Antimicrobial peptide LL37 promotes vascular endothelial growth factor-A expression in human periodontal ligament cells.
Antimicrobial peptide LL37 promotes vascular endothelial growth factor-A expression in human periodontal ligament cells.
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DOI：
10.1111/j.1600-0765.2012.01524.x
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发表时间：
2013-04
影响因子：
3.5
通讯作者：
M. Kittaka;H. Shiba;M. Kajiya;K. Ouhara;K. Takeda;K. Kanbara;T. Fujita;H. Kawaguchi;H. Komatsuzawa;H. Kurihara
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文献类型：
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作者：
M. Kittaka;H. Shiba;M. Kajiya;K. Ouhara;K. Takeda;K. Kanbara;T. Fujita;H. Kawaguchi;H. Komatsuzawa;H. Kurihara
背景和目的 LL37 最初存在于先天免疫系统中，是一种强大的抗菌肽。 LL37 除了具有抗菌活性外，在各种细胞类型中还表现出多种生物功能，如迁移、细胞因子产生、细胞凋亡和血管生成。牙周膜 (PL) 细胞在牙周组织再生中发挥着关键作用。基于这些发现，我们假设LL37可以调节PL细胞功能以促进牙周组织的再生。为了证明这一假设，我们研究了 LL37 对人 PL (HPL) 细胞培养物中强效血管生成诱导剂血管内皮生长因子 (VEGF) 表达的影响，因为新血管形成对于组织再生的进展是不可或缺的。此外，我们研究了与 LL37 诱导的 VEGF 表达相关的信号级联反应。材料和方法 在存在或不存在 PD98059（一种 MEK-ERK 抑制剂）或 PDTC（一种 NF-κB 抑制剂）的情况下，用合成的 LL37 处理 HPL 细胞。通过实时聚合酶链反应分析和酶联免疫分析评估 VEGF 表达水平。通过蛋白质印迹法测定 ERK1/2 或 NF-κB p65 的磷酸化水平。结果 LL37 上调 HPL 细胞中 VEGF-A mRNA 和蛋白水平的表达，而 VEGF-B mRNA 表达不受影响。 ERK 和 NF-κB 抑制剂均明显消除了 HPL 细胞中 LL37 诱导的 VEGF-A 水平增加。重要的是，LL37 增加了 HPL 细胞中 ERK1/2 和 NF-κB p65 的磷酸化水平。结论 LL37 通过 ERK 和 NF-κB 信号级联诱导 HPL 细胞产生 VEGF-A，这可能导致血管生成，从而有助于牙周再生。
BACKGROUND AND OBJECTIVE LL37, originally found in the innate immune system, is a robust antimicrobial peptide. LL37 exhibits multiple bio-functions in various cell types, such as migration, cytokine production, apoptosis, and angiogenesis besides its antimicrobial activity Periodontal ligament (PL) cells play a pivotal role in periodontal tissue regeneration. Based on these findings, we hypothesized that LL37 can regulate PL cell function to promote regeneration of periodontal tissue. To prove this hypothesis, we investigated the effect of LL37 on the potent angiogenic inducer vascular endothelial growth factor (VEGF) expression in cultures of human PL (HPL) cells because neovascularization is indispensable for the progress of tissue regeneration. Moreover, we investigated the signaling cascade associated with LL37-induced VEGF expression. MATERIAL AND METHOD HPL cells were treated with synthesized LL37 in the presence or absence of PD98059, a MEK-ERK inhibitor, or PDTC, an NF-κB inhibitor. VEGF expression levels were assessed by real-time polymerase chain reaction analysis and an enzyme-linked immunoassay. Phosphorylation levels of ERK1/2 or NF-κB p65 were determined by Western blotting. RESULTS LL37 upregulated VEGF-A expression at the mRNA and protein levels in HPL cells, while VEGF-B mRNA expression was not affected. Both ERK and NF-κB inhibitors clearly abrogated the increase in VEGF-A levels induced by LL37 in HPL cells. Importantly, LL37 increased phosphorylated levels of ERK1/2 and NF-κB p65 in HPL cells. CONCLUSION LL37 induces VEGF-A production in HPL cells via ERK and NF-κB signaling cascades, which may result in angiogenesis, thereby contributing to periodontal regeneration.