Endocrinology and Metabolism, Department of Medicine and Surgery, University of Parma, Parma, Italy.
Department of Medicine and Surgery, Unit of Biomedical, Biotechnological and Translational Sciences (S.Bi.Bi.T.), University of Parma, Parma, Italy.
Cardiovasc Diabetol. 2020 Apr 7;19(1):46. doi: 10.1186/s12933-020-01016-5.
The clear evidence of cardiovascular benefits in cardiovascular outcome trials of sodium-glucose cotransporter 2 inhibitors (SGLT2i) in type 2 diabetes might suggest an effect on atherosclerotic plaque vulnerability and/or thrombosis, in which myeloid angiogenic cells (MAC) and platelets (PLT) are implicated. We tested the effects of SGLT2i on inflammation and oxidant stress in a model of stearic acid (SA)-induced lipotoxicity in MAC and on PLT activation. The possible involvement of the Na/H exchanger (NHE) was also explored.
MAC and PLT were isolated from peripheral blood of healthy subjects and incubated with/without SGLT2i [empagliflozin (EMPA) and dapagliflozin (DAPA) 1-100 μM] to assess their effects on SA (100 μM)-induced readouts of inflammation, oxidant stress and apoptosis in MAC and on expression of PLT activation markers by flow-cytometry after ADP-stimulation. Potential NHE involvement was tested with amiloride (aspecific NHE inhibitor) or cariporide (NHE1 inhibitor). Differences among culture conditions were identified using one-way ANOVA or Friedman test.
NHE isoforms (1,5-9), but not SGLT2 expression, were expressed in MAC and PLT. EMPA and DAPA (100 μM) significantly reduced SA-induced inflammation (IL1β, TNFα, MCP1), oxidant stress (SOD2, TXN, HO1), but not apoptosis in MAC. EMPA and DAPA (both 1 μM) reduced PLT activation (CD62p and PAC1 expression). SGLT2i effects were mimicked by amiloride, and only partially by cariporide, in MAC, and by both inhibitors in PLT.
EMPA and DAPA ameliorated lipotoxic damage in stearate-treated MAC, and reduced ADP-stimulated PLT activation, potentially via NHE-inhibition, thereby pointing to plaque stabilization and/or thrombosis inhibition as potential mechanism(s) involved in SGLT2i-mediated cardiovascular protection.
在 2 型糖尿病的心血管结局试验中，钠-葡萄糖共转运蛋白 2 抑制剂（SGLT2i）对心血管有明显益处，这可能表明其对动脉粥样硬化斑块的脆弱性和/或血栓形成有影响，其中髓系血管生成细胞（MAC）和血小板（PLT）起作用。我们在硬脂酸（SA）诱导的 MAC 脂毒性模型中测试了 SGLT2i 对炎症和氧化应激的影响，以及对 PLT 激活的影响。还探讨了钠/氢交换器（NHE）的可能参与。
从健康受试者的外周血中分离出 MAC 和 PLT，并与/或不与 SGLT2i[依帕列净（EMPA）和达格列净（DAPA）1-100 μM]孵育，以评估它们对 SA（100 μM）诱导的 MAC 炎症、氧化应激和细胞凋亡的影响，以及通过流式细胞术检测 ADP 刺激后 PLT 激活标志物的表达。用阿米洛利（非特异性 NHE 抑制剂）或 cariporide（NHE1 抑制剂）测试潜在的 NHE 参与。使用单向方差分析或 Friedman 检验识别培养条件之间的差异。
MAC 和 PLT 中表达 NHE 同工型（1、5-9），但不表达 SGLT2。EMPA 和 DAPA（100 μM）显著降低了 SA 诱导的 MAC 炎症（IL1β、TNFα、MCP1）、氧化应激（SOD2、TXN、HO1），但不降低细胞凋亡。EMPA 和 DAPA（均为 1 μM）降低了 PLT 激活（CD62p 和 PAC1 表达）。在 MAC 中，SGLT2i 的作用可被阿米洛利模拟，仅部分可被 cariporide 模拟，而在 PLT 中，两种抑制剂均可模拟。
EMPA 和 DAPA 改善了硬脂酸盐处理的 MAC 的脂毒性损伤，并降低了 ADP 刺激的 PLT 激活，可能通过 NHE 抑制，从而指出斑块稳定和/或血栓形成抑制作为 SGLT2i 介导的心血管保护的潜在机制。
