α-klotho蛋白在人真皮微血管内皮细胞迁移和增殖中的作用。
Role of Klotho in migration and proliferation of human dermal microvascular endothelial cells.
作者信息
Markiewicz Margaret, Panneerselvam Kavin, Marks Natalia
机构信息
Division of Rheumatology and Immunology, Medical University of South Carolina, Charleston, SC 29425, United States.
Division of Rheumatology and Immunology, Medical University of South Carolina, Charleston, SC 29425, United States.
出版信息
Microvasc Res. 2016 Sep;107:76-82. doi: 10.1016/j.mvr.2016.05.005. Epub 2016 May 31.
PURPOSE
To examine the possible role of Klotho (Kl) in human microvasculature.
METHODS
The expression level of Kl in primary human dermal microvascular endothelial cells (HDMECs) and primary human dermal fibroblasts (HFb) was detected by real-time polymerase chain reaction amplification (qRT-PCR), Western blot analyses and immunohistochemistry. Migration of HDMECs and HFb was examined in monolayer wound healing "scratch assay" and Transwell assay. Proliferation of these cells was examined using Cell Proliferation BrdU incorporation assay.
RESULTS
Our results have shown that downregulation of Kl abrogated HDMECs migration after 48h. On the other hand, migration of HFb significantly increased after blocking Kl. Lack of Kl decreased expression of genes involved in the activation of endothelial cells and enhanced expression of genes involved in extracellular matrix remodeling and organization of connective tissue.
CONCLUSIONS
This study for the first time provides the evidence that Kl is expressed in HDMECs and HFb. Additionally, we have demonstrated that Kl is implicated in the process of angiogenesis of human dermal microvasculature.
目的
研究Klotho（Kl）在人体微血管系统中可能发挥的作用。
方法
通过实时聚合酶链反应扩增（qRT-PCR）、蛋白质免疫印迹分析和免疫组织化学检测人原代表皮微血管内皮细胞（HDMECs）和人原代表皮成纤维细胞（HFb）中Kl的表达水平。在单层伤口愈合“划痕试验”和Transwell试验中检测HDMECs和HFb的迁移情况。使用细胞增殖BrdU掺入试验检测这些细胞的增殖情况。
结果
我们的研究结果表明，Kl的下调在48小时后消除了HDMECs的迁移。另一方面，阻断Kl后HFb的迁移显著增加。缺乏Kl会降低参与内皮细胞激活的基因表达，并增强参与细胞外基质重塑和结缔组织组织化的基因表达。
结论
本研究首次提供了Kl在HDMECs和HFb中表达的证据。此外，我们证明了Kl与人原代表皮微血管的血管生成过程有关。
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