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miR-221/222 Promote Endothelial Differentiation of Adipose-Derived Stem Cells by Regulation of PTEN/PI3K/AKT/mTOR Pathway

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Abstract

Adipose-derived stem cells (ADSCs) are a type of adult mesenchymal stem cell that show a repair effect on ischemic tissues owing to their capacity for endothelial differentiation. MicroRNA-221/222 (miR-221/222) has been extensively studied in endothelial cells (ECs). However, the mechanism that regulates ADSCs differentiation into ECs remains unknown. In this study, we investigated the effects of miR-221/222-overexpression/silence in ADSCs on endothelial differentiation by constructing lentiviral vectors. Differentiation capacity was assessed by measuring the expression of endothelial markers (CD31, CD34, and CD144). In addition, low-density lipoprotein (LDL) uptake and tube-like formation were performed for evaluation of functional characterization. The PTEN/PI3K/AKT/mTOR signaling pathway was investigated using western blotting to clarify the action mechanism of this gene. The revascularization of miR-221/222-transfeted ADSCs was further verified in a rat hind limb ischemia model. The results confirmed that transfection with miR-221/222 promoted the expression of endothelial markers, LDL uptake, and tube-like formation. As expected, the PI3K/AKT signaling pathway was effectively activated when ADSCs showed high expression of miR-221/222 during endothelial differentiation. Furthermore, injection of miR-221/222 transfected ADSCs significantly improved rat hindlimb ischemia, as evidenced by increased blood flow and structural integrity and reduce inflammatory infiltration. The results of this study suggest that miR-221/222 is essential for endothelial differentiation of ADSCs and provides a novel strategy for modulating vascular formation and ischemic tissue regeneration.

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Data Availability

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This work was supported by grants from the Natural Science Foundation of Anhui Province (2008085QH417 and 2108085QH333), the Natural Science Foundation of China (82103755), the Innovation program for Returned Overseas Chinese Scholars of Anhui Province (2021LCX027), and the College Students’ innovation and entrepreneurship training program (S202110367040 and 202210367034).

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Authors and Affiliations

  1. Anhui Engineering Technology Research Center of Biochemical Pharmaceutical, Department of Pharmacy, Bengbu Medical College, 2600 Donghai Avenue, Bengbu, 233030, China

    Wei Gao, Limin Yuan, Yue Zhang, Yue Si, Xuqing Wang, Tianci Lv & Yu-shuai Wang

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  1. Wei Gao
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Contributions

Guarantor of integrity of the entire study: Yu-shuai Wang; study concepts: Wei Gao; study design: Yu-shuai Wang; definition of intellectual content: Wei Gao; experimental studies: Wei Gao, Limin Yuan, and Yue Si; literature research: Yue Zhang; data analysis: Yue Si and Tianci Lv; manuscript preparation: Wei Gao; manuscript review: Yue Zhang.

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Correspondence to Yu-shuai Wang.

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All animal experimental procedures were conducted under the approval of the Institution Animal Ethical Committee of Bengbu Medical College (BBMC2019120).

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Gao, W., Yuan, L., Zhang, Y. et al. miR-221/222 Promote Endothelial Differentiation of Adipose-Derived Stem Cells by Regulation of PTEN/PI3K/AKT/mTOR Pathway. Appl Biochem Biotechnol 195, 4196–4214 (2023). https://doi.org/10.1007/s12010-023-04335-x

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