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-  2016 

苯妥英钠对大鼠骨髓间充质干细胞与血管内皮细胞VEGF和SCF分泌的影响
Effects of phenytoin on VEGF and SCF in rat bone marrow mesenchymal stem cells and vascular endothelial cells co-culture system

DOI: 10.7652/jdyxb201602012

Keywords: 大鼠,骨髓间充质干细胞,血管内皮细胞,间接共培养,血管新生,苯妥英钠
rat
,bone marrow mesenchymal stem cells (rBMSCs),vascular endothelial cells (rVECs),indirect co-culture,angiogenesis,phenytoin (PHT)

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Abstract:

摘要:目的 研究苯妥英钠(PHT)对大鼠骨髓间充质干细胞(rBMSCs)和血管内皮细胞(rVECs)间接共培养体系中血管内皮细胞生长因子(VEGF)和干细胞因子(SCF)分泌的影响。方法 分别建立rBMSCs、rVECs单独培养组及共培养组,各组添加不同质量浓度的PHT(0、20、40μg/mL)。在培养第2、4、6天时,用双抗体夹心ABC-ELISA法检测培养上清中VEGF和SCF的含量。结果 ①VEGF:培养第二天时,在相同PHT作用下,共培养组VEGF含量高于rBMSCs单独培养组(P<0.05)和rVECs单独培养组(P<0.001);共培养组随着培养时间的延长和PHT浓度增大VEGF含量增加(P<0.001,P<0.05)。②SCF:共培养组SCF含量高于相同PHT浓度下的单独培养组;在共培养组中,当PHT为20μg/mL时,随着培养时间延长SCF含量增加;当PHT为40μg/mL时,随着培养时间延长SCF含量减少,但差异无统计学意义(P>0.05)。结论 PHT可促进rBMSCs与rVECs共培养体系中VEGF的分泌,可能降低SCF的分泌。
ABSTRACT: Objective To investigate the effects of phenytoin (PHT) on the secretion of vascular endothelial growth factor (VEGF) and stem cell factor (SCF) based on the establishment of indirect co-culture system of rat bone marrow mesenchymal stem cells (BMSCs) and vascular endothelial cells (VECs). Methods Indirect co-culture model of rat BMSCs and VECs was established. Experimental groups: indirect co-culture groups (PHT concentrations were 0, 20 and 40μg/mL); the control group: BMSCs culture group and VECs culture group (PHT concentrations were 0, 20 and 40μg/mL). The contents of VEGF and SCF in the culture supernatant were measured using double antibody sandwich ABC-ELISA method on cultivation days 2, 4, 6. Results ELISA assay of the rBMSCs and rVECs in indirect co-culture supernatants, collected on culture days 2, 4 and 6 showed that: ① VEGF: On culture day 2, VEGF level in the co-culture groups was significantly higher than those in BMSCs group (P<0.05) and rVECs group (P<0.001). As culture time prolonged and PHT concentration increased to 20μg/mL and 40μg/mL, VEGF level increased too (P<0.001, P<0.05). ② SCF testing results showed that the secretion of SCF in co-culture groups was higher than that in the control groups. When PHT was 20μg/mL, the secretion of SCF increased as the incubation time increased; but as the incubation time increased, PHT concentration of 40μg/mL made SCF content decrease. Each group did not significantly differ (P>0.05). Conclusion PHT promotes the secretion of VEGF and may reduce the secretion of SCF

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