Institute for Diabetes and Obesity (IDO), Helmholtz Diabetes Center (HDC), Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), 85764 Oberschleissheim, Germany.
German Center for Diabetes Research (DZD), 85764 Oberschleissheim, Germany.
Molecules. 2020 Feb 18;25(4):896. doi: 10.3390/molecules25040896.
(1) Background: Acute administration of the cannabinoid receptor 1 (CB1R) inverse agonist Rimonabant (SR141716A) to fed Wistar rats was shown to elicit a rapid and short-lasting elevation of serum free fatty acids. (2) Methods: The effect of Rimonabant on lipolysis in isolated primary rat adipocytes was studied to raise the possibility for direct mechanisms not involving the (hypothalamic) CB1R. (3) Results: Incubation of these cells with Rimonabant-stimulated lipolysis to up to 25% of the maximal isoproterenol effect, which was based on both CB1R-dependent and independent mechanisms. The CB1R-dependent one was already effective at Rimonabant concentrations of less than 1 µM and after short-term incubation, partially additive to β-adrenergic agonists and blocked by insulin and, in part, by adenosine deaminase, but not by propranolol. It was accompanied by protein kinase A (PKA)-mediated association of hormone-sensitive lipase (HSL) with lipid droplets (LD) and dissociation of perilipin-1 from LD. The CB1R-independent stimulation of lipolysis was observed only at Rimonabant concentrations above 1 µM and after long-term incubation and was not affected by insulin. It was recapitulated by a cell-free system reconstituted with rat adipocyte LD and HSL. Rimonabant-induced cell-free lipolysis was not affected by PKA-mediated phosphorylation of LD and HSL, but abrogated by phospholipase digestion or emulsification of the LD. Furthermore, LD isolated from adipocytes and then treated with Rimonabant (>1 µM) were more efficient substrates for exogenously added HSL compared to control LD. The CB1R-independent lipolysis was also demonstrated in primary adipocytes from fed rats which had been treated with a single dose of Rimonabant (30 mg/kg). (4) Conclusions: These data argue for interaction of Rimonabant (at high concentrations) with both the LD surface and the CB1R of primary rat adipocytes, each leading to increased access of HSL to LD in phosphorylation-independent and dependent fashion, respectively. Both mechanisms may lead to direct and acute stimulation of lipolysis at peripheral tissues upon Rimonabant administration and represent targets for future obesity therapy which do not encompass the hypothalamic CB1R.
(1) 背景:研究表明,给喂食的 Wistar 大鼠急性施用大麻素受体 1(CB1R)反向激动剂利莫那班(SR141716A)会迅速短暂地升高血清游离脂肪酸。(2) 方法:研究利莫那班对分离的原代大鼠脂肪细胞脂解的影响,以提出可能涉及直接机制而非(下丘脑)CB1R 的可能性。(3) 结果:将这些细胞与利莫那班孵育可刺激脂解作用增加至最大异丙肾上腺素作用的 25%,这是基于 CB1R 依赖和非依赖机制。CB1R 依赖机制在利莫那班浓度小于 1µM 时就已经有效,并且在短期孵育后,部分与β-肾上腺素能激动剂相加,被胰岛素和部分腺苷脱氨酶阻断,但不受普萘洛尔影响。它伴随着激素敏感脂肪酶(HSL)与脂质滴(LD)的蛋白激酶 A(PKA)介导的结合以及 perilipin-1 从 LD 的解离。仅在利莫那班浓度高于 1µM 并经过长期孵育后才观察到 CB1R 非依赖性的脂解刺激,且不受胰岛素影响。它可以通过用大鼠脂肪细胞 LD 和 HSL 重建的无细胞系统再现。利莫那班诱导的无细胞脂解不受 PKA 介导的 LD 和 HSL 磷酸化影响,但被磷脂酶消化或 LD 乳化阻断。此外,从脂肪细胞分离并随后用利莫那班(>1µM)处理的 LD 与对照 LD 相比,更有效地成为外源添加的 HSL 的底物。在给予单剂量利莫那班(30mg/kg)的喂食大鼠的原代脂肪细胞中也观察到了 CB1R 非依赖性的脂解。(4) 结论:这些数据表明利莫那班(高浓度时)与原代大鼠脂肪细胞的 LD 表面和 CB1R 相互作用,分别导致 HSL 以非依赖和依赖磷酸化的方式更有效地进入 LD,从而分别导致脂解作用的直接和急性刺激。两种机制都可能导致利莫那班给药后外周组织的直接和急性脂解作用,并为不包括下丘脑 CB1R 的未来肥胖症治疗提供靶点。