›› 2018, Vol. 38 ›› Issue (4): 314-319.

• 基础研究 • 上一篇    下一篇

透明质酸对尼古丁作用下人牙周膜成纤维细胞增殖影响研究

化萌萌1,王亚宇2,郭泾3   

  1. 1. 山东大学齐鲁医院
    2. 中国人民解放军306医院
    3. 山东大学口腔医院
  • 收稿日期:2017-03-10 修回日期:2017-04-26 出版日期:2018-04-28 发布日期:2018-05-03
  • 通讯作者: 郭泾 E-mail:guojing@sdu.edu.cn

The protective effects of hyaluronic acid against nicotine in human periodontal ligament fibroblasts

  • Received:2017-03-10 Revised:2017-04-26 Online:2018-04-28 Published:2018-05-03

摘要: 目的 观察不同浓度的尼古丁对人牙周膜成纤维细胞(HPDLFs)增殖的影响,并探讨透明质酸(HA)对尼古丁作用下对HPDLFs的保护作用。 方法 体外成功培养的HPDLFs,按不同的处理分为四组:(1)单纯细胞组(对照组);(2)尼古丁组:浓度分别为10μg/ml,100μg/ml,250μg/ml,500μg/ml,1000μg/ml的尼古丁;(3)HA组:浓度为0.1%和0.2%的HA;(4)尼古丁组+HA组共同培养(总10子组)。观察四组细胞增殖率,细胞碱性磷酸酶( ALP)的活性,Runt相关转录因-2(Runx2)、胶原蛋白Ⅰ(ColⅠ)、骨桥蛋白(OPN)、骨钙蛋白(OCN)的mRNA的水平,细胞钙矿化能力。结果 1、与对照组相比,各浓度尼古丁组均可以抑制HPDLFs的增殖率、ALP活性,Runx2 ,ColⅠ、OPN、OCN的mRNA水平,未见钙化矿化结节。2、与对照组相比,HA组均可以促进HPDLFs的增殖率、ALP活性和Runx2 ,ColⅠ、OPN、OCN的mRNA水平并观察到了钙化矿化结节。3、与尼古丁组相比,HA+尼古丁组对HPDLFs的增殖率和ALP活性起到促进作用,对Runx2 ,ColⅠ、OPN、OCN的mRNA的表达水平有提高并均观察到钙化矿化结节;各组差异具有统计学意义(P < 0.05 ) 结论 HA能够介导HPDLFs增殖,增加ALP活性和矿化组织相关蛋白,包括Runx2,ColⅠ,OPN和OCN等,并且抵抗尼古丁对HPDLFs的毒性作用,从而起到保护作用。

关键词: HPDLFs, HA, 尼古丁, 细胞增殖

Abstract: Objective To investigate the effects of hyaluronic acid (HA) against nicotine in human periodontal ligament fibroblasts (HPDLFs), and to explore the molecular mechanism underlying nicotine-induced cytotoxicity Methods This experiments using in vitro cultured human periodontal ligament fibroblasts (HPDLFs), divided into four groups: (1) normal group: cultured in DMEM medium continue; (2) the nicotine groups: add DMEM culture group concentration respectively (10 mu g/ml, 100 mu g/ml, 250 mu g/ml, 500 mu g/ml, 1000 mu g/ml) of nicotine. (3) hyaluronic acid groups: join in DMEM hyaluronic acid concentration was 0.1% and 0.2%; (4) the nicotine + hyaluronic acid groups: add concentration of 0.1% and 0.2% in DMEM HA and different concentrations of nicotine jointly develop 10 subgroups (total). Four groups were determined by MTT method (thiazole blue tetrazoliumbromide) cells, hence application kit of alkaline phosphatase (ALP) , Runt related transcription for - 2 (Runx2), collagen Ⅰ (Col Ⅰ) and osteopontin (OPN), osteocalcin (OCN) mRNA level, cell calcium mineralization ability .Results 1, compared with the control group, the concentration of nicotine can inhibit the proliferation activity of PDLFs and alkaline phosphatase activity, pure nicotine to join the group of Runx2, type I collagen, OPN and OCN had significantly lower levels of mRNA expression. Pure nicotine treatment group did not see calcification mineralized nodules.2, compared with the control group, respectively (0.1% and 0.2%) of hyaluronic acid on the periodontal ligament fibroblasts have obvious promoting effect, both kinds of concentration of hyaluronic acid can promote the proliferation activity of HPDLFs and alkaline phosphatase activity, and promote the rise of Runx2, type I collagen, the level of mRNA expression of OPN and OCN ,and obsered in mineralized nodules.3, compared with the pure nicotine effect group, hyaluronic acid + nicotine group of HPDLFs proliferation activity and alkaline phosphatase activities play a role, the Runx2, type I collagen, OPN and OCN mRNA expression level has improved, and observed calcification mineralized nodules. Conclusion HA can increase the activity of ALP and mineralized tissue-associated proteins, including Runx2, Col I, OPN and OCN, etc and can resist the toxic effect of nicotine on HPDLFs, and thus play a protective role.

Key words: HPDLFs, hyaluronic acid, nicotine, cell proliferation

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