The study on the role of erythropoietin in migration of human dental pulp cells

doi:10.3877/cma.j.issn.1674-1366.2016.03.003

Abstract

Abstract:

Objective

To investigate the effect of erythropoietin on migration of human dental pulp cells and preliminarily explore the mechanisms.

Methods

The mRNA levels of chemotaxis factors were measured by quantitative polymerase chain reaction (qPCR) . Transwell migration assay was conducted to estimate the effect of EPO on the migration of hDPCs. The phosphorylated levels of ERK, p38 and JNK in hDPCs after stimulation of EPO within 120min were determined by western blot. Wound-healing migration assay was conducted to observe the effect of MAPK inhibitors on migration induced by EPO in hDPCs.

Results

The mRNA expression of CXCR4 and SDF-1 were enhanced by EPO (t_CXCR4= 5.727, P_CXCR4= 0.005; t_SDF-1= 3.412, P_SDF-1= 0.027) . The migration ability of hDPCs was improved under EPO in comparison to control group (F= 207.775, P_{10 U/ml}= 0.000, P_{20 U/ml}= 0.000, P_{40 U/ml}= 0.000) . Both p-ERK (t_{15 min}= 6.554, P_{15 min}= 0.000; t_{30 min}= 17.305, P_{30 min}= 0.000; t_{60 min}= 8.913, P_{60 min}= 0.000; t_{120 min}=-5.896, P_{120 min}= 0.934) and p-p38 (t_{15 min}= 4.396, P_{15 min}= 0.004; t_{30 min}= 6.447, P_{30 min}= 0.000; t_{60 min}= 34.676, P_{60 min}= 0.000; t_{120 min}= 4.689, P_{120 min}= 0.003) were upregulated within 120 min after EPO treatment. Wound-healing migration assay showed that the migration ability of hDPCs was inhibited by U0126 and SB203580 pretreatment (t_EPO-U0126= 2.422, P_{EPO- U0126}= 0.025; t_EPO-SB203580= 3.837, P_EPO-SB203580= 0.001) .

Conclusion

EPO upregulated mRNA expression of chemotaxis factors CXCR4 and SDF-1, and enhanced hDPCs migration through activating MAPK pathway.

Key words: Erythropoietin, Dental pulp cell, Migration, MAPK pathway

Junyu Zeng, Qimei Gong, Junqi Ling. The study on the role of erythropoietin in migration of human dental pulp cells[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2016, 10(03): 166-171.

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Figures/Tables 5

References 18

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基因	引物序列
MCP?1	正向：5′?CCAAAGAAGCTGTGATCTTCAA?3′
?	反向：5′?TGGAATCCTGAACCCACTTC?3′
FGF2	正向：5′?CGTGCTATGAAGGAAGATGGA?3′
?	反向：5′?TGCCCAGTTCGTTTCAGT?3′
CXCR4	正向：5′?TACACCGAGGAAATGGGCTCA?3′
?	反向：5′?AGATGATGGAGTAGATGGTGGG?3′
SDF?1	正向：5′?GTCAGCCTGAGCTACAGATGC?3′
?	反向：5′?CACTTTAGCTTCGGGTCAATG?3′

基因	引物序列
MCP?1	正向：5′?CCAAAGAAGCTGTGATCTTCAA?3′
?	反向：5′?TGGAATCCTGAACCCACTTC?3′
FGF2	正向：5′?CGTGCTATGAAGGAAGATGGA?3′
?	反向：5′?TGCCCAGTTCGTTTCAGT?3′
CXCR4	正向：5′?TACACCGAGGAAATGGGCTCA?3′
?	反向：5′?AGATGATGGAGTAGATGGTGGG?3′
SDF?1	正向：5′?GTCAGCCTGAGCTACAGATGC?3′
?	反向：5′?CACTTTAGCTTCGGGTCAATG?3′

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