Hyperglycemia metabolic memory effect in late endothelial progenitor cells from human peripheral blood

Abstract:

Objective：To explore whether there is a hyperglycemia metabolic memory in late endothelial progenitor cells（EPCs） from human peripheral blood. Methods：Peripheral blood mononuclear cells（PBMC） were separated by density gradient centrifugation. Then PBMC was induced and cultivated for two weeks to late EPCs. Immunochemical staining was used to identify cultured cells. MTT was used to explore the gradient of concentration and time of high glucose. Cells were divided into three groups，namely：normal glucose group（NG，5.5 mmol/L treatment for 6 d），high glucose group（HG，30 mmol/L treatment for 6 d），hyperglycemia memory group（MG，30 mmol/L treatment for 6 d then 5.5 mmol/L treatment for 6 d）. Adhesion test，MTT，EdU experiments，Transwell chamber，angiopoiesis experiments were used to measure the ability of adhesion，proliferation，migration，tube formation respectively. Results：The adhesion results showed that the adhesion ability was lower in high glucose（38.296±6.615） group and hyperglycemia memory group（37.815±6.196） than in normal glucose group（63.370±10.363）（P<0.05）. MTT analysis showed that the proliferation ability was lower in high glucose（79.041±19.678） group and hyperglycemia memory group（81.524±6.710） than in normal glucose group（100.000±0.000）（P<0.05）. EdU analysis showed that the proliferation ability was lower in high glucose（45.809±7.945） group and hyperglycemia memory group（37.740±8.206） than in normal glucose group（88.858±5.727）（P<0.05）. The migration results showed that the migration ability was lower in high glucose（25.111±6.051） group and hyperglycemia memory group（25.778±6.037） than in normal glucose group（64.889±10.729）（P<0.05）. The tube formation results showed that the tube formation ability was lower in high glucose（12.690±3.616） group and hyperglycemia memory group（14.198±4.073） than in normal glucose group（64.407±6.358）（P<0.05）. Compared with those of high glucose group，the adhesion，proliferation，migration，tube formation of hyperglycemia memory group was still not improved（P>0.05）. Conclusion：Late EPCs exist hyperglycemia metabolic memory.