Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran.
Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Tissue Cell. 2020 Dec;67:101429. doi: 10.1016/j.tice.2020.101429. Epub 2020 Aug 19.
Stem cell-based therapy has emerged as an attractive method for regenerating and repairing the lost heart organ. On other hand, poor survival and maintenance of the cells transferred into the damaged heart tissue are broadly accepted as serious barriers to enhance the efficacy of the regenerative therapy. For this reason, external factors, such as antioxidants are used as a favorite strategy by the investigators to improve the cell survival and retention properties. Therefore, the present study was conducted to investigate the In -vitro effect of L-carnitine (LC) on the telomere length and human telomerase reverse transcriptase (hTERT) gene expression in the cardiac differentiated bone marrow resident CD117 stem cells through Wnt3/β-catenin and ERK1/2 pathways. To do this, bone marrow resident CD117 stem cells were enriched by the magnetic-activated cell sorting (MACS) method, and were differentiated to the cardiac cells in the absence (-LC) and presence of the LC (+LC). Also, characterization of the enriched c-kit cells was performed using the flow cytometry and immunocytochemistry. At the end of the treatment period, the cells were subjected to the real-time PCR technique along with western blotting assay for measurement of the telomere length and assessment of mRNA and protein, respectively. The results showed that 0.2 mM LC caused the elongation of the telomere length and increased the hTERT gene expression in the cardiac differentiated CD117 stem cells. In addition, a significant increase was observed in the mRNA and protein expression of Wnt3, β-catenin and ERK1/2 as key components of these pathways. It can be concluded that the LC can increase the telomere length as an effective factor in increasing the cell survival and maintenance of the cardiac differentiated bone marrow resident CD117 stem cells via Wnt3/β-catenin and ERK1/2 signaling pathway components.
基于干细胞的疗法已成为一种有吸引力的方法，可用于再生和修复丧失的心脏器官。另一方面，转移到受损心脏组织中的细胞的存活率和维持率低，被广泛认为是增强再生治疗效果的严重障碍。出于这个原因，研究人员将抗氧化剂等外部因素作为首选策略，以提高细胞的存活率和保留特性。因此，本研究旨在通过 Wnt3/β-catenin 和 ERK1/2 通路研究左旋肉碱 (LC) 对心脏分化骨髓常驻 CD117 干细胞中端粒长度和人端粒酶逆转录酶 (hTERT) 基因表达的体外影响。为此，通过磁激活细胞分选 (MACS) 方法富集骨髓常驻 CD117 干细胞，并在不存在 (-LC) 和存在 LC (+LC) 的情况下将其分化为心脏细胞。此外，使用流式细胞术和免疫细胞化学对富集的 c-kit 细胞进行特征描述。在治疗期结束时，通过实时 PCR 技术和 Western blot 检测分别测量端粒长度和评估 mRNA 和蛋白。结果表明，0.2 mM LC 导致心脏分化 CD117 干细胞中端粒长度的延长和 hTERT 基因表达的增加。此外，这些通路的关键成分 Wnt3、β-catenin 和 ERK1/2 的 mRNA 和蛋白表达均显著增加。可以得出结论，LC 可以通过 Wnt3/β-catenin 和 ERK1/2 信号通路成分增加端粒长度，作为增加心脏分化骨髓常驻 CD117 干细胞存活率和维持的有效因素。
