Effect of anti-tumor necrosis factor antibody on TRAIL and TNF-α expression induced by lipopolysaccharide in human periodontal ligament fibroblasts
CHEN Yan, YE Bin, MOU Wen-yuan, YU Cheng-bo, LIN Bu, YIN Xiang-peng, XU Hong-miao
2015, 20(5):
493-498.
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AIM: To investigate the possible roles of tumor necrosis factor(TNF) superfamily in the pathogenesis of periodontal disease inflammation, the effect of anti-tumor necrosis factor antibody on TRAIL and TNF-α expression induced by lipopolysaccharide in human periodontal ligament fibroblasts is observed.METHODS: HPLFs were cultured to the sixth generation cells and were interfered with defferent concentrations of lipopolysaccharide(0,0.1,1,10,100 μg/mL,respectively) for 24 h, named group Z0,Z0.1,Z1,Z10 and Z100. HPLFs were also cultured with 1 μg/mL lipopolysaccharide and 75 μg/mL anti- TNF antibody (named group Z1+75). The expressions of both TNF-α and TRAIL mRNA were analysed by Real-time quantitative reverse transcription polymerase chain reaction(QRT-PCR),while TRAIL protein were determined by West Blot methods.RESULTS: The expressions of TRAIL mRNA in group Z1 or Z10 were significantly higher than those in group Z0 or Z0.1 (all P<0.05),in group Z100 were significantly higher than those in group Z0(P<0.05).But there were no significant differences neither between group Z1 and Z10 nor among group Z100, Z0.1, Z1 and Z10 (all P>0.05). Moreover, the expressions of TNF-α mRNA in group Z1 or Z10 were significantly higher than those in group Z0 or Z100 (all P<0.05), whereas there were nonsignificant differences neither between group Z1 and Z10 nor among group Z0, Z0.1, and Z100 (all P>0.05). Dramatically, the expressions of TRAIL protein were significant difference between each groups, in group Z100 <Z0 < Z0.1 <Z1(all P<0.05 or P<0.01).And it was significantly higher in group Z10 than those in group Z0 or Z100(all P<0.01),while there were nonsignificant differences neither between group Z10 and Z0.1 nor among group Z10 and Z1 (all P>0.05). Noteworthy, the expressions of either TRAIL mRNA and protein or TNF-α mRNA in group Z1+75 were significantly decreased than those in group Z1(P<0.05 or <0.01,respectively). Furthermore,there were significance positive correlation between levels of TRAIL and TNF-α mRNA(r=0.819,n=30, P<0.01).CONCLUSION: The results shows that the expressions of TRAIL and TNF-α in HPLFs can be induced by lipopolysaccharide, the levels of them were elevated in the first and decreased subsequently, this changes were accompanied by lipopolysaccharide concentration. Date also show the function of lipopolysaccharide can be inhibited by anti-TNF antibody.