The Damaded Effect And Mechanism Of Glyoxal And Methylglyoxal On Human Embryonic Kidney Cells

Glyoxa(GO)and methylglyoxal(MG)are carbonyl compounds with high reactivity.GO and MG can be obtained through endogenous and exogenous pathways.When these compounds accumulate too much in the body,it will increase the level of reactive oxygen species(ROS)and lead to cell apoptosis eventually.These carbonyl compounds are closely related to a variety of diseases,including diabetic retinopathy,neuropathy and nephropathy,which affect human health.At present,the effect and mechanism of GO and MG on human embryonic kidney cells(HEK293)is still unknown.Therefore,it is of great significance to explore the specific mechanism of the damage effect of GO and MG on HEK293 cells.This study explored the damage effect and mechanism of GO and MG on HEK293 cells,and provided experimental evidence for elucidating its renal toxicity and preventing kidney damage.In this study,HEK293 cells were used to establish HEK293 cell models damaged by GO and MG.The cell viability was detected by MTT method,and the concentration of GO and MG used in subsequent experiments were chosen.DCFH-DA probe was used to detect intracellular ROS level.Hoechst33258 fluorescent staining experiment was used to detect cell apoptosis.Rhodamine 123(Rh123)fluorescent probe was used to detect changes in intracellular mitochondrial membrane potential.ELISA kit was used to detect 8-hydroxy-2 deoxyguanosine(8-OHdG)content.Western blot assay was used to detect the expression of key proteins in Mitogen-activated protein kinase(MAPK),c-Jun N-terminal kinase(JNK),nuclear factor kappa-B(NF-κB),ATM/Chk2/p53 signaling pathway,as well as the receptor of advanced glycation endproducts(RAGE),transforming growth factor-β1(TGF-β1)and apoptosis-related proteins.The results showed that 0.25-2 mM GO and MG could induce a significant decrease in HEK293 cell viability.0.5,1 and 2 mM GO and MG were used in subsequent experiments.0.5,1 and 2 mM GO and MG could increase intracellular ROS levels,reduce mitochondrial membrane potential,increase 8-OHdG content and promote cell apoptosis.The results of western blot showed that GO and MG significantly increased the ratio of Bax/Bcl-2,Chk2,p53,p21,RAGE and TGF-β1protein expression levels,promoted the phosphorylation of p38,JNK,IκB,ATM,H2 AX and activated p38 MAPK,JNK,NF-κB,ATM/Chk2/p53 signaling pathways.The above results indicated that GO and MG may induce HEK293 cell damage by regulating cell apoptosis,oxidative stress,inflammation and DNA damage.This study may provide experimental basis for elucidating the specific mechanism of GO and MG inducing HEK293 cells damage.