Effects of Porphyromonas endodontalis lipopolysaccharides on the expression of matrix metalloproteinase-9 in mouse osteoblasts
10.3760/cma.j.issn.1002-0098.2017.08.010
- VernacularTitle: 牙髓卟啉单胞菌脂多糖对小鼠成骨细胞基质金属蛋白酶9表达的影响
- Author:
Xiaolin LI
1
;
Yaqiong YU
1
;
Lihong QIU
1
;
Di YANG
1
;
Xuemei WANG
1
;
Jingtao YU
1
Author Information
1. Department of Endodontics, School of Stomatology, China Medical University & Liaoning Institute of Dental Research, Shenyang 110002, China
- Publication Type:Journal Article
- Keywords:
Porphyromonas endodontalis;
Lipopolysaccharides;
Matrix metalloproteinase 9;
Osteoblasts;
Signaling pathway
- From:
Chinese Journal of Stomatology
2017;52(8):499-503
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To evaluate the effects of lipopolysaccharides (LPS) extracted from Porphyromonas endodontalis (Pe) on the expression of matrix metalloproteinase-9 (MMP-9) mRNA and protein as well as enzyme activity in MC3T3-E1 cells and the role of nuclear factor-κB (NF-κB) in the process, so as to investigate the expression of MMP-9 dependent signaling pathways in mouse osteoblasts induced by Pe LPS.
Methods:The experiment was conducted in 3 sessions: MC3T3-E1 cells were treated with various concentrations of Pe LPS (0-20 mg/L) and 10 mg/L Pe LPS for different time intervals (0-48 h). The expression of MMP-9 mRNA and protein were detected by real-time reverse transcription-PCR (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), while the enzyme activity was detected by gelatin zymography method. The expression of MMP-9 mRNA was also detected in 10 mg/L Pe LPS treated MC3T3-El cells after pretreated with specific NF-κB inhibitor BAY 11-7082 for l h. Statistical analysis was performed using one-way ANOVA and Dunnett t test with SPSS 13.0 software package.
Results:The levels of MMP-9 mRNA and protein increased significantly after the treatment with various concentrations of Pe LPS (0-20 mg/L), which indicated that Pe LPS induced osteoblasts to express MMP-9 in dose dependent manners. The expression of MMP-9 protein increased from (5 395±362) ng/L (blank control group) to (12 684±375) ng/L (20 mg/L group). Maximal induction of MMP-9 mRNA expression was found in the MC3T3-E1 cells treated with 10 mg/L Pe LPS for 24 h. The expression of MMP-9 mRNA in the 20 mg/L group was about 7 times than that in the blank control group. After 24 h, the expression of MMP-9 mRNA decreased. Maximal expression of MMP-9 protein was found in the MC3T3-E1 cells treated with 10 mg/L Pe LPS for 48 h ([35 055±2 346] ng/L) showing the highest enzyme activity. The mRNA of MMP-9 decreased significantly after pretreatment with 10 µmol/L BAY 11-7082 for 1 h.
Conclusions:Pe LPS might induce the expression of MMP-9 in MC3T3-E1 cells through the signaling of NF-κB.
