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©The Author(s) 2025. Published by Baishideng Publishing Group Inc. All rights reserved.
L-arginine from elder human mesenchymal stem cells induces angiogenesis and enhances therapeutic effects on ischemic heart diseases
Jian-Zhong Li, Xu Zhan, Hao-Bo Sun, Chao Chi, Guo-Fu Zhang, Dong-Hui Liu, Wen-Xi Zhang, Li-Hua Sun, Kai Kang
Jian-Zhong Li, Xu Zhan, Hao-Bo Sun, Chao Chi, Guo-Fu Zhang, Dong-Hui Liu, Wen-Xi Zhang, Kai Kang, Department of Cardiovascular Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin 150001, Heilongjiang Province, China
Jian-Zhong Li, Xu Zhan, Hao-Bo Sun, Kai Kang, Key Laboratory of Cell Transplantation of the National Ministry of Public Health, The First Affiliated Hospital of Harbin Medical University, Harbin 150001, Heilongjiang Province, China
Jian-Zhong Li, Department of Thoracic Surgery, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710014, Shaanxi Province, China
Li-Hua Sun, Department of Cardiovascular Surgery, The First Affiliated Hospital of Harbin Medical University and Pharmacology Department of Pharmacy College of Harbin Medical University, Harbin Medical University, Harbin 150081, Heilongjiang Province, China
Kai Kang, Key Laboratory of Hepatosplenic Surgery, Ministry of Education, Harbin 150001, Heilongjiang Province, China
Co-first authors: Jian-Zhong Li and Xu Zhan.
Co-corresponding authors: Li-Hua Sun and Kai Kang.
Author contributions: Li JZ and Zhan X contributed equally to this manuscript as co-first authors. Li JZ, Zhan X, and Sun HB performed the research and participated in data processing and statistical analyses; Chi C, Zhang GF, Liu DH, and Zhang WX provided experimental technical support; Li JZ, Sun LH, and Kang K wrote the manuscript and contributed to conception of the research project. Sun LH and Kang K contributed equally to this manuscript as co-corresponding authors.
Supported by the National Natural Science Foundation of China, No. 82472147; the Key Research and Development Program of Heilongjiang Province of China, No. 2023ZX06C04; and the Open Fund of Key Laboratory of Hepatosplenic Surgery, Ministry of Education, Harbin, China, No. GPKF202402.
Institutional review board statement: All human subject protocols were reviewed and approved by the Ethics Committee of the Second Affiliated Hospital of Harbin Medical University (Ethics No. 2014-R-010) and informed consent was obtained from each patient.
Institutional animal care and use committee statement: The animal experiments were conducted in accordance with the guidelines of the Institutional Animal Care and Use Committee of the Second Affiliated Hospital of Harbin Medical University, approval number 2015013.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Data sharing statement: sharing statement: All data generated or analyzed during this study are included in this published article and its supplementary information file.
Open Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
https://creativecommons.org/Licenses/by-nc/4.0/ Corresponding author: Kai Kang, Department of Cardiovascular Surgery, The First Affiliated Hospital of Harbin Medical University, No. 23 Youzheng Road, Harbin 150001, Heilongjiang Province, China.
kangkai1975@sina.com
Received: November 15, 2024
Revised: February 6, 2025
Accepted: March 17, 2025
Published online: April 26, 2025
Processing time: 159 Days and 12.5 Hours
BACKGROUND
Mesenchymal stem cell (MSC)-based therapy may be a future treatment for myocardial infarction (MI). However, few studies have assessed the therapeutic efficacy of adipose tissue-derived MSCs (ADSCs) obtained from elderly patients in comparison to that of bone marrow-derived MSCs (BMSCs) from the same elderly patients. The metabolomics results revealed a significantly higher L-arginine excretion from aged ADSCs vs BMSCs in hypoxic conditions. This was hypothesized as the possible mechanism that ADSCs showed an improved angiogenic capacity and enhanced the therapeutic effect on ischemic heart diseases.
AIM
To investigate the role of L-arginine in enhancing angiogenesis and cardiac protection by comparing ADSCs and BMSCs in hypoxic conditions for MI therapy.
METHODS
Metabolomic profiling of supernatants from ADSCs and BMSCs under hypoxic conditions were performed. Then, arginine succinate lyase (ASL) overexpression and short hairpin RNA plasmid were prepared and transfected into BMSCs. Subsequently, in vitro wound healing and Matrigel tube formation assays were used to verify the proangiogenetic effects of ADSC positive control, BMSCs, BMSCs ASL short hairpin RNA, BMSCs ASL overexpressed, and BMSC negative control on cocultured human umbilical vein endothelial cells. All sample sizes, which were determined to meet the statistical requirements and be greater than 3, were established on the basis of previously established literature standards. The protein levels of vascular endothelial growth factor (VEGF), basic fibroblast growth factor, etc. were detected. In vivo, the five types of cells were transplanted into the infarcted area of MI rat models, and the therapeutic effects of the transplanted cells were evaluated by echocardiography on cardiac function and by Masson’s staining/terminal-deoxynucleotidyl transferase mediated nick end labeling assay/immunofluorescence detection on the infarcted area.
RESULTS
Metabolomic analysis showed that L-arginine was increased. Using ASL gene transfection, we upregulated the production of L-arginine in aged patient-derived BMSCs in vitro, which in turn enhanced mitogen activated protein kinase and VEGF receptor 2 protein expression, VEGF and basic fibroblast growth factor secretion, and inductive angiogenesis to levels comparable to donor-matched ADSCs. After the cell transplantation in vivo, the modified BMSCs as well as ADSCs exhibited decreased apoptotic cells, enhanced vessel formation, reduced scar size, and improved cardiac function in the MI rat model. The therapeutic efficacy decreased by inhibiting L-arginine synthesis.
CONCLUSION
L-arginine is important for inducing therapeutic angiogenesis for ADSCs and BMSCs in hypoxic conditions. ADSCs have higher L-arginine secretion, which leads to better angiogenesis induction and cardiac protection. ADSC transplantation is a promising autologous cell therapy strategy in the context of the present aging society.
Core Tip: Adipose tissue-derived mesenchymal stem cells exhibited the advantages of angiogenesis compared to bone marrow-derived mesenchymal stem cells from elderly donors due to higher L-arginine secretion in serum-free and hypoxic conditions. In an animal model of MI, bone marrow-derived mesenchymal stem cells in which the arginine succinate lyase gene was integrated had a cardioprotective effect on angiogenesis. These results indicated that adipose tissue-derived mesenchymal stem cells from elderly patients with coronary heart disease and the introduction of L-arginine provide a promising strategy for stem cell therapy after MI.
