Effect of hypoxia on the expression of matrix metalloproteinase and tissue inhibitors of matrix metalloproteinase mRNA in human periodontal ligament fibroblasts in vitro

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Author:
SONG Ai-mei(Department of Periodontology, School of Stomatology, Shandong University; Shandong Provincial Key Laboratory of Oral Biomedicine, Jinan 250012,China)
HOU Chao()
CHEN Jia-fang(Department of Periodontology, School of Stomatology, Shandong University; Shandong Provincial Key Laboratory of Oral Biomedicine, Jinan 250012,China)
SUN Jing(Department of Periodontology, School of Stomatology, Shandong University; Shandong Provincial Key Laboratory of Oral Biomedicine, Jinan 250012,China)
TIAN Tian(Department of Periodontology, School of Stomatology, Shandong University; Shandong Provincial Key Laboratory of Oral Biomedicine, Jinan 250012,China)
LI Shu(Department of Periodontology, School of Stomatology, Shandong University; Shandong Provincial Key Laboratory of Oral Biomedicine, Jinan 250012,China)
Journal Title:
Chinese Journal of Stomatology
Issue:
Volume 47, Issue 10, 2012
DOI:
10.3760/cma.j.issn.1002-0098.2012.10.006
Key Word:
Periodontal ligament ; Fibroblasts ; Anoxia; Matrix metalloproteinases ; Tissue inhibitors of matrix metalloproteinases

Abstract: Objective To investigate the effect of hypoxia on the expression of matrix metalloproteinase(MMP) and tissue inhibitors of matrix metalloproteinase (TIMP) in human periodontal ligament fibroblasts(HPDLF).Methods HPDLF were cultured in α-minima essential medium(α-MEM) and subcultured at confluence. In the hypoxic groups,cells were incubated in a humidified atmosphere of 1%O2,5%CO2,94% N2 at 37 ℃ for 12,24 and 48 h,respectively.In the normoxic control group,cells were incubated under normoxic conditions of 20% O2,5% CO2,75% N2.The mRNA expression of MMP and TIMP was measured using reverse transcription-polymerase chain reaction (RT-PCR).The data was analyzed by Student's t test,one-way ANOVA and LSD test with SPSS 13.0 software package. Results The expression of MMP-2,TIMP-1 and TIMP-2 mRNA in the hypoxia groups was higher than that in control.The expression of MMP-2 mRNA in hypoxic groups showed a significantly increasing trend.There was significant difference between the hypoxic group and the normoxic control group in the expression of MMP-2 mRNA in HPDLF(P < 0.01 ).The expression of TIMP-1,TIMP-2 mRNA in hypoxic groups of 12 h was momentarily increased. There was significant difference between the hypoxic 12 h group and the normoxic control group in the expression of TIMP-1,TIMP-2 mRNA in HPDLF( P <0.05 ).However,with prolonged hypoxia time,the expression of TIMP-1,TIMP-2 mRNA in hypoxic groups showed a significantly declining trend,there were significant differences between the hypoxic 12,24 and 48 h group and the normoxic control group in the expression of TIMP-2 mRNA in HPDLF( P < 0.05 ).The expression of MMP-1 mRNA in hypoxic groups of 12 h was momentarily decreased and then increased after 24 h of hypoxia.There were significant differences between the hypoxic 48 h group and the normoxic control group in the expression of MMP-1 mRNA in HPDLF(P <0.05).There were significant differences between the hypoxic 12 h group and the normoxic control group in the ratio of MMP-1/TIMP-1 mRNA ( P < 0.05 ).The ratio of MMP-2/ TIMP-2 mRNA in the hypoxia group significantly increased compared with normoxic group. There were significant differences between the hypoxic group and the normoxic control group in the ratio of MMP-2/TIMP-2 mRNA(P < 0.05).Conclusions Hypoxia could change the expression of MMP and TIMP mRNA and other relevant growth factors and also lead to the imbalance of MMP-2/TIMP-2 mRNA expression.It is suggested that the imbalance of MMP-2/TIMP-2 expression may be closely correlated with the occurrence and development of periodontal disease and play an important role in the process of periodontal tissue destruction in periodontitis.

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