The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Roslin Biocentre, Roslin, UK.
Int J Mol Med. 2011 May;27(5):663-8. doi: 10.3892/ijmm.2011.631. Epub 2011 Mar 1.
Vascular calcification has severe clinical consequences in a number of diseases, including diabetes, atherosclerosis and end-stage renal disease. The in vitro calcification of primary mouse, human and bovine vascular smooth muscle cells (VSMCs) is commonly employed to examine the mechanisms of vascular calcification. However, to date, no published studies have utilised a murine cell line to investigate this process. In the present study, we aimed to determine whether the mouse VSMC line MOVAS-1 can calcify in vitro. We established that the calcification of MOVAS-1 cells can be induced in the presence of calcifying medium (containing β-glycerophosphate and ascorbic acid), as detected by Alizarin Red and von Kossa staining, and quantification of calcium deposition and alkaline phosphatase activity. We also showed that the time course of MOVAS-1 calcification is comparable to that of the primary murine aortic VSMCs, establishing the MOVAS-1 cells as a feasible and relevant model. Significant increases in the mRNA expression profile of key genes associated with vascular calcification (Ocn, Akp2 and PiT-1) were observed in MOVAS-1 cells cultured under calcifying conditions, with similar changes in expression in murine aortic VSMCs. Furthermore, a significant reduction in calcification was observed in MOVAS-1 cells following treatment with levamisole and etidronate, known inhibitors of calcification. In conclusion, we demonstrated that the MOVAS-1 line is a reliable, convenient and economical system in which to investigate vascular calcification in vitro, and will make a useful contribution to increasing our understanding of this pathological process.
血管钙化在许多疾病中具有严重的临床后果，包括糖尿病、动脉粥样硬化和终末期肾病。原代小鼠、人源和牛源血管平滑肌细胞（VSMCs）的体外钙化常用于研究血管钙化的机制。然而，迄今为止，尚无研究利用小鼠细胞系来研究这一过程。本研究旨在确定小鼠 VSMC 系 MOVAS-1 是否可以在体外发生钙化。我们发现，在钙化培养基（含β-甘油磷酸和抗坏血酸）存在的情况下，MOVAS-1 细胞可以发生钙化，通过茜素红和 von Kossa 染色以及钙沉积和碱性磷酸酶活性的定量检测可以证实这一点。我们还表明，MOVAS-1 钙化的时间进程与原代小鼠主动脉 VSMCs 相似，这确立了 MOVAS-1 细胞作为一种可行且相关的模型。在钙化条件下培养的 MOVAS-1 细胞中，与血管钙化相关的关键基因（Ocn、Akp2 和 PiT-1）的 mRNA 表达谱显著增加，而在小鼠主动脉 VSMCs 中也观察到类似的表达变化。此外，在用莱米诺肽和依替膦酸治疗后，MOVAS-1 细胞中的钙化明显减少，莱米诺肽和依替膦酸是已知的钙化抑制剂。总之，我们证明了 MOVAS-1 系是一种可靠、方便且经济的体外研究血管钙化的系统，将有助于增加我们对这一病理过程的理解。
