L-carnitine cause to increase cell proliferation of C-Kit+ hematopoietic progenitor cells via decreasing the PI3K and FOXO-1 protein expression

Stem cell-based therapy has emerged as an attractive approach for regenerative medicine. Poor survival and maintenance of the cells used in regenerative medicine are considered as serious barriers to enhance the efficacy of the cell therapy. Using some antioxidants has been reported to prevent the a...

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Veröffentlicht in:Tissue & cell 2024-12, Vol.91, p.102558, Article 102558
Hauptverfasser: Valipour, Behnaz, Fathi, Ezzatollah, Farahzadi, Raheleh, Naderali, Elahe, Behniafar, Hamed
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Fathi, Ezzatollah
Farahzadi, Raheleh
Naderali, Elahe
Behniafar, Hamed
description Stem cell-based therapy has emerged as an attractive approach for regenerative medicine. Poor survival and maintenance of the cells used in regenerative medicine are considered as serious barriers to enhance the efficacy of the cell therapy. Using some antioxidants has been reported to prevent the aging of stem cells, and finding effective factors to reduce the senescence of these cells has impressive potential in cell therapy. The PI3K pathway adversely regulates the transcription factors known as FOXO, which are thought to have an inhibitory influence on cell proliferation. By downregulating FOXO and other targets, PI3K signaling controls the growth of cells. For this reason, the aim of the present study is to investigate the effect of L-carnitine (LC) as antioxidant on the cell proliferation and the protein expression of PI3K and FOXO. For understanding the in vitro effect of LC on the PI3K and FOXO-1 expression of C-kit+ hematopoietic progenitor cells, the bone marrow mononuclear cells were isolated, and C-kit+ cells was enriched by the magnetic-activated cell sorting (MACS). Next, the identification of enriched C-kit+ cells were done by flowcytometry and immunocytochemistry. Then, C-kit+ cells were treated with 0.2 mM LC, the cells were collected at the end of the treatment period (48 h), and the proteins were extracted. In the following, the protein expression of PI3K and FOXO-1 was measured by western blotting. In addition, flowcytometry was done to assess the Ki-67 expression as a key marker for cell proliferation investigation. 0.2 mM LC cause to significantly decrease in the protein expression of PI3K and FOXO-1 (*P
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Poor survival and maintenance of the cells used in regenerative medicine are considered as serious barriers to enhance the efficacy of the cell therapy. Using some antioxidants has been reported to prevent the aging of stem cells, and finding effective factors to reduce the senescence of these cells has impressive potential in cell therapy. The PI3K pathway adversely regulates the transcription factors known as FOXO, which are thought to have an inhibitory influence on cell proliferation. By downregulating FOXO and other targets, PI3K signaling controls the growth of cells. For this reason, the aim of the present study is to investigate the effect of L-carnitine (LC) as antioxidant on the cell proliferation and the protein expression of PI3K and FOXO. For understanding the in vitro effect of LC on the PI3K and FOXO-1 expression of C-kit+ hematopoietic progenitor cells, the bone marrow mononuclear cells were isolated, and C-kit+ cells was enriched by the magnetic-activated cell sorting (MACS). Next, the identification of enriched C-kit+ cells were done by flowcytometry and immunocytochemistry. Then, C-kit+ cells were treated with 0.2 mM LC, the cells were collected at the end of the treatment period (48 h), and the proteins were extracted. In the following, the protein expression of PI3K and FOXO-1 was measured by western blotting. In addition, flowcytometry was done to assess the Ki-67 expression as a key marker for cell proliferation investigation. 0.2 mM LC cause to significantly decrease in the protein expression of PI3K and FOXO-1 (*P&lt;0.05 and **P&lt;0.01, respectively). Also, the expression of Ki-67 was significantly increased in the presence of 0.2 mM LC (***P&lt;0.001). Briefly, LC can be considered an effective factor in increasing the proliferation of C-kit+ cells via some signaling pathways. •Aging causes reduced HSC function as well as self-renewal ability.•LC could be considered a suitable antioxidant for ameliorating the aging of C-kit+ HSCs.•The effect of LC on aging of C-kit+ HSCs via decreasing PI3K/FOXO protein as components of the aging-dependent pathway.</description><identifier>ISSN: 0040-8166</identifier><identifier>ISSN: 1532-3072</identifier><identifier>EISSN: 1532-3072</identifier><identifier>DOI: 10.1016/j.tice.2024.102558</identifier><identifier>PMID: 39260072</identifier><language>eng</language><publisher>Scotland: Elsevier Ltd</publisher><subject>Aging ; Animals ; C-kit+ hematopoietic progenitor cells ; Carnitine - pharmacology ; Cell Proliferation - drug effects ; Clinical agent ; Forkhead Box Protein O1 - metabolism ; Hematopoietic Stem Cells - cytology ; Hematopoietic Stem Cells - drug effects ; Hematopoietic Stem Cells - metabolism ; Phosphatidylinositol 3-Kinases - metabolism ; Proto-Oncogene Proteins c-kit - metabolism ; Regenerative medicine ; Signal Transduction - drug effects</subject><ispartof>Tissue &amp; cell, 2024-12, Vol.91, p.102558, Article 102558</ispartof><rights>2024 Elsevier Ltd</rights><rights>Copyright © 2024 Elsevier Ltd. 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For understanding the in vitro effect of LC on the PI3K and FOXO-1 expression of C-kit+ hematopoietic progenitor cells, the bone marrow mononuclear cells were isolated, and C-kit+ cells was enriched by the magnetic-activated cell sorting (MACS). Next, the identification of enriched C-kit+ cells were done by flowcytometry and immunocytochemistry. Then, C-kit+ cells were treated with 0.2 mM LC, the cells were collected at the end of the treatment period (48 h), and the proteins were extracted. In the following, the protein expression of PI3K and FOXO-1 was measured by western blotting. In addition, flowcytometry was done to assess the Ki-67 expression as a key marker for cell proliferation investigation. 0.2 mM LC cause to significantly decrease in the protein expression of PI3K and FOXO-1 (*P&lt;0.05 and **P&lt;0.01, respectively). Also, the expression of Ki-67 was significantly increased in the presence of 0.2 mM LC (***P&lt;0.001). 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subjects Aging
Animals
C-kit+ hematopoietic progenitor cells
Carnitine - pharmacology
Cell Proliferation - drug effects
Clinical agent
Forkhead Box Protein O1 - metabolism
Hematopoietic Stem Cells - cytology
Hematopoietic Stem Cells - drug effects
Hematopoietic Stem Cells - metabolism
Phosphatidylinositol 3-Kinases - metabolism
Proto-Oncogene Proteins c-kit - metabolism
Regenerative medicine
Signal Transduction - drug effects
title L-carnitine cause to increase cell proliferation of C-Kit+ hematopoietic progenitor cells via decreasing the PI3K and FOXO-1 protein expression
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