Department of Molecular Sciences, College of Medicine, University of Tennessee Health Science Center, Memphis, TN 38163, USA.
J Inflamm (Lond). 2010 Jan 21;7:4. doi: 10.1186/1476-9255-7-4.
The present studies were designed to investigate the changes in gene expression during in vitro incubation of human visceral omental adipose tissue explants as well as fat cells and nonfat cells derived from omental fat.
Adipose tissue was obtained from extremely obese women undergoing bariatric surgery. Explants of the tissue as well as fat cells and the nonfat cells derived by digestion with collagenase were incubated for 20 minutes to 48 h. The expression of interleukin 1beta [IL-1beta], tumor necrosis factor alpha [TNFalpha], interleukin 8 [IL-8], NFkappaB(1)p50 subunit, hypoxia-inducible factor 1alpha [HIF1alpha], omentin/intelectin, and 11beta-hydroxysteroid dehydrogenase 1 [11beta-HSD1] mRNA were measured by qPCR as well as the release of IL-8 and TNFalpha.
There was an inflammatory response at 2 h in explants of omental adipose tissue that was reduced but not abolished in the absence of albumin from the incubation buffer for IL-8, IL-1beta and TNFalpha. There was also an inflammatory response with regard to upregulation of HIF1alpha and NFkappaB1 gene expression that was unaffected whether albumin was present or absent from the medium. In the nonfat cells derived by a 2 h collagenase digestion of omental fat there was an inflammatory response comparable but not greater than that seen in tissue. The exception was HIF1alpha where the marked increase in gene expression was primarily seen in intact tissue. The inflammatory response was not seen with respect to omentin/intelectin. Over a subsequent 48 h incubation there was a marked increase in IL-8 mRNA expression and IL-8 release in adipose tissue explants that was also seen to the same extent in the nonfat cells incubated in the absence of fat cells.
The marked inflammatory response seen when human omental adipose tissue is incubated in vitro is reduced but not abolished in the presence of albumin with respect to IL-1beta, TNFalpha, IL-8, and is primarily in the nonfat cells of adipose tissue.
本研究旨在研究人内脏大网膜脂肪组织外植体以及从大网膜脂肪中分离得到的脂肪细胞和非脂肪细胞在体外孵育过程中基因表达的变化。
从接受减肥手术的极度肥胖女性中获取脂肪组织。组织外植体以及用胶原酶消化得到的脂肪细胞和非脂肪细胞孵育 20 分钟至 48 小时。通过 qPCR 测量白细胞介素 1β（IL-1β）、肿瘤坏死因子α（TNFα）、白细胞介素 8（IL-8）、NFκB（1）p50 亚基、缺氧诱导因子 1α（HIF1α）、网膜素/intelectin 和 11β-羟类固醇脱氢酶 1（11β-HSD1）mRNA 的表达以及 IL-8 和 TNFα 的释放。
在大网膜脂肪组织外植体中，孵育 2 小时后出现炎症反应，在孵育缓冲液中不添加白蛋白时，IL-8、IL-1β 和 TNFα 的炎症反应减少但未被消除。此外，HIF1α 和 NFκB1 基因表达的上调也存在炎症反应，无论培养基中是否存在白蛋白，这种反应都不受影响。在大网膜脂肪经 2 小时胶原酶消化得到的非脂肪细胞中，观察到与组织相似但程度较小的炎症反应。例外的是 HIF1α，其基因表达的显著增加主要发生在完整组织中。关于网膜素/intelectin 未观察到炎症反应。在随后的 48 小时孵育中，在脂肪组织外植体中观察到 IL-8 mRNA 表达和 IL-8 释放的显著增加，在没有脂肪细胞的情况下孵育的非脂肪细胞中也观察到同样程度的增加。
当人内脏大网膜脂肪组织在体外孵育时，观察到明显的炎症反应，在存在白蛋白的情况下，IL-1β、TNFα、IL-8 的炎症反应减少但未被消除，主要发生在脂肪组织的非脂肪细胞中。
