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Pentraxin 3 promotes cardiac differentiation of mouse embryonic stem cells through JNK signaling pathway

机译:五星素3通过JNK信号通路促进小鼠胚胎干细胞的心脏分化

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Abstract Cardiovascular disease is a leading cause of death worldwide, requiring the development of new therapeutic strategies including stem cell therapy. Pentraxins (PTXs) are a superfamily of proteins highly involved in different myocardial disorders, and thus this study aimed to identify the modulation of long pentraxin 3 (PTX3) in the differentiation of mouse embryonic stem cells (mESCs) toward cardiomyocytes. Cell toxicity of PTX3 was detected by MTT and LDH assays in mESCs. Embryoid bodies (EBs) were differentiated using hanging drop method, and the beating was observed under microscope. Expressional levels of early cardiac progenitor marker genes were assessed by qRT‐PCR. Expression of marker proteins in early myocardial development and the activation of JNK signaling pathway was evaluated by Western blot. PTX3 treatment at 50?ng/mL significantly promoted the expression of cardiac‐specific marker genes including Nkx2.5 , Mef2c , Tbx5 , dHand , and αMHC , and increased the expression of cardiac maturity indicative markers including connexin 43 and troponin C1. PTX3 enhanced the phosphorylation of JNK across the incubation duration, whereas the activation of p38 remained the same as control group. Co‐treatment of JNK signaling pathway inhibitor SP600125 impaired the PTX3‐promoted transcription of Nkx2.5 , Mef2c , Tbx5 , dHand , and αMHC . This study revealed the promotion of PTX3 in the differentiation of mESCs into cardiomyocytes and the underlying mechanism.
机译:摘要心血管疾病是全世界死亡的主要原因,需要开发新的治疗策略,包括干细胞疗法。五星素(PTX)是高度涉及不同心肌疾病的蛋白质的超家族,因此该研究旨在鉴定小鼠胚胎干细胞(MESCS)朝鲜细胞的长五花素3(PTX3)的调节。通过MESC中的MTT和LDH测定检测PTX3的细胞毒性。使用悬挂液滴法分化胚状体(EBS),在显微镜下观察到跳动。通过QRT-PCR评估早期心脏祖细胞标记基因的表达水平。通过蛋白质印迹评估标记蛋白在早期心肌发育中的表达及JNK信号传导途径的激活。 PTX3在50μg/ ml处的处理显着促进了存在NKX2.5,MEF2C,TBX5,DBAN和αmHC的心脏特异性标记基因的表达,并增加了包括Concexin 43和肌钙蛋白C1的心脏成熟度指示性标记的表达。 PTX3在孵育持续时间内增强了JNK的磷酸化,而P38的激活仍然与对照组相同。 JNK信号传导途径抑制剂SP600125的共同处理损害了NKX2.5,MEF2C,TBX5,DHAND和αMHC的PTX3促进的转录。本研究揭示了PTX3在麦塞斯分化到心肌细胞和潜在机制中的促进。

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