Nicotine induces upregulated expression of beta defensin-2 via the p38MAPK pathway in the HaCaT human keratinocyte cell line
Nicotine induces upregulated expression of beta defensin-2 via the p38MAPK pathway in the HaCaT human keratinocyte cell line
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DOI：
10.1007/s00795-010-0493-4
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发表时间：
2010-12-01
影响因子：
1.8
通讯作者：
Abiko, Yoshihiro
中科院分区：
文献类型：
--
作者：
Nakamura, Sumiko;Saitoh, Masato;Abiko, Yoshihiro
人β-防御素（humanbeta-defensins，hBD）是一组抗菌肽，参与口腔上皮的保护屏障。尼古丁诱发牙周及口腔上皮疾病。本研究的目的是研究尼古丁对角质形成细胞hBD-2表达模式的影响。将HaCaT细胞（角质形成细胞系）与8、15、30或80 μ M尼古丁孵育24小时。通过RT-PCR、qRTPCR和ELISA检测hBD-2的表达。在一系列实验中，用细胞内途径抑制剂（p38 MAP激酶、NF-κ B、JNK、MAPK-ERK）和烟碱乙酰胆碱受体（nAChR）抑制剂处理细胞。使用Student t检验分析数据。qRT-PCR结果显示，在30和80 μ M尼古丁浓度下，hBD-2 mRNA的表达水平显著高于对照组（P < 0.05）。80 μ M细胞提取物含有比对照显著更高的hBD-2肽水平（P < 0.05）。p38 MAP激酶抑制剂可阻断尼古丁对hBD-2表达的上调作用。两种nAChR抑制剂也消除了尼古丁对hBD-2的上调。目前的研究表明，尼古丁通过p38 MAP激酶途径在角质形成细胞中引起hBD-2的表达上调。
Human beta-defensins (hBDs), a group of antimicrobial peptides, are involved in the protective barrier of the oral epithelium. Nicotine induces periodontal and oral epithelial diseases. The purpose of the present study was to investigate the effect of nicotine on the expression pattern of hBD-2 in keratinocytes. HaCaT cells, a keratinocyte cell line, were incubated with 8, 15, 30, or 80 mu M nicotine for 24 h. Expression of hBD-2 was observed by RT-PCR, qRTPCR, and ELISA assay. The cells were treated with inhibitors for intracellular pathways (p38MAP kinase, NF-kappa B, JNK, MAPK-ERK) and with nicotinic acetylcholine receptor (nAChR) inhibitors in a series of experiments. Data were analyzed using Student's t test. qRT-PCR revealed that the expression level of hBD-2 mRNA was significantly higher at 30 and 80 mu M nicotine than the control without nicotine (P < 0.05). The 80 mu M cell extraction contained significantly higher hBD-2 peptide levels than the control (P < 0.05). The p38MAP kinase inhibitor abolished the upregulated expression of hBD-2 by nicotine. Both nAChR inhibitors also abolished the upregulation of hBD-2 by nicotine. The present study demonstrated that nicotine causes upregulated expression of hBD-2 via the p38MAP kinase pathway in keratinocytes.