Micromechanical force promotes aortic valvular calcification
Micromechanical force promotes aortic valvular calcification
复制标题
微机械力促进主动脉瓣钙化
DOI：
10.1016/j.jtcvs.2021.08.014
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发表时间：
2022-11-11
影响因子：
6
通讯作者：
Dong, Nianguo
中科院分区：
文献类型：
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作者：
Jiang, Yefan;Chen, Jinjie;Dong, Nianguo
目的：钙化性主动脉瓣病变是一种与力相关的炎症过程。本研究旨在探讨微机械力是否能诱导猪瓣膜间质细胞的瓣膜钙化，并采用磁扭转细胞仪检测整合素α v β 3在瓣膜钙化中的作用。检测钙化相关因子骨桥蛋白和RUNX 2的变化。利用该钙化介质，确定了诱导瓣膜间质细胞钙化的最佳磁旋流式细胞术参数，建立了磁旋流式细胞术钙化促进模型。用α v β 3拮抗剂阻断α v β 3的功能，研究α v β 3在钙化过程中的作用。结果：25 G-1Hz刺激5 min后，骨桥蛋白的表达增强。Western blotting结果显示25 G-1Hz刺激5分钟后24小时骨桥蛋白和RUNX 2的表达上调。诱导猪瓣膜间质细胞钙化的最佳磁扭转流式细胞术参数为25 G-2 Hz，10分钟。加入α v β 3拮抗剂后，骨桥蛋白和RUNX 2的表达明显降低。临床上，二叶式主动脉瓣患者主动脉瓣中RUNX 2和β 3表达较高，且β 3与RUNX 2表达显著相关。结论：利用磁扭转流式细胞术建立了微机械力刺激下猪主动脉瓣间质细胞钙化模型，并获得了最佳参数。整合素avb 3在主动脉瓣钙化过程中起关键作用。
Objective: Calcified aortic valvular disease is known as an inflammation-related process related to force. The purpose of this study was to determine whether micromechanical force could induce valve calcification of porcine valvular interstitial cells and to examine the role of integrin alpha v beta 3 in valvular calcification by using a novel method: magnetic twisting cytometry.Methods: Porcine valvular interstitial cells were cultured in vitro, and micromechanical force was applied to porcine valvular interstitial cells using magnetic twisting cytometry. Changes in calcification-related factors osteopontin and RUNX2 were detected. By using the calcification medium, the optimal magnetic twisting cytometry parameters for inducing valvular interstitial cell calcification were determined, and a magnetic twisting cytometry calcification promotion model was established. The role of alpha v beta 3 in calcification was studied by using alpha v beta 3 antagonists to block the function of alpha v beta 3.Results: Reverse transcription polymerase chain reaction assays showed that the expression of osteopontin was enhanced 30 minutes after 25G-1Hz 5 minutes of stimulation. Western blotting assays showed that the expression of osteopontin and RUNX2 was upregulated 24 hours after 25G-1Hz 5 minutes of stimulation. The optimal magnetic twisting cytometry parameter for inducing porcine valvular interstitial cell calcification was 25G-2Hz for 10 minutes. The expression of osteopontin and RUNX2 decreased significantly after the addition of alpha v beta 3 antagonist. Clinically, patients with bicuspid aortic valves had high expression of RUNX2 and beta 3 in the aortic valve, and b3 significantly correlated with RUNX2.Conclusions: By using magnetic twisting cytometry, we established a porcine valvular interstitial cell calcification model by micromechanical force stimulation and obtained the optimal parameters. Integrin avb3 plays a key role in the aortic valve calcification process.