Inducible toll-like receptor and NF-κB regulatory pathway expression in human adipose tissue
Inducible toll-like receptor and NF-κB regulatory pathway expression in human adipose tissue
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DOI：
10.1038/oby.2008.25
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发表时间：
2008-05-01
期刊：
影响因子：
6.9
通讯作者：
Gokce, Noyan
中科院分区：
文献类型：
--
作者：
Vitseva, Olga I.;Tanriverdi, Kahraman;Gokce, Noyan
目的：脂肪组织中的炎症活性最近被认为与胰岛素抵抗和肥胖相关的代谢功能障碍的机制有关。Toll样受体（TLR）在先天免疫应答中起关键作用，最近的研究表明TLR途径参与炎症和动脉粥样硬化的机制。本研究的目的是研究人类脂肪组织中TLR的差异表达和功能。方法和步骤：我们对16名肥胖受试者（年龄39 +/- 11岁，BMI 49 +/- 14 kg/m2）的皮下腹部脂肪进行活检，并使用定量实时PCR和共聚焦免疫荧光成像来表征TLR的表达。在组织培养中，我们用Pam 3CSK 4和脂多糖（LPS）刺激分离的人脂肪细胞，（分别为TLR 2和TLR 4激动剂）和定量TLR活性、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的产生和核因子-κ B（NF-κ B）p65核活化使用实时PCR、酶联免疫吸附测定（ELISA）和免疫荧光。TLR 1、2和4蛋白在人脂肪细胞中与脂联素共定位，其中TLR 4表现出最高的免疫组化表达。使用实时PCR，我们证实了与TLR家族的其他成员（TLR 1、2、7、8）相比，TLR 4在人脂肪库中的基因表达水平更高（P < 0.001）。在组织培养中，Pam 3CSK 4和LPS作用后，脂肪细胞TLR 2/TLR 4 mRNA和蛋白表达均显著增加（P < 0.001）。TLR 2/TLR 4刺激与NF-κ B B p65核转位和促炎性细胞因子production.Discussion：研究结果表明，TLR是诱导脂肪组织和下游NF-κ B激活和细胞因子释放。脂肪储存可能通过调节TLR/NF-κ B调节途径在人类受试者的炎症和先天免疫调节中发挥动态作用。
Objective: Inflammatory activity in fat tissue has recently been implicated in mechanisms of insulin resistance and obesity-related metabolic dysfunction. Toll-like receptors (TLRs) play a key role in innate immune responses and recent studies implicate the TLR pathway in mechanisms of inflammation and atherosclerosis. The aim of this study was to examine differential TLR expression and function in human adipose tissue.Methods and Procedures: We biopsied subcutaneous abdominal fat from 16 obese subjects (age 39 +/- 11 years, BMI 49 +/- 14 kg/m(2)) and characterized TLR expression using quantitative real-time PCR and confocal immunofluorescence imaging. In tissue culture, we stimulated isolated human adipocytes with Pam3CSK4 and lipopolysaccharide (LPS) (TLR2 and TLR4 agonists, respectively) and quantified TLR activity, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) production, and nuclear factor-kappa B (NF-kappa B) p65 nuclear activation using real-time PCR, enzyme-linked immunosorbent assay (ELISA), and immunofluorescence.Results: TLR1, 2, and 4 protein colocalized with adiponectin in human adipocytes with TLR4 exhibiting the highest immunohistochemical expression. Using real-time PCR, we confirmed higher level of gene expression for TLR4 as compared to other members of the TLR family (TLR1, 2, 7, 8) in human adipose depots (P < 0.001). In tissue culture, adipocyte TLR2/TLR4 mRNA expression and protein increased significantly following Pam3CSK4 and LPS (P < 0.001). TLR2/TLR4 stimulation was associated with NF-kappa B p65 nuclear translocation and proinflammatory cytokine production.Discussion: The findings demonstrate that TLRs are inducible in adipose tissue and linked with downstream NF-kappa B activation and cytokine release. Adipose stores may play a dynamic role in the regulation of inflammation and innate immunity in human subjects via modulation of the TLR/NF-kappa B regulatory pathway.