse RAW 264.7 macrophages and primary human monocytes derived macrophages were pre-incubated with native HDL, diabetic HDL isolated from T2DM patients or HDL glycated with different doses of D-glucose in vitro and then challenged with lipopolysaccharide (LPS). The release of tumor necrosis factor (TNF)-alpha and IL-1 beta was assayed by enzyme-linked immunosorbent assay (ELISA). Phosphorylation of I kappa-B alpha in cytoplasm and nuclear translocation of NF-kappa B were detected by western blot. Glycation levels of native HDL, glycated HDL and diabetic HDL were determined using LC-MS/MS.Results The potency of diabetic HDL to inhibit the release of TNF-alpha (p < 0.05) and IL-1 beta (p < 0.001) was dramatically attenuated compared with that of native HDL. Similarly, glycation of HDL in vitro impaired its ability to inhibit TNF-alpha and IL-1 beta release in a glucose dose-dependent manner. Moreover, apoHDL still effectively inhibited the release of TNF-alpha and IL-1 beta induced by LPS, but glycated apoHDL partly lost such abilities. Nonenzymatic glycation levels of glycated HDL and diabetic HDL increased 28 fold (p < 0.001) and 4 fold (p < 0.001), respectively compared with that of native HDL.Conclusions In this study, we observed that diabetic HDL and HDL glycated in vitro both partly lose their protective effects to inhibit cytokines release induced by LPS in macrophages, and nonenzymatic glycation of the protein components of HDL plays key roles in these impairments. Copyright (C) 2011 John Wiley & Sons, Ltd.