摘要
目的通过观察牙龈卟啉单胞菌脂多糖(PgLPS)对人脐静脉内皮细胞(EA.hy926)的影响,探讨PgLPS在动脉粥样硬化(AS)的发生发展过程中可能发挥的作用。方法本文以EA.hy926细胞为研究对象,随机提取2~4代增殖旺盛的细胞进行实验(n=5),用不同浓度(0.01、0.1、1、10 mg/L)PgLPS刺激EA.hy926细胞,用细胞增殖检测试剂盒(CCK8)检测PgLPS对其细胞增殖影响,用透射电镜观察PgLPS对其细胞结构变化的影响,用酶联免疫吸附试验(ELISA)法检测细胞上清可溶型细胞间黏附分子1(sICAM-1),以及用Western-blot法检测细胞膜结合型细胞间黏附分子1(mICAM-1)的表达。结果当PgLPS浓度达到1 mg/L时,作用时间为24 h以上即可抑制EA.hy926细胞增殖(P<0.05);PgLPS浓度达到0.1 mg时透射电子显微镜可观察出现符合内皮细胞焦亡的改变;低浓度和高浓度PgLPS均能促进内皮细胞sICAM-1的表达增加(P<0.01),其中0.1 mg/L组sICAM-1表达水平最高,且上清液中的sICAM-1浓度随着PgLPS作用时间的延长而递增;Western-blot法检测结果显示PgLPS浓度达到1、10 mg/L时内皮细胞mICAM-1的相对表达量明显下降;在PgLPS浓度为10 mg/L时,不同的刺激时间作用下,各组mICAM-1的表达差异无统计学意义。结论PgLPS可以抑制人脐静脉内皮细胞增殖,使其发生焦亡的形态学改变,促进其sICAM-1的表达。
Objective To evaluate the potential role of porphyromonas gingivalis lipopolysaccharides(PgLPS)in the development of atherosclerosis.Methods Human umbilical vein endothelial cells(EA.hy926)were stimulated with PgLPS at the concentration of 0.01,0.1,1 and 10 mg/L PgLPS respectively.Proliferation assay kit(CCK8)was used to detect the effect of PgLPS on the proliferation of EA.hy926 cells.The effect of PgLPS on cellular structure was observed by transmission electron microscopy.The soluble intercellular adhesion molecule-1(sICAM-1)in the supernatant of EA.hy926 cells was detected by enzyme-linked immunosorbent assay(ELISA).The expression of membrane-bound intercellular adhesion molecule 1(mICAM-1)was detected by Western-blot.Results When concentration of PgLPS reached 1 mg/L,the proliferation of EA.hy926 cells were inhibited(P<0.05)after 24 hours.When concentration of PgLPS reached 0.1 mg/L,the pyroptosis were observed under transmission electron microscope.The expression levels of sICAM-1 at all concentration of PgLPS increased(P<0.01)with highest expression level at 0.1 mg/L.The concentration of sICAM-1 in supernatant increased over time under the stimulation of PgLPS.The expression of mICAM-1 decreased when the concentration of PgLPS reached to 1 and 10 mg/L.Once the concentration of PgLPS reached to 10 mg/L,the expression of mICAM-1 showed no statistical difference over time.Conclusion PgLPS can inhibit the proliferation of EA.hy926 cells and may induce pyroptosis.PgLPS can induce the expression of sICAM-1.
作者
吴贇
叶春玲
林李嵩
陈凌
林志坚
WU Yun;YE Chun-ling;LIN Li-song;CHEN Ling;LIN Zhi-jian(Stomatology Department,The First Affiliated Hospital of Fujian Medical University,Fuzhou Fujian 350005,China;不详)
出处
《中华高血压杂志》
CAS
CSCD
北大核心
2020年第5期420-425,共6页
Chinese Journal of Hypertension
基金
福建省自然科学基金资助项目(2017J05124)
中华口腔医学会口腔疾病与全身疾病关系研究项目(CSAY2015-09)。
关键词
牙龈卟啉单胞菌脂多糖
细胞间黏附分子-1
细胞焦亡
porphyromonas gingivalis lipopolysaccharides
intercellular adhesion molecule 1
pyroptosis
