氢和5-Aza通过p38 MAPK信号通路对脂肪源间充质干细胞成肌分化的协同作用。

IF 2.5 4区 医学 Q3 CELL & TISSUE ENGINEERING International journal of stem cells Pub Date : 2023-02-28 DOI:10.15283/ijsc21238
Wenyong Fei, Erkai Pang, Lei Hou, Jihang Dai, Mingsheng Liu, Xuanqi Wang, Bin Xie, Jingcheng Wang
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引用次数: 0

摘要

背景与目的:本研究旨在阐明氢与5-Aza在脂肪间充质干细胞(ADSCs)成肌分化中的调控机制和调控作用。方法与结果:在本研究中,ADSCs作为体外肌源性分化模式。首先,采用Alamar blue染色和线粒体示踪技术验证氢与5-Aza结合是否能促进细胞增殖。此外,本研究还通过Western blotting分析、细胞形态学特征分析(如肌管数目、长度、直径和成熟指数)、RT-PCR分析(Myod、Myogenin和Mhc mRNA表达)和免疫荧光分析(Desmin、Myosin和β-actin蛋白表达)来评估肌原性分化标志物(如Myogenin、Mhc和Myod蛋白表达)。最后,为了验证氢结合5-Aza的成肌分化机制,我们进行了生物信息学分析和Western blot检测p-P38蛋白的表达。氢与5-Aza结合,通过增加单细胞线粒体数量,上调Myod、Mhc和肌管形成等成肌生物标志物的表达,显著增强ADSCs体外增殖和成肌分化。氢与5-Aza结合可上调p-P38的表达。当细胞与SB203580 (p38 MAPK信号通路抑制剂)联合培养时,分化能力受到抑制。结论:氢可以减轻5-Aza的细胞毒性,并通过p38 MAPK通路协同促进脂肪干细胞的成肌分化能力。因此,上述结果提供了对肌源性分化的见解,并可能为骨骼肌相关疾病提供一种潜在的替代策略。
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Synergistic Effect of Hydrogen and 5-Aza on Myogenic Differentiation through the p38 MAPK Signaling Pathway in Adipose-Derived Mesenchymal Stem Cells.

Background and objectives: This study aims to clarify the systems underlying regulation and regulatory roles of hydrogen combined with 5-Aza in the myogenic differentiation of adipose mesenchymal stem cells (ADSCs).

Methods and results: In this study, ADSCs acted as an in vitro myogenic differentiating mode. First, the Alamar blue Staining and mitochondrial tracer technique were used to verify whether hydrogen combined with 5-Aza could promote cell proliferation. In addition, this study assessed myogenic differentiating markers (e.g., Myogenin, Mhc and Myod protein expressions) based on the Western blotting assay, analysis on cellular morphological characteristics (e.g., Myotube number, length, diameter and maturation index), RT-PCR (Myod, Myogenin and Mhc mRNA expression) and Immunofluorescence analysis (Desmin, Myosin and β-actin protein expression). Finally, to verify the mechanism of myogenic differentiation of hydrogen-bound 5-Aza, we performed bioinformatics analysis and Western blot to detect the expression of p-P38 protein. Hydrogen combined with 5-Aza significantly enhanced the proliferation and myogenic differentiation of ADSCs in vitro by increasing the number of single-cell mitochondria and upregulating the expression of myogenic biomarkers such as Myod, Mhc and myotube formation. The expressions of p-P38 was up-regulated by hydrogen combined with 5-Aza. The differentiating ability was suppressed when the cells were cultivated in combination with SB203580 (p38 MAPK signal pathway inhibitor).

Conclusions: Hydrogen alleviates the cytotoxicity of 5-Aza and synergistically promotes the myogenic differentiation capacity of adipose stem cells via the p38 MAPK pathway. Thus, the mentioned results present insights into myogenic differentiation and are likely to generate one potential alternative strategy for skeletal muscle related diseases.

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来源期刊
International journal of stem cells
International journal of stem cells Biochemistry, Genetics and Molecular Biology-Cell Biology
CiteScore
5.10
自引率
4.30%
发文量
38
期刊介绍: International Journal of Stem Cells (Int J Stem Cells), a peer-reviewed open access journal, principally aims to provide a forum for investigators in the field of stem cell biology to present their research findings and share their visions and opinions. Int J Stem Cells covers all aspects of stem cell biology including basic, clinical and translational research on genetics, biochemistry, and physiology of various types of stem cells including embryonic, adult and induced stem cells. Reports on epigenetics, genomics, proteomics, metabolomics of stem cells are welcome as well. Int J Stem Cells also publishes review articles, technical reports and treatise on ethical issues.
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