Kidney Disease Center, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Institute of Nephrology, Zhejiang University, Hangzhou, China.
Pharm Biol. 2022 Dec;60(1):17-24. doi: 10.1080/13880209.2021.2007268.
Vascular calcification is a major complication of chronic renal failure, which has been identified as an active process partly driven by osteogenic transition of vascular smooth muscle cells (VSMCs). Aspirin could prevent cardiomyocyte damage by inducing heat shock response.
This study investigates the effect of aspirin on alleviating VSMC calcification.
An VSMC calcification model was established by 10-day calcification induction in osteogenic medium. VSMCs were grouped as following: control group (normal medium), calcified group (osteogenic medium) and treated group (osteogenic medium with 1 or 4 mmol/L aspirin). VSMC calcification was evaluated by calcified nodules formation, intracellular calcium concentration and osteoblastic marker (OPN and Runx2) expression.
After 10-day culture, the intracellular calcium concentration in calcified group was significantly higher than that in control group (1.16 ± 0.04 vs. 0.14 ± 0.01 μg/mg, < 0.01), but significantly reduced in 1 mmol/L aspirin treated group (0.74 ± 0.05 μg/mg, < 0.01), and 4 mmol/L aspirin treated group (0.93 ± 0.03 μg/mg, < 0.01). The elevated expression of OPN and Runx2 induced by osteogenic medium was significantly relieved after 1 or 4 mmol/L aspirin treatment. The expression of HSF1, HSP70 and HSP90 was decreased in calcification-induced VSMCs, but significantly increased after treatment of aspirin. Furthermore, inhibition of HSP70 (or HSP90) by small-molecule inhibitor or small interfering RNA could partially abolish the anti-calcification effect of aspirin, proved by the changes of intracellular calcium concentration and osteoblastic marker expression.
Aspirin could relieve the calcification of VSMCs partially through HSP70- or HSP90-mediated heat shock response. These findings expanded the understanding of aspirin pharmacology, and imply that local induction expression of HSPs might be a potential therapeutic strategy for the prevention and therapy of vascular calcification.
血管钙化是慢性肾衰竭的主要并发症，该过程被认为是血管平滑肌细胞（VSMCs）发生成骨样转变的主动过程。阿司匹林可通过诱导热休克反应预防心肌细胞损伤。
本研究旨在探讨阿司匹林对减轻 VSMC 钙化的作用。
采用成骨诱导培养基培养 10 天建立 VSMC 钙化模型。将 VSMCs 分为以下几组：对照组（正常培养基）、钙化组（成骨培养基）和治疗组（成骨培养基加 1 或 4mmol/L 阿司匹林）。通过钙化结节形成、细胞内钙浓度和成骨细胞标志物（OPN 和 Runx2）表达评估 VSMC 钙化。
培养 10 天后，钙化组细胞内钙浓度明显高于对照组（1.16±0.04 vs. 0.14±0.01μg/mg， < 0.01），但在 1mmol/L 阿司匹林治疗组（0.74±0.05μg/mg， < 0.01）和 4mmol/L 阿司匹林治疗组（0.93±0.03μg/mg， < 0.01）明显降低。成骨培养基诱导的 OPN 和 Runx2 表达升高，经 1 或 4mmol/L 阿司匹林治疗后明显缓解。热休克蛋白（HSP）70 和 HSP90 的表达在钙化诱导的 VSMCs 中降低，但经阿司匹林处理后明显增加。此外，HSP70（或 HSP90）的小分子抑制剂或小干扰 RNA 抑制可部分消除阿司匹林的抗钙化作用，这可通过细胞内钙浓度和成骨细胞标志物表达的变化得到证实。
阿司匹林可部分通过 HSP70 或 HSP90 介导的热休克反应缓解 VSMCs 的钙化。这些发现扩展了对阿司匹林药理学的认识，提示局部诱导 HSPs 的表达可能是预防和治疗血管钙化的潜在治疗策略。
