Effects of artesunate on hypoxia reoxygenation-induced HK-2 cell injury by regulating HMGB1-RAGE signaling pathway
CHEN Na
LI Lanmei
BAI Weiwei
SUN Shaoting
WANG Meng
ZHANG Ming
LI Yafen
Abstract：Objective To investigate the effect of artesunate(ART)on hypoxia/reoxygenation(H/R)-induced injury of human renal proximal tubular HK-2 cells by regulating the high mobility group protein B1(HMGB1)-receptor of advanced glycation endproduct(RAGE)signaling pathway.Methods HK-2 cells cultured in vitro were stochastically separated into control group,model group,L-ART group(4 μg/ml ART),H-ART group(16 μg/ml ART),H-ART+pcDNA NC group(16 μg/ml ART+transfected pcDNA NC plasmid),and H-ART+pcDNA-HMGB1 group(16 μg/ml ART+transfected pcDNA-HMGB1 plasmid).HK-2 cells in the control group were cultured normally,while the cells in the other groups were induced with hypoxia/reoxygenation(H/R).MTT and plate clone formation experiments were applied to detect the proliferation of cells in each group;ELISA kits were used to measure the expression of interleukin(IL)-1β,IL-6 and tumor necrosis factor-α(TNF-α)in various groups;flow cytometry was applied to detect the apoptosis in various groups;Western blot was applied to detect the expression of proliferation related proteins(PCNA),apoptosis related proteins(Bcl-2,Bax),and HMGB1-RAGE signaling pathway related proteins(HMGB1,RAGE)in each group.Results Compared with the control group,the survival rate,clone number,PCNA and Bcl-2 proteins expression of HK-2 cells decreased in the model group,but the apoptosis rate,IL-1β,IL-6,TNF-α,Bax,HMGB1 and RAGE proteins expression increased(P<0.05).Compared with the model group,the survival rate,clone number,PCNA and Bcl-2 proteins expression of HK-2 cells increased in L-ART and H-ART groups,but the apoptosis rate,IL-1β,IL-6,TNF-α,Bax,HMGB1 and RAGE proteins expression decreased(P<0.05).Compared with the H-ART and H-ART+pcDNA NC groups,the survival rate,clone number,PCNA and Bcl-2 proteins expression of HK-2 cells decreased in the H-ART+pcDNA-HMBB1 group,but the apoptosis rate,IL-1β,IL-6,TNF-α,Bax,HMGB1 and RAGE proteins expression increased(P<0.05).Conclusion ART may suppress inflammatory response and cell apoptosis,promote cell proliferation,and alleviate H/R-induced injury of renal proximal tubular HK-2 cells by inhibiting the HMGB1-RAGE signaling pathway.
Keywords：ArtesunateHigh mobility group protein B1-receptor of advanced glycation endproduct signaling pathwayHypoxia/reoxygenationHuman renal proximal tubule cellsCell injury
Publication Date：2024-11-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 812-817 )