摘要
目的观察过氧化物酶增殖激活受体γ（peroxisome proliferator-activated receptor-γ,PPARγ）激活后对内皮祖细胞（endothelial progenitor cells,EPCs）侵袭能力的影响并探讨其分子机制。方法分离和培养大鼠骨髓源性EPCs,分别用PPARγ激动剂罗格列酮（0、1、5、10μmol/L）、PPARγ拮抗剂GW9662（5μmol/L）或组织蛋白酶L（cathepsin L）抑制剂Z-FF-FMK（10μmol/L）干预EPCs后,采用基质胶侵袭实验检测EPCs的侵袭能力;半定量RT-PCR、Western blot检测cathepsin L mRNA和蛋白的表达。结果 PPARγ激动剂罗格列酮（1~10μmol/L）呈剂量依赖增强EPCs的侵袭能力、上调EPCs中cathepsin L的mRNA和蛋白表达（P〈0.05,P〈0.01）,而PPARγ拮抗剂GW9662（5μmol/L）可削弱罗格列酮（10μmol/L）上调cathepsin L mRNA和蛋白表达的作用（P〈0.01）;此外,GW9662（5μmol/L）或cathepsin L抑制剂Z-FFFMK（10μmol/L）均可明显减弱罗格列酮（10μmol/L）促进EPCs侵袭的作用（P〈0.01）。结论 PPARγ激活后通过上调cathepsin L表达增强EPCs的侵袭能力。
Objective To observe the effect of peroxisome proliferator-activated receptor-γ （PPARγ） activation on the invasive capacity of endothelial progenitor cells （EPCs） and explore the underlying mechanism. Methods The rat bone marrow-derived EPCs isolated and cultured in vitro were treated with PPARγ agonist rosiglitazone （0, 1, 5, and 10 μmol/L）, PPARγ antagonist GW9662 （5 μmol/L） or cathepsin L inhibitor Z-FF-FMK （10 μmol/L）. The invasive capacity of EPCs was evaluated by Matrigel invasion assay. The mRNA and protein expression of cathepsin L were analyzed by semi-quantitative RT-PCR and Western blotting. Results Treatment of PPARγ agonist rosiglitazone increased the invasive capacity of EPCs in a dose-dependent manner （P〈0.05, P〈0.01）. The mRNA and protein expression of cathepsin L were up-regulated by rosiglitazone in a concentration-dependent manner （P〈0.05, P〈0.01）, which was blocked by PPARγ antagonist GW9662 （5 μmol/L） （P〈0.01）. In addition, pretreatment of EPCs with GW9662 （5 μmol/L） or cathepsin L inhibitor Z-FF-FMK （10 μmol/L） attenuated rosiglitazone-promoted EPCs invasion （P〈0.01）. Conclusion These findings demonstrate that the activation of PPARγ may enhance the invasive capacity of EPCs via up-regulating cathepsin L expression.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2014年第16期1643-1646,共4页
Journal of Third Military Medical University
基金
国家自然科学基金(81100112)~~
关键词
过氧化物酶增殖激活受体γ
内皮祖细胞
侵袭
归巢
组织蛋白酶L
peroxisome proliferator-activated receptor-γ
endothelial progenitor cells
invasion
homing
cathepsin L
