烟草对人牙龈成纤维细胞增殖及纤维连接蛋白表达的影响

目的:探讨烟草浸提液(smokeless tobacco,ST)对人牙龈成纤维细胞(human gingival fibroblasts,HGFs)在纯钛片上附着及增殖的影响及机制.方法:原代HGFs与不同浓度ST共同培养2h和2、4、6、8d;采用CCK-8法测定细胞黏附及增殖;采用双抗夹心酶联免疫吸附法检测不同时间培养上清液中纤维连接蛋白(fibroneetin,FN)的含量.采用SPSS19.0软件包对数据进行统计学分析.结果:HGFs在钛片上的黏附率随ST浓度升高呈下降趋势,5.0 g/L和10g/L的ST组黏附率分别为(34.316±7.725)％和(25.478±10.651)％,显著低于对照组(100％,P＜0.01).细胞在钛片上增殖至第2天和第4天,对照组显著大于2.5、5.0、10g/L ST组(P＜0.05);第6天和第8天时,对照组均显著大于0.625～10 g/L各ST组(P＜0.05).ST与HGFs培养到第8天,0.625、1.25、2.5 g/L的ST组FN浓度分别为(69.352±31.640)、(23.595±8.625)和(7.292±2.865) ng/mL,显著低于对照组(142.188±28.126)ng/mL (P＜0.05).结论:ST能显著抑制HGFs在钛片上的黏附和增殖,其机制可能与细胞产生FN减少有关.

Effect of tobacco extracts on human gingival fibroblasts' proliferation and fibronectin expression

PURPOSE:To investigate the influence and mechanism of smokeless tobacco (ST) extracts on proliferation and fibronectin expression of human gingival fibroblasts (HGFs) adhered to titanium plates.METHODS:The cultured primary HGFs with ST of different concentrations for a period of 2h and 2,4,6,8 d was separately conducted;the cell proliferation and the adhesion was assayed using CCK-8 method;enzyme-linked immunosorbent assay was used to determine the content of supernatant fibronectin (FN) at different times.The data was analyzed with SPSS 19.0 software package.RESULTS:With the increase of ST concentration,the cell adherence rate decreased accordingly.With ST concentration of 5.0 g/L and 10 g/L,the adhesion rate was (34.316±7.725)％ and (25.478±10.651)％,respectively,which was significantly lower than that of the control group (100％) (P＜0.01).Cell proliferation on the titanium plate started at 2 d and 4 d and the cell abundance of the control group was significantly greater than that of ST group (P＜0.05) with concentrations of 2.5,5.0,10 g/L;The cell abundance of the control group was significantly greater than that of ST group (P＜0.05) with concentrations of 0.625-10 g/L on the 6th and 8th day.In 0.625,1.25,2.5 g/L ST group,FN concentration was (69.352±31.640),(23.595±8.625) and (7.292±2.865) ng/mL,respectively,which was significantly lower than that of the control group (142.188 ± 28.126) ng/mL (P＜0.05).CONCLUSIONS:ST can significantly inhibit the proliferation and adhesion of HGFs on pure titanium surface,and the mechanism may be related to decrease of FN secreted by HGFs.