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Ginsenoside Rg1 protects human umbilical cord blood-derived stromal cells against tert-Butyl hydroperoxide-induced apoptosis through Akt–FoxO3a–Bim signaling pathway

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Abstract

Human umbilical cord blood-derived stromal cells (hUCBDSCs) possess strong capability of supporting hematopoiesis and immune regulation, whereas some stress conditions cause reactive oxygen species (ROS) accumulation and then lead to oxidative injury and cell apoptosis. Ginsenoside Rg1 (G-Rg1) has been demonstrated to exert antioxidative and prosurvival effects in many cell types. In this study, the tert-Butyl hydroperoxide (t-BHP), an analog of hydroperoxide, was utilized to mimic the oxidative damage to hUCBDSCs. We aimed to investigate the effects of Ginsenoside Rg1 on protecting hUCBDSCs from t-BHP-induced oxidative injury and apoptosis, as well as the possible signaling pathway involved. It was shown that the treatment of hUCBDSCs with G-Rg1 markedly restored the t-BHP-induced cell viability loss, promoted the CFU-F formation, and inhibited cell apoptosis. G-Rg1 also caused a reduced production of LDH and MDA while significantly enhancing the activity of SOD. Mechanistically, G-Rg1 promoted the phosphorylation of Akt and FoxO3a and led to the cytoplasmic translocation of FoxO3a, which in turn suppressed FoxO3a-modulated expression of proapoptotic Bim and elevated the ratio of Bcl-2 to Bax. All these results suggest that G-Rg1 enhances the survival of t-BHP-induced hUCBDSCs and protects them against apoptosis at least partially through Akt–FoxO3a–Bim signaling pathway.

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Abbreviations

HIM:

Hematopoietic inductive microenvironment

HSCs:

Hematopoietic stem cells

G-Rg1:

Ginsenoside Rg1

t-BHP:

Tert-Butyl hydroperoxide

hUCBDSCs:

Human umbilical cord blood-derived stromal cells

BMSCs:

Bone marrow stromal cells

ROS:

Reactive oxygen species

PI3K:

Phosphoinositide-3-kinase

Akt:

Protein kinase B (PKB/Akt)

FoxO3a:

Forkhead box O3a

MDA:

Malondialdehyde

LDH:

Lactate dehydrogenase

SOD:

Superoxide dismutase

DMSO:

Dimethyl sulfoxide

CCK-8:

Cell counting kit-8

CFU-F:

Colony-forming unit of fibroblasts

7-AAD:

7-amino-actinomycin D

TUNEL:

Terminal dUTP nick end labeling

Br-dUTP:

Bromolated deoxyuridine triphosphate nucleotide

TdT:

Terminal deoxynucleotidyl transferase

PI:

Propidium iodide

DAPI:

4′, 6′-diamidino-2-phenylindole

PVDF:

Polyvinylidene difluoride

c-caspase 3:

Cleaved-caspase 3

Cyt c:

Cytochrome c

Apaf-1:

Apoptosis protease activating factor-1

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Acknowledgments

This work was financially supported by the National Natural Science Foundation of China (Grant number 30973818) and the National High-tech R&D Program of China (863 Program, Grant number 2014AA022302).

Author contributions

Liu Y. and Long Y. conceived and designed the experiments; Liu Y., Chen L.B., and Chen X.B. performed the experiments; Long Y. and Wang L. analyzed the data; Liu Y. prepared the manuscript; and Wang Y.P. supervised the whole research project.

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Authors and Affiliations

  1. Department of Histology and Embryology, Laboratory of Stem Cells and Tissue Engineering, Chongqing Medical University, Chongqing, People’s Republic of China

    Ying Liu, Lu Wang, Linbo Chen, ** Wang

  2. Research Center for Nutrition and Food Safety, Institute of Military Preventive Medicine, Third Military Medical University, Chongqing, People’s Republic of China

    Long Yi

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  1. Ying Liu
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Correspondence to Ya** Wang.

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Liu, Y., Yi, L., Wang, L. et al. Ginsenoside Rg1 protects human umbilical cord blood-derived stromal cells against tert-Butyl hydroperoxide-induced apoptosis through Akt–FoxO3a–Bim signaling pathway. Mol Cell Biochem 421, 75–87 (2016). https://doi.org/10.1007/s11010-016-2786-y

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