Laboratory of Cell-Death Control BioTechnology, Faculty of Life and Environmental Sciences, Prefectural University of Hiroshima, Shobara, Hiroshima, Japan.
Free Radic Biol Med. 2011 Oct 1;51(7):1376-89. doi: 10.1016/j.freeradbiomed.2011.05.026. Epub 2011 May 27.
Adipose tissue is a crucial site for pathologic changes in obesity/metabolic syndrome-related diseases. Interaction between adipogenesis and reactive oxygen species (ROS) in adipose tissue involving chronic low-grade inflammation is postulated to be causal in the development of insulin resistance and other metabolic consequences. We used different culture systems to investigate the relationship between ROS and adipogenesis at three levels: within adipocytes, during adipocyte-monocyte interactions, and in a subcutaneous adipose tissue model. The effects of highly hydroxylated fullerene (HHF; C(60)(OH)(36)) on adipogenesis-accompanying oxidative stress and inflammatory changes were examined using these three systems. We demonstrated that H(2)O(2) stimulates lipid accumulation in 3T3-L1 preadipocytes, and lipid uptake causes ROS generation in OP9 preadipocytes, both of which were then markedly suppressed with HHF treatment. HHF significantly inhibited the adipogenic stimulant insulin-rich serum replacement (SR)-induced triacylglycerol accumulation, ROS production, and macrophage activation in cultured OP9 cells and an OP9-U937 monocyte-like cell coculture system. H(2)O(2)-induced intracellular ROS production in OP9 adipocytes was also notably inhibited by HHF. We developed a three-dimensional subcutaneous adipose-tissue equivalent (SATE) consisting of air-exposed cultures of HaCaT keratinocytes on an OP9 adipocyte-populated collagen gel in a culture insert. With SR stimulation and under suitable conditions, fat accumulation, ROS generation, and macrophage infiltration were observed in the SATE and significantly inhibited by HHF. By western blotting, we demonstrated that HHF localized at the cytoskeleton, which controls the transport of lipids. In conclusion, HHF is able to inhibit oxidative stress in adipocytes and adipogenesis-related macrophage activation in adipose tissues through its antioxidation.
脂肪组织是肥胖/代谢综合征相关疾病病理变化的关键部位。脂肪组织中脂肪生成和活性氧（ROS）之间的相互作用涉及慢性低度炎症，被认为是胰岛素抵抗和其他代谢后果发展的原因。我们使用不同的培养系统在三个层面上研究 ROS 与脂肪生成之间的关系：在脂肪细胞内、在脂肪细胞-单核细胞相互作用过程中、以及在皮下脂肪组织模型中。我们使用这三种系统研究了高度羟基化富勒烯（HHF；C(60)(OH)(36)）对伴随脂肪生成的氧化应激和炎症变化的影响。我们证明 H(2)O(2)刺激 3T3-L1 前脂肪细胞中的脂质积累，脂质摄取导致 OP9 前脂肪细胞中的 ROS 生成，而这两种情况都被 HHF 处理显著抑制。HHF 显著抑制了培养的 OP9 细胞和 OP9-U937 单核细胞样细胞共培养系统中胰岛素丰富的血清替代物（SR）诱导的三酰基甘油积累、ROS 产生和巨噬细胞激活。HHF 还显著抑制了 H(2)O(2)诱导的 OP9 脂肪细胞内的 ROS 生成。我们开发了一种三维皮下脂肪组织等效物（SATE），由暴露于空气中的 HaCaT 角质形成细胞在培养插入物中的 OP9 脂肪细胞填充的胶原凝胶上培养而成。在 SR 刺激下，并在适当的条件下，在 SATE 中观察到脂肪积累、ROS 生成和巨噬细胞浸润，并被 HHF 显著抑制。通过 Western 印迹，我们证明 HHF 定位于细胞骨架，它控制着脂质的运输。总之，HHF 能够通过其抗氧化作用抑制脂肪细胞中的氧化应激和脂肪生成相关的巨噬细胞激活。
