Department of Emergency and Chest Pain Center, Qilu Hospital of Shandong University, Jinan, China (X.Z., S.C., J.W., B.Q., X.L., R.H., M.Z., S.S., Y.H., S.W., J.P., Q.Y., F.X., S.W., Y.C.).
Shandong Provincial Clinical Research Center for Emergency and Critical Care Medicine, Institute of Emergency and Critical Care Medicine of Shandong University, Qilu Hospital of Shandong University, Jinan, China (X.Z., S.C., J.W., B.Q., X.L., R.H., M.Z., S.S., Y.H., S.W., J.P., Q.Y., F.X., S.W., Y.C.).
Circulation. 2024 May 28;149(22):1752-1769. doi: 10.1161/CIRCULATIONAHA.123.065830. Epub 2024 Feb 13.
Vascular calcification, which is characterized by calcium deposition in arterial walls and the osteochondrogenic differentiation of vascular smooth muscle cells, is an actively regulated process that involves complex mechanisms. Vascular calcification is associated with increased cardiovascular adverse events. The role of 4-hydroxynonenal (4-HNE), which is the most abundant stable product of lipid peroxidation, in vascular calcification has been poorly investigated.
Serum was collected from patients with chronic kidney disease and controls, and the levels of 4-HNE and 8-iso-prostaglandin F2α were measured. Sections of coronary atherosclerotic plaques from donors were immunostained to analyze calcium deposition and 4-HNE. A total of 658 patients with coronary artery disease who received coronary computed tomography angiography were recruited to analyze the relationship between coronary calcification and the rs671 mutation in aldehyde dehydrogenase 2 (). knockout () mice, smooth muscle cell-specific knockout mice, transgenic mice, and their controls were used to establish vascular calcification models. Primary mouse aortic smooth muscle cells and human aortic smooth muscle cells were exposed to medium containing β-glycerophosphate and CaCl to investigate cell calcification and the underlying molecular mechanisms.
Elevated 4-HNE levels were observed in the serum of patients with chronic kidney disease and model mice and were detected in calcified artery sections by immunostaining. knockout or smooth muscle cell-specific knockout accelerated the development of vascular calcification in model mice, whereas overexpression or activation prevented mouse vascular calcification and the osteochondrogenic differentiation of vascular smooth muscle cells. In patients with coronary artery disease, patients with rs671 gene mutation developed more severe coronary calcification. 4-HNE promoted calcification of both mouse aortic smooth muscle cells and human aortic smooth muscle cells and their osteochondrogenic differentiation in vitro. 4-HNE increased the level of Runx2 (runt-related transcription factor-2), and the effect of 4-HNE on promoting vascular smooth muscle cell calcification was ablated when Runx2 was knocked down. Mutation of Runx2 at lysine 176 reduced its carbonylation and eliminated the 4-HNE-induced upregulation of Runx2.
Our results suggest that 4-HNE increases Runx2 stabilization by directly carbonylating its K176 site and promotes vascular calcification. ALDH2 might be a potential target for the treatment of vascular calcification.
血管钙化的特征是钙在动脉壁中沉积和血管平滑肌细胞向成骨软骨样分化，是一个涉及复杂机制的主动调节过程。血管钙化与心血管不良事件的增加有关。脂质过氧化的最丰富稳定产物 4-羟基壬烯醛（4-HNE）在血管钙化中的作用尚未得到充分研究。
收集慢性肾脏病患者和对照者的血清，测定 4-HNE 和 8-异前列腺素 F2α的水平。对供体的冠状动脉粥样硬化斑块进行免疫染色，分析钙沉积和 4-HNE。共招募了 658 例接受冠状动脉计算机断层血管造影的冠状动脉疾病患者，分析冠状动脉钙化与醛脱氢酶 2（ALDH2）中 rs671 突变（）的关系。利用 基因敲除（）小鼠、血管平滑肌细胞特异性 基因敲除（）小鼠、 转基因（）小鼠及其对照小鼠建立血管钙化模型。将原代小鼠主动脉平滑肌细胞和人主动脉平滑肌细胞暴露于含有β-甘油磷酸和 CaCl 的培养基中，以研究细胞钙化和潜在的分子机制。
在慢性肾脏病患者和模型小鼠的血清中观察到 4-HNE 水平升高，并通过免疫染色在钙化动脉切片中检测到。 基因敲除或血管平滑肌细胞特异性 基因敲除加速了模型小鼠的血管钙化发展，而 过表达或激活则防止了小鼠血管钙化和血管平滑肌细胞的成骨软骨样分化。在冠状动脉疾病患者中，rs671 基因突变的患者发生更严重的冠状动脉钙化。4-HNE 促进了体外小鼠主动脉平滑肌细胞和人主动脉平滑肌细胞的钙化及其成骨软骨样分化。4-HNE 增加了 runt 相关转录因子 2（Runx2）的水平，当敲低 Runx2 时，4-HNE 促进血管平滑肌细胞钙化的作用被消除。Runx2 赖氨酸 176 的突变使其羰基化减少，并消除了 4-HNE 诱导的 Runx2 上调。
我们的结果表明，4-HNE 通过直接羰基化其 K176 位点增加 Runx2 的稳定性，并促进血管钙化。ALDH2 可能是治疗血管钙化的潜在靶点。
