牙龈卟啉单胞菌脂多糖对共培养体系中人脐动脉平滑肌细胞增殖及迁移能力的影响  

Effects of Porphyromonas gingivalis lipopolysaccharide on the proliferation and migration of human umbilical artery smooth muscle cells under co-culture conditions

作　　者：李霞 胡琳琳 何权敏 高丽 葛颂 Li Xia;Hu Linlin;He Quanmin;Gao Li;Ge Song(Department of Periodontology,Stomatology Hospital of Zunyi Medical University,Zunyi 563000,China;General Department of Guiyang Stomatological Hospital,Guiyang 550002,China)

机构地区：[1]遵义医科大学附属口腔医院牙周科,563000 [2]贵阳市口腔医院综合科,550002

出　　处：《中华口腔医学杂志》2021年第6期549-556,共8页Chinese Journal of Stomatology

基　　金：国家自然科学基金(81260168,30760271)。

摘　　要：目的研究Ⅰ、ⅣfimA型牙龈卟啉单胞菌脂多糖(Porphyromonas gingivalis lipopolysaccharide,Pg-LPS)对共培养条件下人脐动脉平滑肌细胞(human umbilical artery smooth muscle cell,HUASMC)增殖和迁移能力的影响,探讨牙周炎与动脉粥样硬化(atherosclerosis,As)相关的生物学基础和机制。方法厌氧培养Ⅰ、ⅣfimA型Pg,分别提取、纯化并鉴定两型Pg-LPS;体外原代培养人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)和HUASMC,并采用鼠尾Ⅰ型胶原建立HUVEC-HUASMC共培养细胞模型;实验分为T1组(质量浓度为0.5、1.0、2.0、5.0、10.0 mg/L的ⅠfimA型Pg-LPS刺激共培养细胞)和T2组(质量浓度为0.5、1.0、2.0、5.0、10.0 mg/L的ⅣfimA型Pg-LPS刺激共培养细胞),以及阴性对照组(不加LPS组);细胞计数试剂盒(cell counting kit-8,CCK-8)法检测HUASMC的增殖能力,Transwell迁移小室观察HUASMC的迁移能力。比较各组不同质量浓度Pg-LPS刺激共培养细胞2、8、24及48 h后,HUASMC的增殖及迁移能力的变化。结果共培养细胞在Ⅰ、ⅣfimA型Pg-LPS的作用下,在24及48 h时,两型Pg-LPS的各质量浓度组中,HUASMC的A值均较阴性对照组显著上调(P<0.05);在48 h时5.0、10.0 mg/LⅣfimA型Pg-LPS刺激共培养细胞后,HUASMC的A值(分别为1.386±0.044、1.455±0.058)均显著高于相同浓度ⅠfimA型Pg-LPS刺激共培养中HUASMC的A值(分别为1.168±0.064、1.204±0.088)(P<0.05);此外,在48 h时,5.0、10.0 mg/LⅣfimA型Pg-LPS刺激共培养细胞后,HUASMC的A值均显著高于0.5、1.0 mg/LⅣfimA型Pg-LPS刺激共培养中HUASMC的A值(分别为1.170±0.082、1.239±0.089)(P<0.05)。HUASMC的迁移结果显示,在8、24及48 h时,Ⅰ、ⅣfimA型Pg-LPS各质量浓度组中,HUASMC的迁移数量均较同组内2 h时相同Pg-LPS质量浓度下HUASMC迁移数量显著上调(P<0.05);在48 h时,除10.0 mg/LⅣfimA型Pg-LPS外,其余各Pg-LPS质量浓度组HUASMC的迁移数量均较同组内24 h时相同Pg-LPS质量浓度下HUASMC迁移数量显Objective To investigate the effects of Porphyromonas gingivalis lipopolysaccharide(Pg-LPS)of typeⅠandⅣfimA on the proliferation and migration of human umbilical artery smooth muscle cells(HUASMC)under co-culture conditions and to explore the biological basis and possible mechanisms of the relationship between periodontitis and atherosclerosis(As).Methods TypeⅠandⅣfimA Pg were anaerobically cultured and the Pg-LPS was extracted,purified and identified.Human umbilical vein endothelial cells(HUVEC)and HUASMC were cultured in vitro and the HUVEC-HUASMC co-cultured cell model was established using rat tail typeⅠcollagen.The experiment was divided into three groups:group T1(co-cultured cells were stimulated with typeⅠfimA Pg-LPS at the mass concentrations of 0.5,1.0,2.0,5.0,10.0 mg/L),group T2(co-cultured cells were stimulated with typeⅣfimA Pg-LPS at the mass concentrations of 0.5,1.0,2.0,5.0,10.0 mg/L),and negative control group(without LPS).Cell counting kit-8(CCK-8)was used to detect the proliferation ability of HUASMC and the migration ability of HUASMC was observed by using the Transwell migration chamber.Comparasons of the changes in the proliferation and migration ability of HUASMC after 2,8,24 and 48 h of Pg-LPS stimulated co-cultured cells with various mass concentrations of Pg-LPS were conducted.Results For the co-cultured cells under the action of typeⅠandⅣfimA Pg-LPS,at 24 and 48 h,in each mass concentration group of the two types of Pg-LPS(0.5,1.0,2.0,5.0,10.0 mg/L),HUASMC′s A values were significantly up-regulated compared to the negative control group(P<0.05)and for the co-cultured cells after the stimulation of typeⅣfimA Pg-LPS at concentrations of 5 and 10 mg/L at 48 h,the A values(1.386±0.044,1.455±0.058)of HUASMC were significantly higher than that of HUASMC(1.168±0.064,1.204±0.088)in the same concentrations of typeⅠfimA Pg-LPS(P<0.05).In addition,the A values of HUASMC in stimulated co-cultured cells under concentrations of 5.0 and 10.0 mg/L of typeⅣfimA Pg-LPS at 48

关 键 词：紫单胞菌 龈 脂多糖类 人脐动脉平滑肌细胞 人脐静脉内皮细胞 动脉粥样硬化 细胞增殖 细胞迁移 

分 类 号：R781.42[医药卫生—口腔医学]