MicroRNAs that target Ca2+ transporters are involved in vascular smooth muscle cell calcification
MicroRNAs that target Ca2+ transporters are involved in vascular smooth muscle cell calcification
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DOI：
10.1038/labinvest.2012.85
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发表时间：
2012-09-01
影响因子：
5
通讯作者：
Muragaki, Yasuteru
中科院分区：
文献类型：
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作者：
Gui, Ting;Zhou, Gengyin;Muragaki, Yasuteru
MicroRNAs(MiRNAs)在血管钙化中的作用目前尚不清楚。为了研究miRNAs是如何参与血管平滑肌细胞(VSMC)钙化的，我们探讨了在体外和体内VSMC钙化过程中miRNAs的变化。Klotho纯合子突变小鼠(KL/KL)表现出血管钙化，并有钙处理的扰动。因此，我们假设VSMC中的钙扰动可能是由miRNAs介导的。利用miRNA阵列分析，我们证明了与野生型(WT)小鼠相比，3周龄KL/KL小鼠的主动脉中膜中miRNAs的表达发生了异常。KL/KL小鼠主动脉中膜miR-135a(STAR)、miR-762、miR-714和miR-712(STAR)的表达水平显著高于WT小鼠。我们使用实时定量逆转录聚合酶链式反应进一步证实这些miRNAs在KL/KL小鼠的主动脉中膜和高磷、高钙处理的培养的VSMC中增加。对miRNA数据库的搜索表明，钙外流蛋白NCX1、PMCA1和NCKX4经常出现在这些miRNAs的潜在靶标中。将miRNA模拟物导入培养的VSMC中，降低了每个潜在靶点的蛋白质水平。相反，miRNA抑制剂减少了磷酸盐和钙诱导的VSMC钙化。此外，这些抑制剂还能降低磷酸盐和钙离子处理后培养的VSMC内钙离子浓度。我们的结果提示，血管平滑肌细胞miR-135a(STAR)、miR-762、miR-714和miR-712(STAR)的表达增加可能通过破坏钙外流蛋白参与VSMC的钙化。实验室调查(2012年)92,1250-1259；doi：10.1038/Labinvest.2012.85；2012年6月11日在线发布
The role of microRNAs (miRNAs) in vascular calcification is currently unclear. To examine how miRNAs are involved in vascular smooth muscle cell (VSMC) calcification, we explored the alteration of miRNAs in VSMC calcification in vitro and in vivo. Klotho homozygous mutant mice (kl/kl) display vascular calcification and have perturbations of calcium handling. We therefore hypothesized that the calcium perturbations in VSMCs could be mediated by miRNAs. Using an miRNA array analysis, we demonstrated that miRNAs are aberrantly expressed in the aortic media of 3-week-old kl/kl mice compared with wild-type (WT) mice. The expression levels of miR-135a(star), miR-762, miR-714, and miR-712(star) in the aortic media of kl/kl mice were significantly higher than in WT mice. We used quantitative real-time reverse transcriptase polymerase chain reaction to further confirm that these miRNAs were increased in the aortic media of kl/kl mice and in cultured VSMCs treated with high phosphate and calcium. A search of the miRNA database indicated that the Ca2+ efflux proteins NCX1, PMCA1, and NCKX4 frequently appeared as potential targets of these miRNAs. The transfection of miRNA mimics into cultured VSMCs reduced the protein levels of each potential target. Conversely, miRNA inhibitors reduced phosphate and calcium-induced VSMC calcification. Furthermore, these inhibitors decreased the intracellular Ca2+ concentration in cultured VSMCs after treatment with phosphate and calcium. Our results suggest that increased expression of miR-135a(star), miR-762, miR-714, and miR-712(star) in VSMCs may be involved in VSMC calcification by disrupting Ca2+ efflux proteins. Laboratory Investigation (2012) 92, 1250-1259; doi:10.1038/labinvest.2012.85; published online 11 June 2012