The Function And Mechanism Of Reactive Oxygen Species In The Nucleus In The Process Of Somatic Cell Reprogramming

Somatic cell reprogramming is the process of reversing highly differentiated somatic cells back to pluripotent stem cells through specific technical methods.Since its introduction in 2006,this technology has gained significant attention and research due to its enormous potential in regenerative medicine,toxicology screening,and in vitro disease modeling.So far,various methods of somatic cell reprogramming have been reported,but many of the mechanisms involved in the process are still unknown.In order to apply reprogramming technology in clinical settings,it is necessary to understand the pluripotency regulatory network and signal transduction mechanisms involved.Reactive oxygen species(ROS)is a collection of compounds,including superoxide anion,hydrogen peroxide,hydroxyl radical,peroxynitrite that are byproducts of cellular metabolism,mainly generated in mitochondria.Traditionally,ROS was considered a class of harmful substances produced by cells that could cause oxidative damage to proteins,RNA,DNA,and other organelle in cells.However,ROS in cells also have unique functions and play an important role in signal transduction processes have been shown in recent studies.It is now established that the overall level of ROS in cells has a direct impact on the process of somatic cell reprogramming,but the function of ROS within the cell nucleus on somatic cell reprogramming has not yet been reported.The main focus of this study is to investigate the mechanism of the role of ROS within the cell nucleus in somatic cell reprogramming.Firstly,we used a protein oxidation level detection kit to measure the oxidation level of nuclear proteins during the early stages of somatic cell reprogramming and found that it was increased.We then used our own modified nuclear-localized ROS probe,which can indicate ROS levels in real-time,and confirmed an increase in ROS levels within the cell nucleus during the early stages of somatic cell reprogramming through confocal imaging analysis.We also specifically expressed enzymes and proteins from the redox system within the cell nucleus and found that overexpressing Trx1(thioredoxin)can reduce ROS levels within the cell nucleus and promote reprogramming,proving that ROS within the cell nucleus during somatic cell reprogramming is not conducive to reprogramming.Furthermore,this study also discovered that nuclear-localized Trx1 can enhance the nuclear accumulation of HDAC7 and promote MET,thereby improving reprogramming efficiency.In summary,this paper’s research results indicate that ROS in the cell nucleus increases in the early stages of reprogramming and is detrimental to the process.Lowering ROS levels in the cell nucleus can promote reprogramming efficiency.This study supplements the understanding of the distribution and function of ROS in cells,as well as the metabolic pathways and signal transduction mechanisms involved in somatic cell reprogramming.It was revealed that changes in ROS at the subcellular level can affect MET by influencing the redox state of HDAC7,thus improving reprogramming efficiency.