Effect of dexmedetomidine on neuronal differentiation of umbilical cord blood mesenchymal stem cells

Abstract

Abstract:

BACKGROUND: Increasing evidence has shown that dexmedetomidine has a significant neuroprotection against ischemia-reperfusion injury, but its mechanism of action on the nervous system and its pathways to affect nerve cell function are rarely reported.
OBJECTIVE: To study the effect of dexmedetomidine on proliferation and neuronal differentiation of umbilical cord blood mesenchymal stem cells.
METHODS: Umbilical cord blood mesenchymal stem cells at passage 3 were divided into control, low-concentration (1 μg/L) and high-concentration (10 μg/L) dexmedetomidine groups. MTT was used to detect the viability of umbilical cord blood mesenchymal stem cells; western blot assay was used to determine the ERK1/2 phosphorylation in the cells; immunofluorescence staining was used to detect the percentage of NeuN/DAPI, GFAP/DAPI and Nestin/DAPI positive cells. Cell differentiation and proliferation abilities were observed.
RESULTS AND CONCLUSION: Cell viability and EKR1/2 phosphorylation level were significantly increased in the low-concentration group compared to the control group (P < 0.05); while these two indicators in the high-concentration group were significantly lower than those in the other two groups (P < 0.05). The number of NeuN and GFAP positive cells was increased significantly in the two dexmedetomidine groups compared with the control group (P < 0.05), while the number of Nestin positive cells was significantly decreased in the two
dexmedetomidine groups (P < 0.05). These findings indicate that low-concentration dexmedetomidine can induce the proliferation of umbilical cord blood mesenchymal stem cells that is however inhibited by the high-concentration dexmedetomidine, which can be realized by regulating EKR1/2 phosphorylation level. In addition, low- and high-concentration dexmedetomidine can both induce the neuronal differentiation of umbilical cord blood mesenchymal stem cells.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: fetal blood, mesenchymal stem cells, dexmedetomidine, cell proliferation

CLC Number:

R394.2

Fan Zhi-gang, Qiao Lei. Effect of dexmedetomidine on neuronal differentiation of umbilical cord blood mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(50): 8088-8092.

Figures/Tables 4

2.2 流式细胞仪检测结果 CD34、CD45的阳性率不足10%，CD19和CD86阳性率少于10%，CD90、CD105阳性率高于70%，主要组织相容性复合体HLA-ABC的阳性率为90%以上，HLA-DR阳性率低于5%，结果显示所培养细胞符合间充质干细胞表面分子标记特征。 2.3 右美托咪定对脐血间充质干细胞活力的影响 MTT法检测结果表明，低浓度右美托咪定组吸光值较对照组显著增高(P < 0.05)，高浓度右美托咪定组吸光值则较对照组明显降低(P < 0.05)，高浓度右美托咪定组吸光值较低浓度右美托咪定组也明显降低(P < 0.05)，说明低浓度右美托咪定可以促进干细胞增殖，而高浓度右美托咪定则抑制其增殖，见图2。 2.4 右美托咪定对脐血间充质干细胞增殖期ERK1/2磷酸化的影响与对照组相比，低浓度右美托咪定组脐血间充质干细胞ERK1/2磷酸化水平明显增加(P < 0.05)，而高浓度右美托咪定组脐血间充质干细胞ERK1/2磷酸化水平则较明显下降(P < 0.05)，高浓度右美托咪定组较低浓度右美托咪定组也明显下降(P < 0.05)，说明低浓度右美托咪定促进干细胞增殖可能是通过提高ERK1/2磷酸化水平实现的，而高浓度右美托咪定抑制其增殖是降低了ERK1/2磷酸化水平，见图3。2.5 右美托咪定对脐血间充质干细胞向神经细胞分化的影响通过免疫荧光计数可见，对于NeuN阳性(神经元细胞)、GFAP阳性(胶质细胞)比例，低浓度右美托咪定组和高浓度右美托咪定组均较对照组明显增加(P < 0.05)，见图4A，B；低、高浓度右美托咪定组间差异无显著性意义(P > 0.05)。对于Nestin阳性(神经干细胞)比例，低浓度右美托咪定组和高浓度右美托咪定组均较对照组明显降低(P < 0.05)，见图4C，D；低、高浓度右美托咪定组间差异无显著性意义(P > 0.05)，说明高、低浓度右美托咪定都可促进脐血干细胞向神经细胞的分化，且差异无显著性意义，见图4E。"

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