烟草浸提液对成骨细胞生物学性能的影响  

Smokeless Tobacco Extract Affects Biological Properties of the Pre-osteoblast Cell Line MC3T3-E1

作　　者：薛鹏飞[1] 侯玉东[1] 张云涛[2] 潘晶晶[3] 孙鑫[1] 孙龙[1] 

机构地区：[1]滨州医学院口腔医学院,烟台264003 [2]滨州医学院附属医院口腔修复科,滨州256603 [3]潍坊医学院医学影像学系,潍坊261053

出　　处：《四川大学学报（医学版）》2014年第6期908-912,共5页Journal of Sichuan University(Medical Sciences)

基　　金：山东省医药卫生科技发展计划项目(No.2013WS0299)资助

摘　　要：目的研究不同浓度烟草浸提液（smokeless tobacco extract,STE）对成骨细胞生物学性能的影响,探讨吸烟影响种植体骨结合的病理机制。方法以0（对照组）、0.01、0.1、1、5、10g/L的STE作用于小鼠颅顶前成骨细胞亚克隆14（MC3T3-E1Subclone 14,MC3T3）,MTT法检测在STE作用1、3、5、7d后的增殖情况,罗丹明及DAPI染色后以激光扫描共聚焦显微镜（confocal laser scanning microscope）观察MC3T3细胞骨架（cytoskeleton）在STE作用24h后的形态变化,荧光实时定量PCR（quantitative real-time PCR,RT-qPCR）检测白细胞介素-6（interleukin-6,IL-6）和核心结合因子α1（Cbfα1）mRNA在STE作用48h后的表达。结果 MTT试验显示0.01～10g/L的STE可抑制MC3T3增殖（P〈0.05）,5～10g/L的STE可随时间延长抑制MC3T3增殖（P〈0.05）。细胞骨架观察显示对照组细胞骨架呈网络状铺展,在STE刺激下微丝解聚,细胞骨架重排且随浓度升高作用更加明显。RT-qPCR结果显示5～10g/L的STE可促进MC3T3的IL-6mRNA表达（P〈0.05）,0.1～10g/L的STE可抑制MC3T3的Cbfα1mRNA表达（P〈0.05）,随STE浓度升高,作用更加明显。结论烟草可抑制成骨细胞增殖,影响细胞骨架结构,增高MC3T3细胞IL-6mRNA的表达和降低其Cbfα1mRNA的表达,从而抑制成骨细胞的分化及附着,提示吸烟不利于骨结合。Objective To investigate the effects of smokeless tobacco extract（STE）on biological properties of osteoblast,and to identify possible pathological mechanisms of osseointegration.Methods MC3T3-E1Sub-clone14 cells were cultured in the presence of STE at 0（control group）,0.01,0.1,1,5,10g/L.The cell proliferation was measured by MTT assay 1d,3d,5d,and 7dafter exposure.The F-actin cytoskeleton of MC3T3 was stained with Rhodamine and DAPI,and then examined under a confocal laser scanning microscope 24 hafter exposure to STE.The mRNA expressions of interleukin-6（IL-6）and core-binding factorα1（Cbfα1）were quantified by realtime PCR（RT-qPCR）48hafter exposure to STE.Results The MTT assay showed that 0.01-10g/L STE inhibited MC3T3proliferation（P〈0.05）.Prolonged time enabled 5-10g/L STE to inhibit MC3T3proliferation（P〈0.05）.Network structure in F-actin cytoskeleton was demonstrated in the controls.In the cells exposed to STE,F-actin cytoskeleton started to change with disruptive structures.As the concentration of STE increased,the changes became more significant.STE increased the mRNA expression of IL-6at the concentration of 5g/L and10g/L（P〈0.05）,decreased the mRNA expression of Cbfα1at the concentration of 0.1-10g/L（P〈0.05）.Conclusion Tobacco may inhibit osteoblast proliferation,destroy F-actin cytoskeleton structure,increase the mRNA expression of IL-6and decrease the mRNA expression of Cbfα1,and inhibit cell differentiation and adhesion accordingly.Smoking is a disadvantage to osseointegration.

关 键 词：烟草 增殖 细胞骨架 IL-6 核心结合因子Α1 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]