The Key Laboratory of Myocardial Ischemia Organization, Chinese Ministry of Education, Harbin, China; Department of Cardiology Organization, The Second Affiliated Hospital of Harbin Medical University, Harbin, China.
Department of Cardiology, Harbin Yinghua Hospital, Harbin, China.
Exp Cell Res. 2022 Dec 15;421(2):113407. doi: 10.1016/j.yexcr.2022.113407. Epub 2022 Nov 2.
Vascular calcification (VC) is closely related to higher cardiovascular mortality and morbidity, and vascular smooth muscle cell (VSMC) switching to osteogenic-like cells is crucial for VC. LncRNA LEF1-AS1 promotes atherosclerosis and dental pulp stem cells calcification, while its role in VC remains unknown. Visceral adipose tissue-derived serine protease inhibitor (vaspin) is an adipokine regulating bone metabolism. However, the relationship between vaspin and VC is still unclear. We aimed to explore the role of LEF1-AS1 on VSMC osteogenic transition, whether vaspin inhibited LEF1-AS1-mediated osteogenic differentiation of VSMCs, and the responsible mechanism. In this study, quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting analysis indicated that LEF1-AS1 overexpression significantly upregulated osteogenic marker Runt-related transcription factor-2 (RUNX2) level and downregulated VSMC contractile marker α-smooth muscle actin (α-SMA) level. Alizarin red staining, alkaline phosphatase (ALP) staining, ALP activity assay, and calcium content assay also suggested that LEF1-AS1 overexpression promoted calcium deposition in VSMCs. However, vaspin treatment abolished this phenomenon. Mechanistically, LEF1-AS1 markedly decreased phosphorylated YAP level, while vaspin reversed LEF1-AS1-induced phosphorylated YAP decline. Our results revealed that LEF1-AS1 accelerated the osteogenic differentiation of VSMCs by regulating the Hippo/YAP pathway, while vaspin eliminated the LEF1-AS1-meditated VSMCs osteogenic phenotype switch.
血管钙化（VC）与心血管死亡率和发病率升高密切相关，血管平滑肌细胞（VSMC）向成骨样细胞的转化对于 VC 至关重要。长链非编码 RNA LEF1-AS1 促进动脉粥样硬化和牙髓干细胞钙化，但其在 VC 中的作用尚不清楚。内脏脂肪组织来源的丝氨酸蛋白酶抑制剂（vaspin）是一种调节骨代谢的脂肪细胞因子。然而，vaspin 与 VC 之间的关系尚不清楚。本研究旨在探讨 LEF1-AS1 对 VSMC 成骨样转化的作用，vaspin 是否抑制了 LEF1-AS1 介导的 VSMCs 成骨分化，以及负责的机制。在这项研究中，定量实时聚合酶链反应（qRT-PCR）和 Western blot 分析表明，LEF1-AS1 的过表达显著上调成骨标志物 runt 相关转录因子 2（RUNX2）的水平，并下调 VSMC 收缩标志物α-平滑肌肌动蛋白（α-SMA）的水平。茜素红染色、碱性磷酸酶（ALP）染色、ALP 活性测定和钙含量测定也表明，LEF1-AS1 的过表达促进了 VSMCs 的钙沉积。然而，vaspin 的处理消除了这种现象。在机制上，LEF1-AS1 明显降低了磷酸化 YAP 的水平，而 vaspin 逆转了 LEF1-AS1 诱导的磷酸化 YAP 下降。我们的结果表明，LEF1-AS1 通过调节 Hippo/YAP 通路加速了 VSMCs 的成骨分化，而 vaspin 消除了 LEF1-AS1 介导的 VSMCs 成骨表型转换。
