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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。
BACKGROUND: It has been found that many kinds of mesenchymal stem cells of human and animals have tetrodotoxin-sensitive sodium channels, but whether sodium channels affect the differentiation of mesenchymal stem cells into cardiomyocytes is still unknown. Objective: To observe the effects of tetrodotoxin, a specific sodium channel blocker, on the differentiation of bone marrow-derived mesenchymal stem cells into cardiomyocytes. METHODS: Rat bone marrow mesenchymal stem cells were isolated by Percoll’s density gradient centrifugation combined with differential adherence method. Adult aortic cardiomyocytes were isolated by aortic cord blood perfusion. DAPI-labeled rat bone marrow mesenchymal stem cells were mixed with adult cardiomyocytes and treated with 0, 10 and 100μmol / L tetrodotoxin for one week. The differentiation of BMSCs into cardiomyocytes was observed after 1 week. RESULTS AND CONCLUSION: Immunofluorescence cytochemical staining showed that bone marrow mesenchymal stem cells express cardiac specific protein desmin and myosin at 1 week after mixed culture of rat bone marrow mesenchymal stem cells and adult cardiomyocytes, while 10,100 μmol / L tetrodotoxin can completely inhibit the expression of desmin and myosin in bone marrow mesenchymal stem cells. Tip sodium ion channels is a necessary condition for differentiation of bone marrow mesenchymal stem cells into cardiomyocyte-like cells.