Nicotiflorin Attenuates Cell Apoptosis In Renal Ischaemia-reperfusion Injury Through Activating Transcription Factor 3

Objective: Nicotiflorin is the main characteristic component of Nymphaea candida,which is a natural product that reportedly ameliorates acute injury of the liver caused by concanavalin A and galactosamine and cerebral cortex,but the effect of nicotiflorin on acute kidney injury(AKI)remains unknown.This study aimed to investigate the effects of nicotiflorin on ischemia/reperfusion(I/R)AKI and the associated mechanisms.Methods: We performed both(1)in vivo experiments with C57BL/6 mice with bilateral renal pedicles clamped for 45 min and(2)in vitro experiments with human kidney epithelial cells(HK-2)exposed to hypoxia/reoxygenation to mimic I/R injury to study the role of nicotiflorin in AKI.(3)Exploring the underlying mechanism of nicotiflorin acting kidney through computer simulation-molecular docking.Results: In vivo,nicotiflorin administration exerted protective effects on renal injury,as demonstrated by reductions in the levels of caspase3 and Bad(P <0.05),the upregulation of Bcl-2 expression(P <0.05)and improved renal histologic changes,which suggested that nicotiflorin can alleviate I/R injury and cell apoptosis.In vitro,nicotiflorin at a concentration of 75?μg/ml protected cells from hypoxia,which further confirmed that nicotiflorin exerts beneficial effects on hypoxia/reoxygenation.Through computational molecular docking,we found that activating transcription factor 3(ATF3)exhibits a robust interaction with nicotiflorin with a simulated binding energy of-9.2 degrees.We confirmed that the effect of nicotiflorin was weakened by the knockdown of ATF3,verified the interaction of nicotiflorin with ATF3 in HK-2 cells,and found that nicotiflorin reduced the apoptosis of renal tubular epithelial cells through ATF3.Conclusion: Based on the above-described results,nicotiflorin appears to have a beneficial impact on deteriorated renal function,as demonstrated using an experimental I/R model.The underlying mechanisms of nicotiflorin might inhibit HK-2 cell apoptosis through ATF3.