机械应力诱导HMGB1通过金属基质蛋白酶促进人牙周膜细胞的迁移
HMGB1 induced by mechanical stress promotes the migration of human periodontal ligament cells through regulating matrix metalloproteinases
余小琴 1高锦瑜 2刘华1
作者信息
- 1. 安康市人民医院口腔科,陕西安康725000
- 2. 延安大学附属医院口腔医院正畸科,陕西延安716000
- 折叠
摘要
目的:探究高迁移率蛋白1(HMGB1)在机械应力诱导的人牙周膜细胞(hPDLCs)迁移中的作用及机制.方法:用Real-time PCR和Western blotting方法检测机械应力(10％形变量、频率0.5 Hz)加载不同时间(0、6、12、24、48 h)对hPDLCs中HMGB1和金属基质蛋白酶2、9(MMP2/9)表达的影响.然后,使用siR-NA干扰技术下调HMGB1,并分别用Transwell检测hPDLCs迁移力、Real-time PCR检测MMPs表达、Westernblotting检测ERK1/2和p-ERK1/2的表达水平.另外,分别使用1μmol/L MMPs抑制剂Batimastat、50 nmol/LERK1/2信号通路抑制剂GDC-0994预处理hPDLCs 2 h,然后再加载机械应力24h,并分别检测细胞迁移或MMPs的表达.结果:hPDLCs经机械应力加载后,其HMGB1、MMP2、MMP9 mRNA和蛋白的表达水平均随着加载时间而逐渐增加,24 h时达到峰值,48 h时呈下降趋势;下调HMGB1能够抑制机械应力诱导的hPDLCs迁移及细胞中MMP2、MMP9和p-ERK1/2的表达;Batimastat能够抑制机械应力诱导的细胞迁移;GDC-0994能够抑制机械应力诱导的MMP2和MMP9的表达.结论:HMGB1介导机械应力诱导的hPDLCs迁移,可能是通过激活ERK1/2信号通路、上调MMPs的表达而发挥作用的.
Abstract
AIM:To study the role and mechanism of high mobility group box protein 1 (HMGB1) in mechanical stress induced the migration of human periodontal ligament cells (hPDLCs).METHODS:The expression of HMGB1 and matrix metalloproteinase 2/9 (MMP2/9) was assayed by real-time PCR and western blotting when cells were cultured with mechanism stress with 10％ deformation,and 0.5 Hz sinusoidal wave for 0,6,12,24 and 48 h respectively.Then,HMGB1 expression was downregulated by siRNA,the migration of hPDLCs was examined by transwell,MMPs expression was analyzed by real-time PCR,and the levels of ERK1/2 and p-ERK1/2 were determined by western blotting.Additionally,hPDLCs were pretreated with 1 μmol/L of Batimastat (a MMPs inhibitor) and 50 nmol/L of GDC-0994 (an ERK1/2 inhibitor) respectively for 2 h and then treated with mechanism stress for 24 h,the cell migration and MMPs expression was detected,respectively.RESULTS:The mRNA and protein expression of HMGB1,MMP2 and MMP9 were all increased at 6,12 and 24 h in a time-dependent manner,while reduced at 48 h.Downregulation of HMGB1 inhibited mechanical stress-induced the migration of hPDLCs and the expression of MMP2,MMP9 and p-ERK1/2.Batimastat suppressed hPDLCs migration.The expression of MMP2 and MMP9 induced by mechanical stress was dampened by GDC-0994.CONCLUSION:HMGB1 mediates mechanical stress-induced the migration of hPDLCs,which may be through activating ERK1/2 signaling and upregulating MMPs expression.
关键词
HMGB1/机械应力/人牙周膜细胞/迁移/金属基质蛋白酶Key words
HMGB1/mechanical stress/human periodontal ligament cells/migration/matrix metalloproteinase引用本文复制引用
出版年
2017
NETL
NSTL
维普
万方数据