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Published on: July 13, 2018
A novel cytoprotective peptide protects mesenchymal stem cells against mitochondrial dysfunction and apoptosis
Shuo Wang1,2, Chao Zhang1,2, Sidikejiang Niyazi1,2
1Department of Urology, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, China.
Background:
Mesenchymal stem cell (MSC) has been widely explored in the past decade as a cell-based treatment for various diseases. However, poor survival of adaptively transferred MSCs limits their clinical therapeutic potentials, which is largely ascribed to the nutrient starvation. In this study, we determined whether a novel kidney protective peptide CHBP could protect MSCs against starvation and invested the underlying mechanisms.
Methods:
MSCs were subjected to serum deprivation and CHBP of graded concentrations was administered. Cell viability and apoptosis were detected by CCK-8, Annexin V/PI assay and Hoechst staining. ROS generation, mitochondrial membrane potential indicated by JC-1 and mitochondrial mass were measured by flow cytometry. The location of cytochrome c within cells was observed under fluorescence microscopy. Expressions of Nrf2, Sirt3, and FoxO3a were analyzed by western blot. In addition, preconditioning MSCs with CHBP was applied to test the possible protection against starvation. Finally, the effect of CHBP on the differentiation and self-renewal capacity of MSCs was also examined.
Results:
CHBP improved cell viability and suppressed apoptosis in a dose dependent manner. Starvation resulted in the mitochondrial dysfunction and treatment of CHBP could alleviate mitochondrial stress by diminishing oxidative injury of ROS, restoring mitochondrial membrane potential and maintaining mitochondrial membrane integrity. Importantly, Nrf2/Sirt3/FoxO3a pathway was activated by CHBP and Sirt3 knockdown partially abolished the protection of CHBP. Moreover, MSCs pretreated with CHBP were more resistant to starvation. Under normal condition, CHBP exerted little effects on the differential and self-renewal capacity of MSCs.
Conclusions:
The present study demonstrated the efficient protection of CHBP upon MSCs against starvation-induced mitochondrial dysfunction and apoptosis and indicated possible involvement of Nrf2/Sirt3/FoxO3a pathway in the protective effect.
Insights
A novel peptide, CHBP, enhances mesenchymal stem cell (MSC) survival against nutrient starvation by protecting mitochondria. This study reveals CHBP activates the Nrf2/Sirt3/FoxO3a pathway, improving MSC therapeutic potential.
Area of Science:
- Cell Biology
- Regenerative Medicine
- Biochemistry
Background:
- Mesenchymal stem cells (MSCs) show promise for treating diseases but suffer from poor survival due to nutrient starvation.
- This survival limitation hinders the clinical application of MSC-based therapies.
- Understanding mechanisms to enhance MSC resilience is crucial for advancing cell-based treatments.
Purpose of the Study:
- To investigate the protective effects of a novel kidney protective peptide, CHBP, on mesenchymal stem cells (MSCs) under nutrient starvation conditions.
- To elucidate the underlying molecular mechanisms responsible for CHBP-mediated protection of MSCs.
- To assess the impact of CHBP on MSC viability, apoptosis, and mitochondrial function during starvation.
Main Methods:
- MSCs were exposed to serum deprivation with varying concentrations of CHBP.
- Cell viability, apoptosis, reactive oxygen species (ROS) generation, and mitochondrial function were assessed.
- Key pathway proteins (Nrf2, Sirt3, FoxO3a) were analyzed using Western blot.
Main Results:
- CHBP significantly improved MSC viability and reduced apoptosis in a dose-dependent manner.
- CHBP alleviated starvation-induced mitochondrial dysfunction, including reducing ROS and restoring membrane potential.
- The protective effect of CHBP was linked to the activation of the Nrf2/Sirt3/FoxO3a pathway, with Sirt3 knockdown partially reversing the protection.
Conclusions:
- CHBP effectively protects mesenchymal stem cells against starvation-induced mitochondrial damage and apoptosis.
- The Nrf2/Sirt3/FoxO3a pathway plays a significant role in mediating CHBP's protective effects.
- CHBP holds potential for enhancing MSC survival and therapeutic efficacy in clinical applications.
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