Department of Medicine, Division of Diabetes and the Texas Diabetes Institute, University Health System, 7703 Floyd Curl Drive MS 7886, San Antonio, TX, 78229-3900, USA.
Department of Cell Systems and Anatomy, University of Texas Health San Antonio, San Antonio, TX, USA.
Mol Biol Rep. 2020 Sep;47(9):6561-6572. doi: 10.1007/s11033-020-05707-w. Epub 2020 Aug 12.
After confirmation of the presence of adiponectin (ADPN) receptors and intra-cellular binding proteins in coronary artery smooth muscle cells (VSMC), we tested the hypotheses that, in acute insulin resistance: (i) the activation/inactivation of metabolic and mitogenic insulin signaling pathways are inversely affected by ADPN and, (ii) changes in VSMC migration/proliferation rates correlate with signal activity/inactivity. In primary cultures of VSMC exposed to high glucose and palmitate plus insulin, the expression of PI-3 kinase (Akt and m-TOR), MAP-Kinase (Erk and p-38) molecules, and inflammatory markers (TLR-4 and IkB-α) were assessed with Western blot, in the absence/presence of AdipoRon (AR). Migration and proliferation rates were measured in similar experimental conditions. There were decreases of ~ 25% (p-Akt) and 40-60% (p-mTOR) expressions with high glucose/palmitate, which reversed when AR was added were. Elevations in p-Erk and p-p38 expressions were obliterated by AR. Although, no changes were detected with high glucose and palmitate, when AR was added, a decline in inflammatory activity was substantiated by a ~ 50% decrease in TLR-4 and 40-60% increase in IkBα expression. Functional assays showed 10-20% rise in VSMC proliferation with high glucose and palmitate, but addition of AR lead to 15-25% decline. The degree of VSMC migration was reduced with AR addition by ~ 15%, ~ 35% and 55%, in VSMC exposed to 5 mM, 25 mM glucose and 25 mM + 200 µM palmitate, respectively. Changes in intracellular molecular messaging in experiments mimicking acute insulin resistance suggest that anti-inflammatory and anti-atherogenic actions of ADPN in VSMC are mediated via insulin signaling pathways.
在确认冠状动脉平滑肌细胞 (VSMC) 中存在脂联素 (ADPN) 受体和细胞内结合蛋白后,我们检验了以下假设:在急性胰岛素抵抗时:(i) 代谢和有丝分裂胰岛素信号通路的激活/失活受到 ADPN 的反向影响,和 (ii) VSMC 迁移/增殖率的变化与信号活性/失活相关。在暴露于高葡萄糖和棕榈酸加胰岛素的 VSMC 原代培养物中,通过 Western blot 评估了 PI-3 激酶 (Akt 和 m-TOR)、MAP 激酶 (Erk 和 p-38) 分子和炎症标志物 (TLR-4 和 IkB-α) 的表达,在不存在/存在 AdipoRon (AR) 的情况下进行。在类似的实验条件下测量迁移和增殖率。高葡萄糖/棕榈酸导致 p-Akt 和 p-mTOR 表达减少约 25%(p-Akt)和 40-60%(p-mTOR),当添加 AR 时则逆转。AR 消除了 p-Erk 和 p-p38 表达的升高。虽然在高葡萄糖和棕榈酸存在的情况下没有检测到变化,但当添加 AR 时,TLR-4 表达下降约 50%,IkBα表达增加 40-60%,证实炎症活性下降。功能测定显示,高葡萄糖和棕榈酸使 VSMC 增殖增加 10-20%,但添加 AR 会导致增殖下降 15-25%。在暴露于 5 mM、25 mM 葡萄糖和 25 mM + 200 µM 棕榈酸的 VSMC 中,添加 AR 可使 VSMC 迁移程度分别减少约 15%、35%和 55%。在模拟急性胰岛素抵抗的实验中,细胞内分子信息传递的变化表明,ADPN 在 VSMC 中的抗炎和抗动脉粥样硬化作用是通过胰岛素信号通路介导的。