【摘要】 背景:研究发现人和动物的多种间充质干细胞均存在河豚毒素敏感性钠离子通道,但钠离子通道是否会影响间充质干细胞向心肌细胞分化尚无共识。目的:观察钠离子通道特异性阻断剂河豚毒素对骨髓间充质干细胞向心肌样细胞分化的影响。方法:利用Percoll’s密度梯度离心法结合差速贴壁法分离大鼠骨髓间充质干细胞,用主动脉逆行生物酶灌流法分离大鼠成体心肌细胞。将DAPI标记的大鼠骨髓间充质干细胞与成体心肌细胞混合培养,应用0,10,100μmol/L的河豚毒素进行干预,1周后观察骨髓间充质干细胞向心肌样细胞的分化情况。结果与结论:免疫荧光细胞化学染色显示,大鼠骨髓间充质干细胞和成体心肌细胞混合培养1周后,骨髓间充质干细胞显著表达心肌特异性蛋白结蛋白和肌球蛋白,而10,100μmol/L的河豚毒素均可完全抑制结蛋白和肌球蛋白在骨髓间充质干细胞中的表达。提示钠离子通道是骨髓间充质干细胞向心肌样细胞分化的必要条件。更多还原

【Abstract】 BACKGROUND: The studies have found that the tetrodotoxin-sensitive sodium channels exist in various human and animal mesenchymal stem cells, but whether sodium channels may influence the differentiationof mesenchymal stem cells into myocardial cells has not been confirmed. OBJECTIVE: To observe the effect of sodium channel blocker tetrodotoxin on the differentiation of mesenchymal stem cells into cardiac-like cells. METHODS: Rat bone marrow mesenchymal stem cells were isolated with Percoll’s density gradient centrifugation and differential adhesion method, and then the adult rat myocardial cells were isolated by aorta retrograde collagenase perfusion; rat bone marrow mesenchymal stem cells labeled with 4’,6-diamidino-2-phenylindole were co-cultured with adult rat myocardial cells, and induced with 0, 10 and 100 μmol/L tetrodotoxin; and then the differentiation of bone marrow mesenchymal stem cells into cardiac-like cells was observed at 1 week after induction. RESULTS AND CONCLUSION: Immunofluorescence staining showed that the bone marrow mesenchymal stem cells could express cardiac-specific protein desmin and myosin after bone marrow mesenchymal stem cells co-cultured with myocardial cells for 1 week, while 10 and 100 μmol/L tetrodotoxin could inhibit the expressions of desmin and myosin in bone marrow mesenchymal stem cells. The results indicate that sodium channel may play a pivotal role in the differentiation of bone marrow mesenchymal stem cells into cardiac-like cells.更多还原