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首页> 外文期刊>Cellular Physiology and Biochemistry >Puerarin Enhances Ca2+ Reuptake and Ca2+ Content of Sarcoplasmic Reticulum in Murine Embryonic Stem Cell-Derived Cardiomyocytes via Upregulation of SERCA2a
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Puerarin Enhances Ca2+ Reuptake and Ca2+ Content of Sarcoplasmic Reticulum in Murine Embryonic Stem Cell-Derived Cardiomyocytes via Upregulation of SERCA2a

机译：葛根素通过上调SERCA2a增强小鼠胚胎干细胞衍生心肌细胞中Ca2 +的再摄取和肌浆网的Ca2 +含量

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>Background/Aims: The embryonic stem cell-derived cardiomyocytes (ES-CMs) serve as potential sources for cardiac regenerative therapy. However, the immature sarcoplasmic reticulum (SR) function of ES-CMs prevents its application. In this report, we examined the effect of puerarin, an isoflavone compound, on SR function of murine ES-CMs. Methods: Murine ES-CMs were harvested by embryoid body-based differentiation method. Confocal calcium imaging and whole-cell patch clamps were performed to assess the function of SR. The mRNA expression levels of SR-related genes were examined by quantitative PCR. The protein expression of sarcoplasmic reticulum calcium-ATPase 2a (SERCA2a) was evaluated by immunofluorescent and western blot. Results: Long-term application of puerarin promotes basic properties of spontaneous calcium transient with increased amplitude, decay velocity, and decreased duration. Puerarin fails to alter ICa,L but increases the Ca2+ content of SR. Puerarin-treated ES-CMs have intact SR Ca2+ cycling with more SR Ca2+ reuptake. Long-term application of puerarin asynchronously upregulates the mRNA and protein expression of SERCA2a, as well as the transcripts of calsequestrin and triadin in developing ES-CMs. Application of puerarin during the stage of post-cardiac differentiation upregulates dose-dependently the transcripts of SERCA2a, phospholamban and tridin which can be reversed by the inhibitors of the PI3K/Akt and MAPK/ERK signaling pathways, but shows no effect on the protein expression of SERCA2a. Conclusion: This study demonstrates that long-term puerarin treatment enhances Ca2+ reuptake and Ca2+ content via upregulation of SERCA2a.
机译：> 背景/目标： 胚胎干细胞衍生的心肌细胞（ES-CM）可作为心脏再生疗法的潜在来源。但是，ES-CM的不成熟的肌浆网（SR）功能阻止了其应用。在本报告中，我们研究了异黄酮化合物葛根素对小鼠ES-CMs SR功能的影响。 方法： 通过基于类胚体的分化方法收获小鼠ES-CM。进行共聚焦钙成像和全细胞膜片钳以评估SR的功能。通过定量PCR检查SR相关基因的mRNA表达水平。通过免疫荧光和蛋白质印迹评估肌浆网钙ATPase 2a（SERCA2a）的蛋白表达。 结果： 长期使用葛根素可促进自发钙瞬变的基本特性，其幅度，衰减速度和持续时间均会增加。葛根素不能改变I Ca，L ，但会增加SR的Ca 2 + 含量。葛根素处理的ES-CM具有完整的SR Ca 2 + 循环，并且具有更多的SR Ca 2 + 重摄取。葛根素的长期应用在发育中的ES-CMs中异步上调SERCA2a的mRNA和蛋白表达以及calsequestrin和triadin的转录本。葛根素在心脏分化后阶段的应用可剂量依赖性地上调SERCA2a，phospholamban和tridin的转录本，这些转录本可被PI3K / Akt和MAPK / ERK信号通路的抑制剂逆转，但对蛋白质表达没有影响SERCA2a。 结论： 该研究表明，葛根素的长期治疗可通过以下途径增强Ca 2 + 的再摄取和Ca 2 + 的含量SERCA2a的上调。

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