促红细胞生成素对人牙髓细胞迁移能力的作用

目的研究促红细胞生成素（EPO）对人牙髓细胞（hDPC）迁移能力的影响，并初步探讨相关分子机制。 方法实时荧光定量聚合链反应（PCR）检测EPO对hDPC表达趋化因子mRNA的影响；Transwell实验观察不同浓度的EPO对hDPC迁移能力的影响；Western blot检测不同时间点hDPC中p38、ERK1/2、JNK磷酸化水平的变化；细胞划痕实验观察丝裂原活化蛋白激酶（MAPK）信号通路抑制剂对EPO诱导hDPC迁移的影响。 结果EPO上调趋化因子CXCR4、SDF-1 mRNA的表达（t_CXCR4= 5.727，P_CXCR4= 0.005；t_SDF-1= 3.412，P_SDF-1= 0.027）；与对照组相比，EPO显著促进hDPC的迁移能力（F= 207.775，P_{10 U/ml}= 0.000，P_{20 U/ml}= 0.000，P_{40 U/ml}= 0.000）；EPO可升高MAPK信号通路中关键蛋白ERK1/2（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）和p38的磷酸化程度（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）；MAPK信号通路抑制剂U0126、SB203580可抑制EPO诱导的hDPC迁移（t_EPO-U0126= 2.422，P_{EPO- U0126}= 0.025；t_EPO-SB203580= 3.837，P_EPO-SB203580= 0.001）。 结论EPO上调趋化因子CXCR4和SDF-1 mRNA的表达，通过激活MAPK信号通路，促进hDPC迁移。

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

ObjectiveTo investigate the effect of erythropoietin on migration of human dental pulp cells and preliminarily explore the mechanisms. MethodsThe 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. ResultsThe 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) . ConclusionEPO upregulated mRNA expression of chemotaxis factors CXCR4 and SDF-1, and enhanced hDPCs migration through activating MAPK pathway.