燕 松, 欧东波, 赵 佩, 李晓莉, 魏 婷, 陈 焱, 郑强荪. 甲状腺素促进miPS细胞向心肌细胞分化的非基因机制的作用[J]. 心脏杂志, 2016, 28(5): 518-522.
    引用本文: 燕 松, 欧东波, 赵 佩, 李晓莉, 魏 婷, 陈 焱, 郑强荪. 甲状腺素促进miPS细胞向心肌细胞分化的非基因机制的作用[J]. 心脏杂志, 2016, 28(5): 518-522.
    Effect of non-genomic mechanism of thyronine in promoting the differentiation of cardiomyocytes derived from mouse-induced pluripotent stem cells[J]. Chinese Heart Journal, 2016, 28(5): 518-522.
    Citation: Effect of non-genomic mechanism of thyronine in promoting the differentiation of cardiomyocytes derived from mouse-induced pluripotent stem cells[J]. Chinese Heart Journal, 2016, 28(5): 518-522.

    甲状腺素促进miPS细胞向心肌细胞分化的非基因机制的作用

    Effect of non-genomic mechanism of thyronine in promoting the differentiation of cardiomyocytes derived from mouse-induced pluripotent stem cells

    • 摘要: 目的 探讨三碘甲腺原氨酸(thyroxine,T3)在诱导小鼠诱导多能干细胞(miPSC)向心肌细胞分化过程中非基因的机制作用。方法 培养Oct4-GFP+ miPS细胞,利用悬滴培养形成拟胚体(embryoid body,EB)的方法诱导miPSC向心肌细胞分化,在诱导过程中分别添加甲状腺素T3和甲状腺素类似物三碘甲腺乙酸(triiodothyroacetic acid,Triac)。实验共分对照组、T3处理组、T3+Triac处理组。分别在诱导分化第4、6和12天,在倒置显微镜下观察细胞的分化情况,采用免疫荧光染色法检测分化第12天心肌细胞特异性结构蛋白α-actinin及肌钙蛋白(cardiac troponin,cTn)T的表达,Western blot法检测cTnT和p-JNK/JNK的表达情况。结果 诱导分化第4、6和12天,T3处理组跳动EB数量明显多于对照组和T3+Triac组(P<0.05);免疫荧光染色显示T3处理组的α-actinin和cTnT表达量较对照组和T3+Triac组明显增强(P<0.05);Western blot结果表明T3显著提高cTnT的表达量(P<0.05);细胞内信号分子JNK蛋白的磷酸化水平明显大于对照组和T3+Triac组(P<0.05)。结论 甲状腺素通过的非基因机制促进miPS细胞向心肌细胞分化,这种作用机制可能跟JNK信号通路有关。

       

      Abstract: AIM To investigate the non-genomic mechanism of thyroid hormone in the induction of miPS cells to differentiate into myocardial cells. METHODSWe cultivated miPS cells using the hanging drop method to culture and form embryoid bodies with thyroxine and triiodothyroacetic acid (which can competitively combine with thyroxine membrane receptor, Triac) under the same conditions to induce miPS cells to differentiate into myocardial-like cells. Ebs were cultured in three groups: control groups, T3 group and TH+Triac group. During different periods (on the 4th, 6th and 12th days) we observed the characteristics of differentiation under the inverted microscope. Immunohistochemical technique and Western blot method were used to detect interacting protein expression in different situations. RESULTSAccording to comparison of the three groups, in the thyroxine group not only the beating EB areas were greater than the control group and TH+Triac group, but miPS cells were also induced into myocardial-like cells, which were more powerful than in the control group. CONCLUSIONThe non-genomic mechanism of thyroid hormone infuences miPS cells to differentiate into myocardial cells, possibly via the JNKs signaling pathway.

       

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