催产素诱导去分化脂肪细胞向心肌细胞方向分化

杨 华; 陈连凤; 沈珠军

催产素诱导去分化脂肪细胞向心肌细胞方向分化

Oxytocin induces cardiac differentiation of dedifferentiated FAT cells

摘要

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(1×10-8 mol/L、1×10-7 mol/L、1×10-6 mol/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(1×10-6 mol/L)造成实验细胞大量死亡，无法进行后续试验。以1×10-8 mol/L及1×10-7 mol/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。

Abstract: 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.

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