高糖对成人外周血平滑肌祖细胞功能的影响  

Effects of high glucose on biological characteristics of smooth muscle progenitor cells from human peripheral blood

作　　者：周学凯[1] 倪旭东[1] 李飞[1] 张荣庆[1] 郭文怡[1] 

机构地区：[1]第四军医大学西京医院心血管内科,西安710032

出　　处：《岭南心血管病杂志》2011年第4期311-315,共5页South China Journal of Cardiovascular Diseases

基　　金：陕西省科学技术研究发展计划项目(项目编号:2010K01-171课题名称:冠脉介入治疗术后再狭窄防治的基础与临床研究)

摘　　要：目的观察不同浓度葡萄糖对体外培养成人外周血平滑肌祖细胞(smooth muscle progenitor cells,SPCs)增殖、迁移和黏附能力的影响。方法采用密度梯度离心法从健康成人外周血获取单个核细胞,培养12 d后,采用流式细胞仪对诱导扩增的SPCs进行分析鉴定并分选纯化。间接免疫荧光染色法观察SPCs培养到第28天后人平滑肌细胞特异性肌动蛋白(α-SMA)的表达情况。收集培养第12天的SPCs,随机(补充具体的随机方法?)分为5组并给予不同浓度葡萄糖干预:对照组(5.5 mmol/L),11 mmol/L组,22 mmol/L组,44 mmol/L组和渗透压对照组(5.5 mmol/L葡萄糖加38 mmol/L甘露醇)。分别用噻唑蓝(methyl thiazolyl tetrazolium,MTT)比色法,Transwell小室迁移实验以及黏附能力测定实验检测各组干预6 d后SPCs增殖、迁移和黏附能力的变化。此外,培养SPCs 8 d,期间用22 mmol/L葡萄糖分别干预0、2、4、8 d,观察各组细胞增殖、迁移和黏附能力的变化。结果与对照组相比,高糖各组均能明显促进外周血SPCs的增殖、迁移和黏附能力,其中22 mmol/L葡萄糖组的影响最为显著,44 mmol/L葡萄糖组的促进作用有所下降。用22 mmol/L葡萄糖分别干预SPCs 0、2、4、8 d,其增殖、迁移和黏附能力随着作用时间延长而增强,以干预8 d组最显著。结论高糖能在一定范围内增强外周血SPCs的增殖、迁移、黏附能力,随着浓度增加和时间延长,作用更明显。推测长期高血糖通过促进SPCs的功能参与受损血管过度修复,引起部分心血管疾病的发生发展。Objectives To investigate the effects of high glucose on proliferation, migration and adhesion of smooth muscle progenitor cells （SPCs） in vitro. Methods Mononuclear cells were isolated from adult collected peripheral blood by density gradient centrifugation and cultured. After 12 days of culture in vitro, SPCs were identified and purified as adherent ceils double positive for CD14 and CD105, then sorted by flow cytometry. Twenty-eight days later, immunofluorescence was used to analyze the expression of ct-smooth muscle actin（ot-SMA）. The SPCs were harvested at the 12th day and randomly divided into 5 groups （control group, 11 mmol/L group, 22 mmol/L group, 44 mmol/L group and osmotic pressure control group）. The SPCs were incubated with glucose in a series of concentrations for 6 days. Proliferation and migration of SPCs were assayed by methyl thiazolyl tetrazolium （MTr） assay and Transwell chamber assay respectively. SPCs adhesion assay was performed by replating the cells on fibronectin-coated dishes and the adherent cells were then counted. In addition, the 8-day SPCs were incubated with glucose （22 mmol/L） for different durations （0,2,4,8 days）. And its proliferation and migration ability were abserved and adhesion assay was performed. Results Compared with control group, high concentration of glucose accelerated the proliferation, migration and adhesion ability of SPCs, especially when the SPCs were incubated with glucose in the concentration of 22 mmol/L for 8 days. Conclusions To some extent, high concentration of glucose could significantly improve the function of SPCs in a concentration and time dependent manner. It could be speculated that long-term hyperglycemia in patients could stimulate the function of SPCs, which plays an important role in the development of cardiovascular disease.

关 键 词：平滑肌祖细胞 葡萄糖 增殖 迁移 黏附 

分 类 号：R587.1[医药卫生—内分泌]