title: A novel cytoprotective peptide protects mesenchymal stem cells against mitochondrial dysfunction and apoptosis induced by starvation via Nrf2/Sirt3/FoxO3a pathway.
abstract: 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.
  
  
  
  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.
  
  
  
  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.
  
  
  
  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.
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, China.|Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.
authors.forename: Shuo
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, China.|Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, China.|Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.
authors.forename: Sidikejiang
authors.initials: S
authors.lastname: Niyazi
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, China.|Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.
authors.forename: Long
authors.initials: L
authors.lastname: Zheng
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, China.|Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.
authors.forename: Jiawei
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, China.|Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.
authors.forename: Weitao
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, China.|Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Xu
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, China.|Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.|Department of Transfusion, Zhongshan Hospital, Fudan University, Shanghai, China.
authors.forename: Ruiming
authors.initials: R
authors.lastname: Rong
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, China. esuperyc@163.com.|Shanghai Key Laboratory of Organ Transplantation, Shanghai, China. esuperyc@163.com.
authors.forename: Cheng
authors.initials: C
authors.lastname: Yang
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, 180 Fenglin Road, Shanghai, 200032, China. tyzhu@fudan.edu.cn.|Shanghai Key Laboratory of Organ Transplantation, Shanghai, China. tyzhu@fudan.edu.cn.|Shanghai Public Health Clinical Center, Fudan University, Shanghai, China. tyzhu@fudan.edu.cn.
authors.forename: Tongyu
authors.initials: T
authors.lastname: Zhu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D000066673:Cell Self Renewal / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D016895:Culture Media, Serum-Free; D019610:Cytoprotection / Q000187:drug effects; D000071316:Forkhead Box Protein O3 / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D008810:Mice, Inbred C57BL; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D051267:NF-E2-Related Factor 2 / Q000378:metabolism*; D010455:Peptides / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D056566:Sirtuin 3 / Q000378:metabolism*
chemical_list: D016895:Culture Media, Serum-Free; D000071316:Forkhead Box Protein O3; C474033:FoxO3 protein, mouse; D051267:NF-E2-Related Factor 2; C495636:Nfe2l2 protein, mouse; D010455:Peptides; D056566:Sirtuin 3
keywords: Cyclic helix B peptide; FoxO3a; Mesenchymal stem cell; Nrf2; Sirt3; Starvation