Inhibition Of The Biosynthesis Of Oligosaccharides Prevents Cell Death Through The Introduction Of UPR

Cell death is an important life process and is the event that the creature ceases to play its function.This may be the result of a natural process in which old cells die and are replaced by new cells,or may be caused by factors such as disease,local damage,or the death of a cell-owned organism.Clinically,the development of many high-risk diseases is related to cell death,organ failure is often caused by large cell death,such as myocardial infarction,kidney failure and so on.At present,there is no means to prevent large cell death in clinic.Recent studies have shown that the activation of endoplasmic reticulum unfolded protein response is associated with myocardial protection.Here,we creatively propose a new mechanism for the protection of cardiac myocytes by inhibiting the biosynthesis of poly-sugars and subsequent N-glycosylation,activating unfolded protein response and inducing the expression of Grp94 and Grp78 to protect cells and inhibit death.The aim of this paper is to demonstrate a new mechanism for inhibiting cell death and protecting cells by inhibiting the synthesis of chitosan biosynthesis and the N-glycosylation of cell surface proteins,inducing protective unfolded protein response in endoplasmic reticulum.Firstly,it is proved that inhibition of poly-chitosan biosynthesis can protect cardiac myocytes and inhibit death.HSP90 inhibitor 17-aag(2?g/ml)was used to induce the apoptosis model of 16 h cells induced by NRK and h9c2 cells,and the model of cell necrosis was induced by TNF ?(20ng/ml)+z.vad.fmk and 30?m cell 40 h.At the same time,the cell model group of apoptosis necrosis was induced by the inhibitor glucosamine(5m M),2-deoxidation-glucose(1mM),TM(0.5?g/ml),2-fluoro-glucose(1mM).Using an inverted microscope to take a photographic record,LDH detected cytotoxicity and the activation of the associated apoptotic necrosis marker protein caspase-3 by Western blot,PARP cutting,RIP,and MLKLphosphorylation levels.It was found that the inhibitors of the biosynthesis of Chitosan had a significant protective effect on the cells that induce apoptotic necrosis.Secondly,it is proved that the protective effect of inhibiting the biosynthesis of chitosan is realized by activating denatured protein reaction and inducing the expression of endoplasmic reticulum companion protein.The NRK and H9C2 cells were given by using the chitosan biosynthesis inhibitor,and the phosphorylation-specific Western blot was used to prove the activation of the protein reaction factors such as ATF6,Perk and Ire1,and the expression of Grp94 and Grp78.To further prove that the protective effect of inhibiting the biosynthesis of poly-glucose is through the activation of unfolded protein response,we will use ATF6 specific inhibitors AEBSF,PERK specific inhibitors GSK2606414,Ire1-specific inhibitors such as 4?8c block the activation of denatured protein reactions and the expression of Grp94 and Grp78.The results showed that inhibiting the biosynthesis of poly-glycoprotein would activate all three factors of ATF6,perk and Ire1,and inhibit GSK2606414,4?8c activation by AEBSF,ATF6,perk or Ire1,Inhibits the expression of Grp94 and Grp78 induced by the biosynthesis inhibitor and weakens its protective effect.Through these experiments,we conclude that the protective effect of glucose is achieved by inducing the protective unfolded protein response in endoplasmic reticulum.