Department of Physiology, College of Medicine, Yeungnam University, Daegu, South Korea.
J Endocrinol. 2010 Oct;207(1):35-44. doi: 10.1677/JOE-10-0093. Epub 2010 Jul 30.
The present study examined whether adiponectin can inhibit palmitate-induced apoptosis, and also the associated mechanisms and signal transduction pathways in human umbilical vein endothelial cells. Cells treated with 500 μM palmitate for 48 h increased reactive oxygen species (ROS) generation and induced apoptosis. Treatment with antioxidant N-acetyl-L-cysteine (1 mM) and globular adiponectin (5 μg/ml) inhibited palmitate-induced ROS generation and apoptosis. The AMP-activated protein kinase (AMPK) activator 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR; 1 mM), and cAMP activators forskolin (10 μM) and cholera toxin (200 ng/ml) also displayed the same effects. The inhibitory effects of adiponectin on ROS generation and apoptosis were reversed by the AMPK inhibitor compound C (40 μM), cAMP inhibitor SQ22536 (50 μM), and protein kinase A (PKA) inhibitor H-89 (10 μM). The inhibitory effect of forskolin on palmitate-induced apoptosis was reversed by compound C, whereas the inhibitory effect of AICAR was not reversed by SQ22536 and H-89. AICAR and forskolin could not inhibit palmitate-induced apoptosis in cells treated with dominant-negative AMPK. Forskolin increased phosphorylated AMPK at both Thr-172 and Ser-485/491. These results suggest that adiponectin inhibits palmitate-induced apoptosis by suppression of ROS generation via both the cAMP/PKA and AMPK pathways. Interaction between cAMP/PKA and AMPK pathways may be involved.
本研究旨在探讨脂联素是否能抑制棕榈酸诱导的人脐静脉内皮细胞凋亡,并探讨相关的机制和信号转导通路。用 500μM 棕榈酸处理 48 小时的细胞会增加活性氧(ROS)的产生并诱导细胞凋亡。用抗氧化剂 N-乙酰-L-半胱氨酸(1mM)和球形脂联素(5μg/ml)处理能抑制棕榈酸诱导的 ROS 产生和细胞凋亡。AMP 激活蛋白激酶(AMPK)激活剂 5-氨基咪唑-4-甲酰胺-1-β-D-核糖呋喃糖苷(AICAR;1mM)和环磷酸腺苷(cAMP)激活剂 forskolin(10μM)及霍乱毒素(200ng/ml)也有同样的效果。脂联素对 ROS 产生和细胞凋亡的抑制作用可被 AMPK 抑制剂化合物 C(40μM)、cAMP 抑制剂 SQ22536(50μM)和蛋白激酶 A(PKA)抑制剂 H-89(10μM)逆转。化合物 C 能逆转 forskolin 对棕榈酸诱导的细胞凋亡的抑制作用,而 SQ22536 和 H-89 则不能逆转 AICAR 的抑制作用。AICAR 和 forskolin 不能抑制 AMPK 失活的细胞中由棕榈酸引起的细胞凋亡。 forskolin 增加了 Thr-172 和 Ser-485/491 磷酸化的 AMPK。这些结果表明,脂联素通过 cAMP/PKA 和 AMPK 途径抑制 ROS 生成来抑制棕榈酸诱导的细胞凋亡。cAMP/PKA 和 AMPK 途径之间可能存在相互作用。