摘要
目的观察吡格列酮和胰岛素对大鼠骨髓内皮祖细胞(EPCs)的增殖能力、凋亡及分泌NO能力的影响,并探讨其机制。方法取正常SD大鼠32只,随机分为吡格列酮组和非吡格列酮组,各16只,分别给予吡格列酮和生理盐水灌胃预处理。喂养10天后,断颈处死,密度梯度离心法取骨髓单个核细胞,在M199培养液中培养扩增EPCs并进行鉴定。贴壁细胞培养4天后,将吡格列酮组EPCs消化后进一步分为两组,分别给予胰岛素(1nmol/L)或空白干预,非吡格列酮组EPCs处理同吡格列酮组EPCs。24h后检测NO水平,3天后检测凋亡情况,7天后检测细胞增殖能力。结果吡格列酮预处理能提高EPCs数量(P<0.01),吡格列酮预处理和(或)体外给予胰岛素干预组EPCs与未处理组细胞相比EPCs增殖能力提高(P<0.01),凋亡程度降低(P<0.05),培养上清中NO浓度增加(P<0.05)。结论体内吡格列酮预处理与体外胰岛素干预均能促进EPCs增殖,抑制EPCs凋亡,并促进EPCs分泌NO,且两种药物联合应用具有协同作用。
Objective To investigate the effects of pioglitazone and insulin on proliferation,apoptosis and nitric oxide （NO） secretion of endothelial progenitor cells （EPCs） derived from bone marrow. Methods Thirty-two SD rats were randomly divided into pioglitazone group,which was given pioglitazone （20 mg·kg^-1 ·d^-1） by intragastric administration for 10 d,and non- pioglitazone group,which was given physiologic saline（0. 9%）of the same volume. Each group included 16 rats. Mononuclear cells were collected from rats＇ bone marrow by density gradient centrifugation, cultured with M 199 medium,and identified by UEA-I and ac-LDL staining. Of them the double positive ones were EPCs. Once harvested, EPCs of both groups were given vehicle or insulin （1 nmol/L） respectively. After 24 h of incubation, NO secretion of EPCs was measured by the Griess assay with spectrophotometer. After incubation for 3 d,apoptosis was detected by FCM. After 7 d of incubation,proliferation capacity was measured by MTT assay. Results Pioglitazone given in vivo could increase the number of EPCs. Compared with the control group, pioglitazone and insulin group,insulin group and pioglitazone group respectively showed higher proliferation capacity（ P 〈0.01 ）, lower EPCs apoptosis （ P 〈0.05）, and more NO production （ P 〈0.05）. Conclusion Both pioglitazone given in vivo and insulin given in vitro can stimulate EPCs proliferation,decrease its apoptosis,and promote its secretion of NO,and these two kinds of intervention have synergistic effects.
出处
《中华老年心脑血管病杂志》
CAS
北大核心
2008年第3期202-205,共4页
Chinese Journal of Geriatric Heart,Brain and Vessel Diseases
关键词
胰岛素
血管内皮生长因子类
一氧化氮
细胞增殖
细胞凋亡
吡格列酮
内皮祖细胞
insulin
vascular endothelial growth factors
nitric oxide
cell proliferation
apoptosis
pioglitazone
endothelial progenitor cells
