蛋白酶体抑制剂MG132对外周血内皮祖细胞数量与功能的影响

目的观察蛋白酶体抑制剂MG132对外周血内皮祖细胞(EPCs)数量与功能的影响。方法采用Ficoll密度梯度离心法从外周血获得单个核细胞,接种于包被人纤维连接蛋白的培养板,收集贴壁细胞,加入不同浓度MG132(20nmol/l,50nmol/l,100nmol/l,200nmol/l)培养12、24、48h。激光共聚焦显微镜鉴定,倒置荧光显微镜计数。分别采用MTT比色法、改良的Boyden小室、黏附能力测定观察EPCs的增殖、迁移、黏附能力。结果低剂量范围内,EPCs数量随MG132浓度与作用时间增加而减少,其增殖、迁移、黏附能力亦随MG132浓度与作用时间增加而减少。200nmol/l浓度MG132作用48h对EPC数量、增殖功能及黏附能力的影响最为显著。结论低剂量MG132减少EPCs数量并抑制EPCs的增殖、黏附、迁移能力。

Effects of Proteasome Inhibitor MG132 on number and Activity of Endothelial Progenitor cells from Peripheral Blood

Objective To investigate the effects of low-dose proteasome inhibitor MG132 on the number and function of peripheral blood endothelial progenitor cells （EPCs）. Methods Mononuelear ceils （MNCs） were isolated from peripheral blood by Fieoll density gradient eentrifugation, and then the MNCs were plated on fibronectin-eoated culture dishes. After 7days culture , adherent cells were treated with low-dose proteasome inhibitor MG132 in a series of final concentrations of 20nmol/1,50nmol/1,100 nmol/1,200nmol/1 for 12,24,48h. EPCs were identified as adherent cells double positive for DiLDL-up-take and leefin binding by direct fluorescent staining under a laser scanning confocal microscope. EPCs proliferation, adhesion and migration were assayed with MTT, adhesion assay and modified Boyden chamber assay, respectively. Results Incubation of isolated human MNCs with low-dose proteasome inhibitor MG132 decreased the number of EPCs, and MG132 also decreased EPCs proliferative, adhesive and migration capacity in a concentration-and time-dependent manner. Conclusions Low-dose proteasome inhibitor MG132 was found to decrease the numbert, proliferation, adhesive and migratory capacities of the EPCs.