牙龈卟啉单胞菌对血管平滑肌细胞钙化作用的研究  

Calcification of Vascular Smooth Muscle CellInduced by Porphyromonasgingivalis

作　　者：唐路[1] 薛栋[1] 白雨豪 王鹏程[1] 周维 李文婷[1] 董岩[1] 曾红燕[1] 赵颖[1] 

机构地区：[1]首都医科大学宣武医院口腔科,北京100053

出　　处：《口腔医学研究》2018年第2期130-133,共4页Journal of Oral Science Research

基　　金：首都卫生发展科研专项基金(编号:2016-4-2014)

摘　　要：目的:探讨牙龈卟啉单胞菌(Porphyromonasgingivalis,P.gingivalis)对小鼠血管平滑肌细胞(vascular smooth muscle cell,VSMC)钙化的影响。方法:选取C57BL/6小鼠,取主动脉体外培养VSMC,通过细胞免疫荧光染色,用α-smooth muscle actin(α-SMA)特异性抗体对VSMC鉴定。热灭活P.gingivalis以不同的感染倍数(multiplicity of infection,MOI)干预VSMC,分别在12、24、48、72h检查VSMC的细胞活性,在21天时检测VSMC钙化表现及钙含量。并且将P.gingivalis与小鼠主动脉共培养,观察P.gingivalis对小鼠主动脉的钙化作用。结果:特异性抗体α-SMA在细胞中荧光染色阳性,证明体外培养为VSMC。P.gingivalis(MOI:10,100)刺激VSMC在24和48h,细胞活性明显增强,在72h,所有感染倍数下(MOI:1,10,100)细胞活性较对照组均明显增强(P<0.05)。培养至21d时,P.gingivalis诱导VSMC出现明显钙化,钙含量与MOI成正相关(P<0.05)。P.gingivalis与小鼠主动脉培养14d,见主动脉壁出现明显钙化沉积。结论:P.gingivalis可诱导VSMC细胞活性增强及钙化,并且可直接引起主动脉壁钙化沉积。Objective：To explore the effects of porphyromonasgingivalis（P.gingivalis）on calcification of vascular smooth muscle cell（VSMC）.Methods：The primary culture of VSMC was isolated from the aortas of C57 BL/6 mice.VSMC was infected with heat-killed P.gingivalis at different MOI.The viability of VSMC was examined after 12,24,48 and 72 hours respectively.The calcification and calcium of VSMC were detected after 21 days.Then,P.gingivalis was co-cultured with aorta of mice to investigate the calcification of aorta.Results：VSMC was identified by immunofluorescence staining with specific antibodyα-SMA positive.The viability of VSMC stimulated by P.gingivalis（MOI：10,100）was significantly increased after 24 and 48 hours.All MOI groups enhanced cell viability after72 hours（P0.05）.The calcium content was positively correlated with P.gingivalis MOI（P0.05）.Then,the calcification of aorta induced by P.gingivalis was observed.Conclusion：VSMC viability and calcification areinduced by P.gingivalis,which also results in aortic wall calcification.

关 键 词：血管钙化 牙龈卟啉单胞菌 血管平滑肌细胞 牙周炎 动脉粥样硬化 

分 类 号：R781.4[医药卫生—口腔医学]