Research Centre for Healthy and Sustainable Living, Innovative Testing in Life Sciences and Chemistry, University of Applied Sciences, Heidelberglaan 7, 3584, CS, Utrecht, The Netherlands.
Laboratory of Biochemistry, Wageningen University, Dreijenlaan 3, 6703, HA, Wageningen, The Netherlands.
Arch Biochem Biophys. 2018 Apr 15;644:81-92. doi: 10.1016/j.abb.2018.03.003. Epub 2018 Mar 8.
Currently, there is a worldwide increase of patients with type 2 diabetes (T2D). During the progression of healthy obese to T2D status, there is an influx of immune cells, in particular macrophages, into visceral adipose tissue, accompanied by an increase of inflammatory cytokines, such as, IL6, TNFα and Hp. To get a better insight in the underlying mechanisms, we performed a quantitative LCMS analysis on a modified in vitro assay, combining 3T3L1 adipocytes and activated RAW264.7 macrophages, thus mimicking inflamed adipose tissue. Clinically known proteins, e.g. IL6, TNFα, AdipoQ, complement factor C3, B and D were identified, thus confirming the assay. In addition, we found 54 new proteins that can potentially be used for research into the mechanism of T2D. Comparison of our results to a study on human visceral fat of obese non-diabetic and obese diabetic subjects, indicated that AUH, NAGK, pCYT2, NNMT, STK39 and CSNK2A2 might indeed be linked to insulin resistance in humans. Moreover, the expression of some of these genes was also altered in human blood samples at early or later stages of insulin desensitization. Overall, we conclude that the direct contact co-culture of 3T3L1 adipocytes with activated macrophages could be a mechanistically relevant and partially translational model of inflamed visceral adipose tissue.
目前，全球 2 型糖尿病（T2D）患者人数不断增加。在健康肥胖人群向 T2D 状态发展的过程中，大量免疫细胞，尤其是巨噬细胞，涌入内脏脂肪组织，同时炎症细胞因子如 IL6、TNFα 和 Hp 的水平也会升高。为了更深入地了解潜在机制，我们在体外模拟炎症脂肪组织的改良共培养实验中对 3T3L1 脂肪细胞和激活的 RAW264.7 巨噬细胞进行了定量 LCMS 分析。该实验中鉴定到了临床上已知的蛋白，如 IL6、TNFα、AdipoQ、补体因子 C3、B 和 D，从而证实了该实验的可行性。此外，我们还发现了 54 种新的蛋白，它们可能被用于研究 T2D 的发病机制。将我们的结果与肥胖非糖尿病和肥胖糖尿病患者内脏脂肪的研究进行比较，表明 AUH、NAGK、pCYT2、NNMT、STK39 和 CSNK2A2 可能确实与人类的胰岛素抵抗有关。此外，这些基因的一些表达在人类血液样本中也在胰岛素脱敏的早期或晚期发生了改变。总的来说，我们得出结论，3T3L1 脂肪细胞与激活的巨噬细胞直接接触共培养可能是一种具有机制相关性且部分具有转化意义的炎症性内脏脂肪组织模型。
