摘要
目的:检验催产素能否将去分化脂肪(DFAT)细胞诱导分化成为心肌样细胞。方法:从脂肪中分离成熟的脂肪细胞,通过天花板培养使成熟脂肪细胞去分化为DFAT细胞。按照不同诱导的时间(1周、2周、3周)及低中高3个不同浓度的(1×10-8mol/L、1×10-7mol/L、1×10-6mol/L)分组对DFAT进行诱导培养。通过免疫荧光检测法、RT-PCR及Western blot分别在基因水平和蛋白水平上检测心肌特异性标记的表达,以人类心肌组织作为阳性对照,未加诱导的DFAT细胞作为阴性对照。结果:高浓度的催产素(1×10-6mol/L)造成实验细胞大量死亡,无法进行后续试验。以1×10-8mol/L及1×10-7mol/L浓度的催产素诱导3周后,DFAT细胞在基因及蛋白水平上检测到心脏特异性标记GATA4、Nkx2.5及cTnT的表达,但未发现自主搏动现象。结论:生理浓度和中等浓度的催产素,可以将DFAT细胞诱导成为心肌样细胞。
AIM： To explore whether oxytocin（OT） could induce cardiomyogenic differentiation of DFAT in vitro.METHODS： Mature adipocytes were isolated from human adipose tissue and were dedifferentiated into DFAT via ceiling culture.Cultured DFAT cells were treated with different concentrations of OT（1 × 10-8 mol / L,1 × 10-7 mol / L or 1 × 10-6 mol / L） for 1,2 or 3 weeks.DFAT cells cultured without OT served as control.Cells were evaluated morphologically and immunocytochemically as well as by RT-PCR and Western blot.Human cardiomyocytes were used as positive control.RESULTS： Most DFAT cells treated with 1 × 10-6 mol / L OT died after 3 days.After treatment with oxytocin（1 × 10-8 mol / L,1 × 10-7 mol / L） for 3 weeks,DFAT cells expressed cardiac markers including Nkx2.5,GATA4,and cardiac troponin T at both gene and protein levels.CONCLUSION： OT may induce cardiomyogenic differentiation of DFAT.
出处
《心脏杂志》
CAS
2011年第6期705-710,726,共7页
Chinese Heart Journal
关键词
去分化脂肪细胞
心肌分化
催产素
dedifferentiated FAT
cardiomyocyte differentiation
oxytocin
