Study of the effect of keap1 on oxidative stress in human umbilical cord mesenchymal stem cells

作者全名："Deng, Hongrong; Chen, Yunxia; Liu, Huiwen; Wang, Li; Xu, Hao; Tan, Bin; Yi, Qin; Wang, Rui; He, Bolin; Tian, Jie; Zhu, Jing"

作者地址："[Deng, Hongrong; Chen, Yunxia; Liu, Huiwen; Wang, Li; Xu, Hao; Tan, Bin; Yi, Qin; Wang, Rui; Zhu, Jing] Chongqing Med Univ, Dept Pediat Res Inst, Natl Clin Res Ctr Child Hlth & Disorders, Minist Educ,Key Lab Child Dev & Disorders,Children, Chongqing, Peoples R China; [Xu, Hao] Chongqing Med Univ, Dept Clin Lab, Childrens Hosp, Chongqing, Peoples R China; [He, Bolin] Chongqing Med Univ, Childrens Hosp, Dept Blood Transfus, Chongqing, Peoples R China; [Tian, Jie] Chongqing Med Univ, Dept Cardiovasc Internal Med, Childrens Hosp, Chongqing, Peoples R China"

通信作者："Zhu, J (通讯作者)，Chongqing Med Univ, Dept Pediat Res Inst, Natl Clin Res Ctr Child Hlth & Disorders, Minist Educ,Key Lab Child Dev & Disorders,Children, Chongqing, Peoples R China."

来源：MOLECULAR BIOLOGY REPORTS

ESI学科分类：MOLECULAR BIOLOGY & GENETICS

WOS号：WOS:001134804300006

JCR分区：Q3

影响因子：2.6

年份：2024

卷号：51

期号：1

开始页：　

结束页：　

文献类型：Article

关键词：keap1; Oxidative stress; HucMSCs; IKK beta

摘要："Background HucMSCs had shown promising efficacy in treating childhood diseases, but oxidative stress induced by the poor microenvironment at the site of damage resulted in low cell survival after transplantation, thus preventing the cells from maximizing therapeutic efficacy. Therefore, this study aimed to investigate the role and mechanism of keap1 in oxidative stress injury of human umbilical cord mesenchymal stem cells (hucMSCs), and to provide theoretical support for improving the efficacy of stem cell therapy.Methods The hucMSCs were treated with hypoxic low-sugar-free serum (GSDH) to mimic the damaged site microenvironment after implantation. Adenoviral overexpression of keap1 gene of hucMSCs was performed in vitro, and cell proliferation ability was detected by CCK8 assay, crystal violet staining assay, and cell cycle assay. Cellular redox level was assessed by Amplex Red, MDA, and GSH/GSSG kit. Mitochondrial morphology was evaluated by mitotracker Red staining. ATP production was estimated by ATP detection kit. The mRNA and protein expression levels were tested by western blotting and RT-qPCR.Results GSDH treatment substantially upregulated keap1 expression. Subsequently, we found that overexpression of keap1 notably inhibited cell proliferation and caused cells to stagnate in G1 phase. At the same time, overexpression of keap1 induced the production of large amounts of H2O2 and the accumulation of MDA, but suppressed the GSH/GSSG ratio and the expression of antioxidant proteins NQO1 and SOD1, which caused oxidative stress damage. Overexpression of keap1 induced cells to produce a large number of dysfunctional mitochondria resulting in reduced ATP production. Moreover, Overexpression of keap1 significantly decreased the IKK beta protein level, while upregulating IkB mRNA levels and downregulating P50 mRNA levels.Conclusions Overexpression of keap1 may induce oxidative stress injury in hucMSCs by down-regulating IKK beta expression and inhibiting NF-kappa B pathway activation. This implies the importance of keap1 in hucMSCs and it may be a potential gene for genetic modification of hucMSCs."

基金机构：National Natural Science Foundation of China

基金资助正文：We are grateful to all authors who participated in this research.