原发性高血压条件下骨髓来源内皮祖细胞数量及功能的变化

Number and function of bone marrow-derived endothelial progenitor cells in essential hypertension

目的了解原发性高血压对体外培养骨髓来源内皮祖细胞(EPC)数量及功能的影响。方法将自发性高血压大鼠(SHR)分为6、12周龄组,以同周龄正常血压Wistar-Kyoto(WKY)大鼠为对照。以密度梯度离心法分离骨髓来源单个核细胞体外培养,差速贴壁法培养7 d后收集贴壁细胞并行鉴定为骨髓来源内皮祖细胞,测定EPC数量、细胞增殖活力(MTT法)、NO分泌及细胞凋亡率。结果 6周龄SHR骨髓来源EPC数量与6周龄WKY大鼠比较差异无统计学意义(P>0.05),细胞增殖能力、NO分泌能力受损(P<0.05),凋亡率稍增多但差异无统计学意义(P>0.05)。12周龄SHR骨髓来源EPC数量与12周龄WKY大鼠比较明显减少(P<0.01),细胞增殖能力、NO分泌能力受损(P<0.01),凋亡增多(P<0.05)。结论随着SHR血压升高和原发性高血压病程延长,骨髓来源内皮祖细胞数量减少,功能减退,凋亡增多。

Objective To evaluate the effect on number and function of endothelial progenitor cells in essential hypertension condition. Methods Mononuclear cells of 6-week and 12-week spontaneously hypertensive rats （SHR） from rat bone marrows were isolated by density gradient centrifugation, and age matched Wistar-Kyoto rats （WKY） were used as controls. After 7 d cultivation, adhering cell, and identified the cell to be EPC were harvested by FITC-UEA-1 and Dil-ac-LDL double staining. Number of EPC were counted, and detected proliferative capacity, NO secretion and apoptosis rate of EPC. Results There was no significant difference （P 〉 0.05） of bone marrow-derived EPC number between 6-week SHR and 6-week WKY, the capacity of proliferation and NO secretion were impaired （P 〈 0.05）, rate of cell apoptosis was unremarkablely higher than that of controls （P 〉 0.05）. While EPC number of 12-week SHR was much less than 12-week WKY. The capacity of proliferation and NO secretion were significantly impaired （P 〈 0.01）, rate of cell apoptosis was higher than control （P 〈 0.05）. Conclusion In accordance with the development of high blood pressure and hypertension progress, number of bone marrow-derived EPC become less and the function of EPC are impaired.