L-carnitine Extends the Telomere Length of the Cardiac Differentiated CD117+- Expressing Stem Cells
L-carnitine Extends the Telomere Length of the Cardiac Differentiated CD117+- Expressing Stem Cells
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DOI：
10.1016/j.tice.2020.101429
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发表时间：
2020-12-01
期刊：
影响因子：
2.6
通讯作者：
Vietor, Ilja
中科院分区：
文献类型：
--
作者：
Fathi, Ezzatollah;Farahzadi, Raheleh;Vietor, Ilja
基于干细胞的治疗已经成为再生和修复失去的心脏器官的一种有吸引力的方法。另一方面，转移到受损心脏组织中的细胞的存活和维持差被广泛接受为增强再生治疗功效的严重障碍。出于这个原因，外部因素，如抗氧化剂，被研究人员用作改善细胞存活和保留特性的最佳策略。因此，本研究通过Wnt 3/beta-catenin和ERK 1/2途径，探讨左旋卡尼汀（L-carnitine，LC）对体外培养的心肌分化骨髓CD 117（+）干细胞端粒长度和人端粒酶逆转录酶（human telomerase reverse transcriptase，hTERT）基因表达的影响。为此，通过磁激活细胞分选（MACS）方法富集骨髓驻留的CD 117（+）干细胞，并在不存在（-LC）和存在LC（+LC）的情况下分化为心肌细胞。此外，使用流式细胞术和免疫细胞化学对富集的c-kit（+）细胞进行表征。在处理期结束时，对细胞进行实时PCR技术沿着蛋白质印迹分析，以分别测量端粒长度并评估mRNA和蛋白质。结果表明，0.2mM LC可使心肌分化的CD 117（+）干细胞端粒长度延长，hTERT基因表达增加。此外，作为这些通路的关键组分，Wnt 3、β-连环蛋白和ERK 1/2的mRNA和蛋白表达显著增加。结论：LC可通过Wnt 3/beta-catenin和ERK 1/2信号通路增加心肌分化骨髓CD 117（+）干细胞的端粒长度，从而提高心肌分化骨髓CD 117（+）干细胞的存活和维持。
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/beta-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, beta-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/beta-catenin and ERK1/2 signaling pathway components.