Protective effects of total flavonoids from Alpinia officinarum on lead-induced HK-2 cell injury via Keap1/Nrf2/ARE signaling pathway
AIM To investigate the protective effects of total flavonoids from Alpinia officinarum Hance(TFAO)on lead-induced HK-2 cell injury.METHODS HK-2 cells cultured in vitro had their viability detected by MTT assay under the influence of TFAO or Pb.The control group,the TFAO control group,the model group,and the TFAO group were set up and exposed to 200 μmol/L Pb to induce cell injury in the groups except for the two control groups,meanwhile 100 μg/mL TFAO was given to the two TFAO groups for 24 h.After Pb-induction,the HK-2 cells had their apoptosis observed by Hoechst 33258 fluorescence staining combined with annexin V-FITC/PI double labeling flow cytometry;their intracellular ROS levels under Pb induction and TFAO intervention observed by fluorescence microscopy;their levels of ROS,MDA and GSH and the activities of GSH-Px,CAT and SOD determined by kit method;their levels of IL-1β,IL-6 and TNF-α in the supernatant of cell culture detected by ELISA;and their expressions of Keap1/Nrf2/HO-1 signaling pathway related proteins and caspase-3 protein detected by Western blot as well.RESULTS Compared with the control group,TFAO(12.5-200 μg/mL)intervention showed no significant effect on HK-2 cell viability.Compared with the model group,the TFAO group shared less apoptosis of Pb-induced HK-2 cells(P<0.05);alleviation of a series of apoptotic morphological changes including cell collapse,up-regulated activities of SOD,CAT and GSH-Px enzymes and GSH levels(P<0.05);lower levels of MDA and ROS(P<0.05);up-regulated expressions of Keap1/Nrf2/HO-1 signaling pathway related proteins and caspase-3 protein(P<0.05).CONCLUSION TFAO may play a protective role against Pb-induced HK-2 cell damage by regulating the expressions of proteins related to Keap1/Nrf2/ARE signaling pathway.
total flavonoids from Alpinia officinarum Hancelead poisoning,HK-2 cellsoxidative stressKeap1/Nrf2/ARE signaling pathway
国家科技期刊平台
NSTL
万方数据
维普
