Nicotine-induced expression of low-density lipoprotein receptor in oral epithelial cells.
Nicotine-induced expression of low-density lipoprotein receptor in oral epithelial cells.
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DOI：
10.1371/journal.pone.0082563
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Ito K
中科院分区：
文献类型：
--
作者：
Ito S;Gojoubori T;Tsunoda K;Yamaguchi Y;Asano M;Goke E;Koshi R;Sugano N;Yoshinuma N;Komiyama K;Ito K
尼古丁的使用是心血管疾病和牙周病发生的最重要的危险因素之一。许多报告表明，脂质代谢紊乱可能对这两种疾病的发展有贡献。尽管有这些观察结果，但人们对吸烟与这些疾病的发展之间的关系知之甚少。我们之前的微阵列数据显示，尼古丁诱导口腔上皮细胞（OECs）中的低密度脂蛋白受体（LDLR）表达。本研究的目的是证实尼古丁介导的LDLR诱导，并阐明导致oec中LDLR表达增强的信号机制。实时荧光定量PCR检测LDLR和烟碱乙酰胆碱受体亚基表达。免疫荧光染色证实了LDLR的产生。通过nachr特异性抑制剂α-班加罗毒素（α-BTX）对细胞进行预孵育，检测nachr介导的LDLR诱导。通过荧光素酶测定、米霉素预孵育或小干扰RNA （siRNA）转染检测转录因子特异性蛋白1 （Sp1）的功能重要性。采用电泳迁移率转移法（EMSA）、链亲和素-琼脂糖沉淀法和western blotting验证Sp1与LDLR 5 ' -未翻译区R3区的特异性结合。结果证实尼古丁在转录水平诱导了LDLR的表达。尼古丁被nAChR感知，信号被Sp1转导到LDLR基因的R3区。通过对吸烟患者的牙龈组织进行免疫荧光染色，进一步证实了牙龈上皮细胞中LDLR的增加。综上所述，结果表明尼古丁可能通过诱导oec中的LDLR从而干扰脂质代谢而促进心血管和牙周病的发展。
Nicotine use is one of the most important risk factors for the development of cardiovascular and periodontal diseases. Numerous reports have suggested the possible contribution of disturbed lipid metabolism for the development of both disease groups. Despite these observations, little is known about the relationship between tobacco smoking and the development of these diseases. Our previous microarray data revealed that nicotine induced low-density lipoprotein receptor (LDLR) expression in oral epithelial cells (OECs). The aim of the present study was to confirm nicotine-mediated LDLR induction and to elucidate the signaling mechanisms leading to the augmented expression of LDLR in OECs. LDLR and nicotinic acetylcholine receptor (nAChR) subunit expression was detected by real-time PCR. The production of LDLR was demonstrated by immunofluorescence staining. nAChR-mediated LDLR induction was examined by pre-incubation of the cells with its specific inhibitor, α-bungarotoxin (α-BTX). The functional importance of transcription factor specific protein 1 (Sp1) was examined by luciferase assay, mithramycin pre-incubation or by small interfering RNA (siRNA) transfection. The specific binding of Sp1 to R3 region of LDLR 5’-untranslated region was demonstrated with electrophoretic mobility shift assay (EMSA) and streptavidin-agarose precipitation assay followed by western blotting. The results confirmed that nicotine induced LDLR expression at the transcriptional level. Nicotine was sensed by nAChR and the signal was transduced by Sp1 which bound to the R3 region of LDLR gene. Augmented production of LDLR in the gingival epithelial cells was further demonstrated by immunofluorescence staining using the gingival tissues obtained from the smoking patients. Taken together, the results suggested that nicotine might contribute to the development of both cardiovascular and periodontal diseases by inducing the LDLR in OECs thereby disturbing lipid metabolism.
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影响因子：
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通讯作者：
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影响因子：
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