自发性高血压大鼠骨髓来源内皮祖细胞数量、功能及吡格列酮的预处理

Effect of pioglitazone preconditioning on quantity and function of bone marrow-derived endothelial progenitor cells in spontaneously hypertensive rats

背景:研究证明过氧化物酶体增殖物激活受体γ具有上调内皮祖细胞的功能。目的:验证过氧化物酶体增殖物激活受体γ激动剂吡格列酮对体外培养自发性高血压大鼠骨髓来源内皮祖细胞数量及功能的影响。设计、时间及地点:随机对照动物实验,于2007-05/08在解放军第四军医大学西京院心脏内科国家重点实验室完成。材料:选取清洁级自发性高血压大鼠及周龄、体质量相匹配的正常血压WKY大鼠各20只。方法:随机将自发性高血压大鼠及正常血压WKY大鼠各分为药物组和对照组,每组10只。分别给予吡格列酮(20mg/kg·d)或等量生理盐水预处理14d。于试验开始前1d和结束当天分别测各组大鼠尾动脉收缩压。以密度梯度离心法分离骨髓来源单个核细胞体外培养,差速贴壁法培养收集贴壁细胞。主要观察指标:鉴定内皮祖细胞;测定内皮祖细胞数量;采用四甲基偶氮唑盐法检测细胞增殖活力及一氧化氮分泌功能。结果:吡格列酮治疗后,正常WKY大鼠内皮祖细胞数量、增殖及一氧化氮分泌功能无明显变化;自发性高血压大鼠内皮祖细胞数量增加,增殖能力增强,一氧化氮分泌增加,3项与对照组比较,差异均具有显著性意义(P<0.01,P<0.05,P<0.01)。结论:吡格列酮预处理能上调其骨髓来源内皮祖细胞数量,并通过促进其增殖和一氧化氮分泌功能,对内皮祖细胞有保护作用。

BACKGROUND： Peroxisome proliferators-activated receptor gamma can up-regulate endothelial progenitor cells （EPCs）. OBJECTIVE： To investigate the effect of pioglitazone on quantity and function of hone marrow-derived EPCs of in vitro cultured rats with spontaneously hypertensive. DESIGN, TIME AND SETTING： The randomized control animal study was performed at State Key Laboratory of Department of Cardiology, Xijing Hospital, Fourth Military Medical University of Chinese PLA from May to August 2007. MATERIALS： Twenty clean spontaneously hypertensive and Wistar-Kyoto （WKY） rats were selected each, who had matched age and body mass. METHODS： Spontaneously hypertensive and WKY rats were respectively randomized into a drug group and a control group, with 10 rats in each group and received pioglitazone （20 mg/kg d） or saline for 14 days. Systolic pressure of the tail artery in each rat was measured at days 1 and 14 in both groups. Mononuclear cells were harvested from rat bone marrow by density gradient centrifugation. Adherent cells were collected by differential velocity adherent technique. MAIN OUTCOME MEASURES： EPC quantity; Proliferative activity and nitric oxide levels in EPCs were identified by MTT. RESULTS： There was no significant difference in EPC quantity, proliferation and nitric oxide secretion in WKY rats after treatment of pioglitazone. Pioglitazone significantly increased EPC quantity, proliferation and nitric oxide secretion in spontaneously hypertensive rats （P 〈 0.01, P 〈 0.05, P 〈 0.01 ）. CONCLUSION： Pioglitazone up-regulates bone marrow-derived EPC number, and protects EPCs by promoting EPC proliferation and nitric oxide secretion.