ACAT1基因干扰对H2O2诱导的RPE细胞氧化应激模型糖代谢及细胞活性、迁移的影响机制OA

Objective To observe the effects of acetyl-CoA cholesterol acyltransferase 1(ACAT1)gene interference on glucose metabolism,cell viability and migration of hydrogen peroxide(H2O2)-induced oxida-tive stress model of retinal pigment epithelial(RPE)cells,and to explore the potential underlying mecha-nism.Methods Untreated normal human RPE cell line ARPE-19 cells were set as blank group.RPE cells on-ly induced by H2O2 served as induction group.H2O2-induced RPE cells transfected with ACAT1-siRNA were assigned to interference group.Six hours after ACAT1-siRNA transfection,5-adenosine monophosphate-acti-vated protein kinase(AMPK)pathway inhibitor compound C was added to H2O2-induced RPE cells for 6-hour treatment,which was defined as interference+compound C group.At 48 h after treatment,glucose-me-tabolism-related indicators were detected by commercial kits.MTT assay and Transwell assay were used to determine cell survival rate and migration capacity.Western blot was performed to detect the protein expres-sion levels of AMPK,p-AMPK,mammalian target of rapamycin(mTOR)and p-mTOR.Results Compared with the blank group,the induction group exhibited elevated adenosine triphosphate(ATP)level,glucose con-sumption and lactate production,as well as decreased cell survival rate and reduced number of transmembrane cells,with statistically significant differences(P<0.05).Compared with the induction group,the interference group showed increased ATP level,glucose consumption,lactate production,higher cell survival rate and more transmembrane cells(P<0.05).Compared with the interference group,the interference+compound C group presented decreased ATP level,glucose consumption,lactate production,lower cell survival rate and fewer transmembrane cells(P<0.05).The ratio of p-AMPK/AMPK was higher and p-mTOR/mTOR was lower in the induction group relative to the blank group(P<0.05).Compared with the induction group,the interfer-ence group had increased p-AMPK/AMPK(P<0.05)and decreased p-mTOR/mTOR(P<0.05).In con-trast to the interference group,the interference+compound C group displayed reduced p-AMPK/AMPK(P<0.05)and elevated p-mTOR/mTOR(P<0.05).Conclusion ACAT1 gene interference can promote glucose metabolism,cell viability and migration in H2O2-injured RPE cells,accompanied by alterations in the AMPK/mTOR signaling pathway.Compound C intervention can partially reverse these above effects,suggesting that this pathway may be involved in this process.However,rigorous genetic experiments are still required to fur-ther validate its exact mechanism.