Anti-inflammatory effects of anti-insulin resistant drugs and monocyte functional analysis in visceral obesity

本研究旨在探讨肥胖和代谢综合征患者脂肪细胞因子失调与单核细胞功能的关系以及抗胰岛素抵抗药物对单核细胞功能的影响，以阐明脂肪细胞-巨噬细胞相互作用的分子机制. THP-1细胞检测：THP-1细胞中，与PPAR配体、TZDs（吡格列酮6μM）共同孵育24 h可显著降低IL-6和MMP-9的表达，与EPA 50 μM共同孵育24 h可显著降低CD 14、TNFα、MCP-1、MMP-9和TLR 4的表达，并升高脂联素.使用人单核细胞的检查：我们在通过MACS或Ficoll-Paque方法从对照、肥胖和糖尿病的血液中分离外周血单核细胞后检查单核细胞功能。(1)实时荧光定量PCR检测单核细胞TNF-α、IL-6和MCP-1的表达，肥胖组和糖尿病组均高于对照组。(2)流式细胞仪检测到的单核细胞特异性标志物CD 14的表达和活性在对照组和肥胖组之间无显著差异，而在糖尿病患者中，CD 14和失活的M2巨噬细胞的表达和活性显著降低。(3)TZDs治疗3个月后，TNF-α的表达和CD 14的活性明显升高。临床研究：我们揭示了高度纯化的EPA减少小而密的LDL（sdLDL）、残余物和CRP（Diabetes Care 27：144，2007），并增加肥胖啮齿动物模型和人类肥胖受试者中的脂联素分泌，这可能是通过改善肥胖相关的脂肪组织炎症（ATVB 27：1918，2007）。

In the present study, to elucidate the molecular mechanisms of adipocyte-macrohphage interaction, we examined the relationship between adipocytokine dysregulation and the function of monocyte and the effect of anti-insulin resistant agents on the function of monocyte in the obesity and metabolic syndrome.1. Examination using the THP-1 cell : In THP-1 cell, the incubation with of PPAR ligand ; TZDs(Pioglitazone 6μM) for 24 hours significantly decreased the expression of IL-6 and MMP-9 and the administration of EPA 50 μM for 24 hours significantly decreased CD14, TNFα, MCP-1, MMP-9 and TLR4 and increased adiponectin.2. Examination using the human monocyte : We examined monocyte function after separating peripheral blood monocyte cells by MACS or Ficoll-Paque method from blood in control, obesity and diabetes. (1) The expressions of TNF-α, IL-6 and MCP-1 detected by real-time PCR in monocyte were higher in obesity and diabetes compared to control. (2) there is no significant difference between the expression and activity of CD14, a specific marker of monocyte, detected by FACS between control and obesity, however in diabetic patients the expression and activity of CD14 and inactive M2 Macrophage was significantly lower. (3) Administrating TZDs for 3 months, the expression of TNF-α and CD14 activity were improved3. Clinical Study : We revealed that highly purified EPA reduces small, dense LDL(sdLDL), remnants, and CRP(Diabetes Care 27 : 144, 2007) and increases adiponectin secretion in rodent models of obesity and human obese subjects, possibly through the improvement of obese-related adipose tissue inflammation(ATVB 27 : 1918, 2007).