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目的:观察青心酮对内皮祖细胞的影响。方法:采用密度梯度离心法从脐血中获取单个核细胞,将其接种在人纤维连接蛋白包被培养板,培养7天后,收集贴壁细胞,加入不同浓度青心酮(分别为10-4、10-5、10-6mol/L)培养一定的时间(6、12、24、48,72 h)。通过激光共聚焦显微镜鉴定FITC-UEA-I和DiI-acLDL双染色阳性细胞被认为是正在分化的EPCs。然后分别采用MTT比色法、改良的Boyden小室、黏附能力测定实验观察EPCs的增殖能力、迁移能力及黏附能力。结果:青心酮显著增加EPCs集落形成数,并且EPCs集落形成数随青心酮浓度增加及作用时间延长而增加,10-4mol/L浓度青心酮作用72h对EPCs数量的影响最为显著。青心酮也显著改善了脐血EPCs的黏附能力、迁移能力和增殖能力。结论:结果提示青心酮可增加EPCs的数量且伴随着EPCs功能的改善。
Objective: To observe the effect of Qingxin Ketone on endothelial progenitor cells. METHODS: Mononuclear cells were obtained from umbilical cord blood by density gradient centrifugation and inoculated on human fibronectin-coated plates. After 7 days of culture, adherent cells were collected and various concentrations of carbomycin were added (10-4, respectively). , 10-5, 10-6 mol/L) were cultured for a certain period of time (6, 12, 24, 48, 72 h). Identification of FITC-UEA-I and DiI-acLDL double-stained positive cells by laser confocal microscopy was considered to be differentiation of EPCs. Then the MTT colorimetric method, the modified Boyden chamber and the adhesive ability test were used to observe the proliferation, migration and adhesion of EPCs. Results: Qingxin Ketone significantly increased the number of EPCs colony formation, and the number of colony forming EPCs increased with the increase of the concentration of Qingxin Ketone and the prolongation of action time. The effect of 10-4 mol/L Qingxin Ketone for 72 h on the number of EPCs was the most significant. Qingxin ketone also significantly improved the adhesion, migration, and proliferation of umbilical cord blood EPCs. Conclusions: The results suggest that diacyl ketone can increase the number of EPCs and is accompanied by an improvement in the function of EPCs.