Hypoxia induces the migration of endothelial progenitor cells

doi:10.3969/j.issn.2095-4344.0810

Abstract

Abstract:

BACKGROUND: Local endothelial progenitor cells (EPCs) have been reported to promote migration and homing of EPCs by releasing stromal cell-derived factor-1 (SDF-1) under hypoxia environment.
OBJECTIVE: To investigate the effect of different hypoxia periods on the expression of SDF-1 protein in EPCs and the migration of EPCs, thus providing new ideas for the treatment of ischemic diseases.
METHODS: The bone marrow of the long bone was removed from Sprague-Dawley rats used for isolate EPCs. After culture and identification, EPCs were divided into four groups: control (normoxia), 6-, 12- and 24-hour hypoxia groups. The expression level of SDF-1 in the supernatant of EPCs in each group was detected by ELISA and immunofluorescence. The mRNA and protein levels of SDF-1 were detected by RT-PCR and western blot assay. The effect of hypoxia on the migration of EPCs was tested by Transwell assay.
RESULTS AND CONCLUSION: The obtained cells were identified as EPCs. ELISA and immunofluorescence results showed that compared with the control group, the expression level of SDF-1 began to increase at 6 hours of hypoxia, peaked at 12 hours of hypoxia, and then decreased at 24 hours of hypoxia. RT-PCR and western blot assay results found that the mRNA and protein expression levels of SDF-1 were the highest in the 12-hour hypoxia group, followed by 6- and 24-hour hypoxia groups, and the lowest in the control group (P < 0.01). The number of EPCs penetrating the membrane in the 12-hour hypoxia group was significantly more than that in the 6- and 24-hour hypoxia groups, and the number in the 24-hour hypoxia group was significantly more than that in the control group (P < 0.05). In summary, hypoxia can promote the secretion of SDF-1 and induce the migration of EPCs.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Cell Hypoxia, Endothelial Cells, Cell Movement, Tissue Engineering

CLC Number:

R318

Zhao Hui, Gao Yu-zhong. Hypoxia induces the migration of endothelial progenitor cells[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(12): 1902-1908.

Figures/Tables 2

2.3 ELISA检测不同条件下基质细胞衍生因子1的表达低氧 6，12，24 h诱导基质细胞衍生因子1的表达，6 h表达开始增强，12 h表达最强，24 h又有所减弱。与常氧对照组比较，低氧组基质细胞衍生因子1的表达显著增高(P < 0.05)；低氧12 h显著高于低氧6 h和24 h(P < 0.05)。见表2。 2.4 免疫荧光检测不同条件下基质细胞衍生因子1的表达低氧6，12，24 h诱导基质细胞衍生因子1的表达，与常氧对照组比较，低氧6 h表达开始增强，12 h表达最强，24 h又有所减弱。因基质细胞衍生因子1为胞质和胞膜表达，FITC为绿色荧光。见表3，图3。 2.5 RT-PCR检测不同条件下基质细胞衍生因子1 mRNA的表达分析RT-PCR条带图可知，低氧6，12，24 h诱导基质细胞衍生因子1的表达，与常氧对照组比较，低氧6 h表达开始增强，12 h表达最强，24 h又有所减弱，见图4。分析表4可知，与常氧对照组相比，低氧6，12，24 h诱导基质细胞衍生因子1 mRNA的表达显著上调(P < 0.01)。 2.6 Western blot检测不同条件下基质细胞衍生因子1蛋白的表达分析图5 Western blot条带和可知，与常氧对照组相比，低氧6，12，24 h诱导条件下基质细胞衍生因子1蛋白的表达增加，其中，低氧12 h条件下基质细胞衍生因子1蛋白的表达最强，依次为低氧24 h和低氧6 h，对基质细胞衍生因子1和β-actin蛋白积分光密度进行计算，将两者比值作为基质细胞衍生因子1的相对表达量，结果与条带的表达情况相符合，说明Western blot检测结果与ELISA检测和免疫荧光检测结果相吻合，见表5。 2.7 Transwell检测低氧对内皮祖细胞迁移的影响分析表6可知，与常氧对照组比较，低氧组24 h穿膜细胞数显著增加(P < 0.05)；低氧12 h组显著多于低氧6 h组和低氧24 h组，差异有显著性意义(P < 0.05)。见图6。"

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