Department of Clinical Biochemistry, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva 84103, Israel.
Endocrinology. 2010 Sep;151(9):4247-56. doi: 10.1210/en.2010-0340. Epub 2010 Jul 21.
Central obesity is frequently associated with adipose tissue inflammation and hepatic insulin resistance. To identify potential individual mediators in this process, we used in vitro systems and assessed if insulin resistance in liver cells could be induced by secreted products from adipocytes preexposed to an inflammatory stimulus. Conditioned medium from 3T3-L1 adipocytes pretreated without (CM) or with TNFalpha (CM-TNFalpha) was used to treat Fao hepatoma cells. ELISAs were used to assess the concentration of several inflammatory mediators in CM-TNFalpha. CM-TNFalpha-treated Fao cells exhibited about 45% diminution in insulin-stimulated phosphorylation of insulin receptor, insulin receptor substrate proteins, protein kinase B, and glycogen synthase kinase-3 as compared with CM-treated cells, without changes in the total abundance of these protein. Insulin increased glycogenesis by 2-fold in CM-treated Fao cells but not in cells exposed to CM-TNFalpha. Expression of IL-1beta mRNA was elevated 3-fold in TNFalpha-treated adipocytes, and CM-TNFalpha had 10-fold higher concentrations of IL-1beta but not TNFalpha or IL-1alpha. IL-1beta directly induced insulin resistance in Fao, HepG2, and in primary rat hepatocytes. Moreover, when TNFalpha-induced secretion/production of IL-1beta from adipocytes was inhibited by the IL-1 converting enzyme (ICE-1) inhibitor II (Ac-YVAD-CMK), insulin resistance was prevented. Furthermore, liver-derived cells treated with IL-1 receptor antagonist were protected against insulin resistance induced by CM-TNFalpha. Finally, IL-1beta secretion from human omental fat explants correlated with body mass index (R(2) = 0.639, P < 0.01), and the resulting CM induced insulin resistance in HepG2 cells, inhibitable by IL-1 receptor antagonist. Our results suggest that adipocyte-derived IL-1beta may constitute a mediator in the perturbed cross talk between adipocytes and liver cells in response to adipose tissue inflammation.
中心性肥胖常伴有脂肪组织炎症和肝胰岛素抵抗。为了确定这一过程中的潜在个体介质,我们使用了体外系统,并评估了预先暴露于炎症刺激的脂肪细胞分泌产物是否可以诱导肝细胞胰岛素抵抗。用未处理(CM)或用 TNFalpha 预处理(CM-TNFalpha)的 3T3-L1 脂肪细胞的条件培养基处理 Fao 肝癌细胞。ELISA 用于评估 CM-TNFalpha 中几种炎症介质的浓度。与 CM 处理的细胞相比,CM-TNFalpha 处理的 Fao 细胞中胰岛素刺激的胰岛素受体、胰岛素受体底物蛋白、蛋白激酶 B 和糖原合酶激酶-3 的磷酸化减少了约 45%,而这些蛋白的总量没有变化。胰岛素使 CM 处理的 Fao 细胞的糖生成增加了 2 倍,但不能使暴露于 CM-TNFalpha 的细胞增加。TNFalpha 处理的脂肪细胞中 IL-1beta mRNA 的表达增加了 3 倍,CM-TNFalpha 中 IL-1beta 的浓度高 10 倍,但 TNFalpha 或 IL-1alpha 的浓度没有增加。IL-1beta 直接诱导 Fao、HepG2 和原代大鼠肝细胞的胰岛素抵抗。此外,当脂肪细胞中 TNFalpha 诱导的 IL-1beta 分泌/产生被 IL-1 转化酶(ICE-1)抑制剂 II(Ac-YVAD-CMK)抑制时,胰岛素抵抗得到预防。此外,用 IL-1 受体拮抗剂处理的肝源性细胞可防止 CM-TNFalpha 诱导的胰岛素抵抗。最后,人网膜脂肪外植体中 IL-1beta 的分泌与体重指数(R(2) = 0.639,P < 0.01)相关,由此产生的 CM 可诱导 HepG2 细胞的胰岛素抵抗,用 IL-1 受体拮抗剂可抑制这种作用。我们的结果表明,脂肪细胞衍生的 IL-1beta 可能构成脂肪组织炎症反应中脂肪细胞和肝细胞之间失调的细胞间通讯的介质。