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  

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

作　　者：化萌萌 王亚宇 郭泾[1,2] HUA Mengmeng;WANG Yayu;GUO Jing(Department of Orthodontics, School of Stomatology, Shandong University, Ji＇nan 250012, China)

机构地区：[1]山东大学口腔医学院正畸科,山东济南250012 [2]山东省口腔生物医学重点实验室,山东济南250012 [3]山东大学齐鲁医院口腔科,山东济南250012 [4]解放军306医院全军口腔疾病诊疗中心,北京100101

出　　处：《口腔医学》2018年第4期314-319,共6页Stomatology

基　　金：山东省自然科学基金重点项目(ZR2014HZ001)

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4组:(1)单纯细胞组(对照组);(2)尼古丁组:浓度分别为10、100、250、500、1 000μg/m L;(3)HA组:浓度为0.1%和0.2%;(4)尼古丁组+HA组(总10子组)。观察4组细胞增殖率,ALP的活性,Runx2、ColⅠ、OPN、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。Objective To investigate the effects of hyaluronic acid against nicotine on human periodontal ligament fibroblasts,and to explore the molecular mechanism underlying nicotine-induced cytotoxicity. Methods In vitro cultured human periodontal ligament fibroblasts（ HPDLFs） were divided into four groups according to the treatment：（1）normal group： cultured in DMEM medium continue;（2）the nicotine groups： concentrations of nicotine were added respectively（10 μg/m L,100 μg/m L,250 μg/m L,500 μg/m L,1 000μg/m L）.（3）hyaluronic acid groups： in DMEM hyaluronic acid concentrations were 0.1% and 0.2%;（4）the nicotine ＋ hyaluronic acid groups： concentrations of 0.1% and 0. 2% in DMEM HA and different concentrations of nicotine jointly developed into 10 subgroups（total）. Four groups were determined by MTT method（thiazole blue tetrazoliumbromide）. The cell proliferation rate,activity of ALP,the mRNA levels of Runx2,Col Ⅰ,OPN and OCN,and cell calcium mineralization ability were observed. Results（1）Compared with the control group,the concentration of nicotine could inhibit the proliferation activity of PDLFs,alkaline phosphatase activity,and levels of mRNA expression of Runx2,ColⅠ,OPN and OCN. Calcification mineralized nodules were not seen in pure nicotine treatment group.（2）Compared with the control group,concentrations（ 0.1% and 0.2%） of hyaluronic acid had an obvious promoting effect on the periodontal ligament fibroblasts,both kinds of concentrations of hyaluronic acid could promote the proliferation activity of HPDLFs and alkaline phosphatase activity,and promote the rise of Runx2,ColⅠ,the level of mRNA expression of OPN and OCN,and mineralized nodules were observed.（3）Compared with the pure nicotine effect group,hyaluronic acid ＋ nicotine group played a role in promoting HPDLFs proliferation activity and alkaline phosphatase activities,the mRNA expression level of Runx2,ColⅠ,OPN and OCN improved,and calcification mineralized nodules were observed. Concl

关 键 词：人牙周膜成纤维细胞 HA 尼古丁 细胞增殖 

分 类 号：R781.4[医药卫生—口腔医学]