The expression profile of microRNA in Porphyromonas gingivalis lipopolysaccharide induced inflammation in human dental pulp cells

doi:10.3877/cma.j.issn.1674-1366.2017.06.003

Abstract

Abstract:

Objective

To investigate the microRNA (miRNA) expression profile in the inflammation of human dental pulp cells (hDPCs) induced by Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) .

Methods

After isolated and cultured in vitro, the hDPCs were stimulated with Pg-LPS (10 μg/ml) for 0, 3, 6, 12, 24 h. The expression of inflammatory cytokines (IL-6, IL-8) were evaluated by reverse-transcription polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA) . Meanwhile Illumina HiSeq^TM 2500 sequencing technology was used for investigating differentially expressed miRNAs. Bioinformatic analysis was applied for predicting the target genes of miRNAs and for Gene Ontology analysis, Kyoto Encyclopedia of Genes and Genomes analysis. Data were analyzed using t test or One-Way ANOVA.

Results

The mRNA expression of IL-6 and IL-8 were increased by (7.4 ± 1.1) , (61.6 ± 20.8) times respectively at 3 h with the peak by Pg-LPS (t_IL-6=-9.268, P_IL-6= 0.001; t_IL-8=-5.047, P_IL-8= 0.037) . The protein expression of IL-6 and IL-8 were upregulated by Pg-LPS in a time dependent manner with the concentration of (44 ± 19) , (4016 ± 670) pg/ml at 24 h (t_IL-6=-3.621, P_IL-6= 0.022; t_IL-8=-4.902, P_IL-8= 0.008) . A total of 680 miRNAs were detected with expression changed after stimulated with Pg-LPS (10 μg/ml) , and 2 down-regulated miRNAs and 18 up-regulated miRNAs were screening as differentially expressed miRNAs. Gene function enrichment analysis showed that differentially expressed miRNAs were involed in inflammatory related pathway such as Wnt signal pathway, TGF-β signal pathway, PI3K-Akt signal pathway, MAPK signal pathway.

Conclusion

Our data identified differential expression of miRNAs after hDPCs treated with Pg-LPS, indicating miRNAs may involed in the inflammation in hDPCs induced by Pg-LPS.

Key words: Porphyromonas gingivalis, Lipopolysaccharides, Dental pulp, Inflammatory cytokines, MicroRNA

Zehuan Mo, Qiong Xu. The expression profile of microRNA in Porphyromonas gingivalis lipopolysaccharide induced inflammation in human dental pulp cells[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2017, 11(06): 333-340.

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References 42

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基因	引物序列
IL-8	F:5′-GGTGCAGTTTTGCCAAGGAG-3′
?	R:5′-TTCCTTGGGGTCCAGACAGA-3′
IL-6	F:5′-TGCAATAACCACCCCTGACC-3′
?	R:5′-AGCTGCGCAGAATGAGATGA-3′
GAPDH	F:5′-TCTCCTCTGACTTCAACAGCGACA-3′
?	R:5′-CCCTGTTGCTGTAGCCAAATTCGT-3′

基因	引物序列
IL-8	F:5′-GGTGCAGTTTTGCCAAGGAG-3′
?	R:5′-TTCCTTGGGGTCCAGACAGA-3′
IL-6	F:5′-TGCAATAACCACCCCTGACC-3′
?	R:5′-AGCTGCGCAGAATGAGATGA-3′
GAPDH	F:5′-TCTCCTCTGACTTCAACAGCGACA-3′
?	R:5′-CCCTGTTGCTGTAGCCAAATTCGT-3′

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