Effect of hyperoside on glomerular endothelial cell damage induced by high glucose in rats based on the TXNIP/NLRP3 pathway

YIN Bo; XIONG Yuling; PENG Linlin

doi:10.13431/j.cnki.immunol.j.20250122

Abstract：Objective To investigate the effect of hyperoside(Hyp)against high glucose-induced injury in rat glomerular endothelial cells(RGECs)based on the thioredoxin-interacting protein(TXNIP)/NOD-like receptor pyrin domain-containing protein 3(NLRP3)pathway.Methods Rat RGECs with a fusion degree of 70%to 80%were selected and divided into the normal group(cultured in a 5.5 mmol/L glucose medium),the high glucose group(cultured in a 30 mmol/L glucose medium),the Hyp-low group(30 mmol/L glucose+100 μg/mL Hyp treatment),the Hyp-high group(30 mmol/L glucose+400 μg/mL Hyp treatment),the pcDNA group(transfected with pcDNA plasmid+30 mmol/L glucose medium culture),the pcDNA-TXNIP group(transfected with pcDNA-TXNIP plasmid+30 mmol/L glucose medium culture),the Hyp-high+pcDNA group(transfected with pcDNA plasmid+30 mmol/L glucose+400 μg/mL Hyp treatment),and the Hyp-high+pcDNA-TXNIP group(transfected with pcDNA-TXNIP plasmid+30 mmol/L glucose+400 μg/mL Hyp treatment).5-Bromo-2'-deoxyuridine(BrdU)and MTT were used to detect the proliferation of rRGECs,and flow cytometry was employed to examine the apoptosis of rat RGECs.The levels of malondialdehyde(MDA),nitric oxide(NO),tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)in rat RGECs were measured.FITC-polyethylene glycol was used to assess the permeability of rat RGECs,and real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-qPCR)and Western blot were used to detect the mRNA and protein expression levels of TXNIP and NLRP3 in rat RGECs.Results The optical density(OD),BrdU positive ratio,and NO in the high glucose group were lower than those in the normal group(P<0.05).The OD,BrdU positive ratio,and NO were higher in the Hyp-low group and Hyp-high group than in the high glucose group(P<0.05),and higher in the Hyp-high group than in the Hyp-low group(P<0.05).The OD,BrdU positive ratio,and NO were lower in the pcDNA-TXNIP group than in the pcDNA group(P<0.05),and lower in the Hyp-high+pcDNA-TXNIP group than in the Hyp-high+pcDNA group(P<0.05).The cell apoptosis rate,permeability,MDA,TNF-α,IL-6 levels,and TXNIP,NLRP3 mRNA,and protein expression in the high glucose group were higher than those in the normal group(P<0.05),which,however,were lower in the Hyp-low group and Hyp-high group than in the high glucose group(P<0.05).The cell apoptosis rate,permeability,MDA,TNF-α,IL-6 levels,and TXNIP,NLRP3 mRNA,and protein expression in the Hyp-high group were lower than those in the Hyp-low group(P<0.05),which were higher in the pcDNA-TXNIP group than in the pcDNA group(P<0.05),and higher in the Hyp-high+pcDNA-TXNIP group than in the Hyp-high+pcDNA group(P<0.05).Conclusion Hyp reduces high glucose-induced rat RGECs by inhibiting the TXNIP/NLRP3 pathway.