Objective To investigate the role and its mechanism of heat shock transcription factor 2(HSF2)in the development of podocyte pyroptosis and injury in diabetic kidney disease.Methods Twelve C57BL/6J male mice were randomly and equally divided into control and DKD groups,and immunohistochemical(IHC)staining was used to detect the expression of HSF2 in the kidney tissues of mice in the DKD group.In vitro culture of immortalized human podocytes was performed to construct a model of high glucose-induced podocyte injury,and the podocytes were divided into a low glucose group(LG group),a high glucose group(HG group),a hyperosmotic group(HO group),a high glucose overexpression control group(HG+OE-NC group),and a high glucose HSF2 overexpression group(HG+OE-HSF2 group),low glucose+knockdown control group(LG+si-NC group),low glucose+HSF2 knockdown group(LG+si-HSF2 group),blank control group(con group),knockdown control group(si-NCgroup),HSF2 knockdown group(si-HSF2 group),HSF2 knockdown+ROS inhibitor group(si-HSF2+MT group).Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR),protein blotting(Western blot)and immunofluorescence were used to detect the expression level of foot cell injury-related proteins synaptopodin and zo-1;Western blot was used to assess HSF2 protein in foot cells,NOD-like receptor protein 3(NLRP3)inflammatory vesicles and the expression levels of the focal death-associated proteins cleaved-Caspase-1 and IL-1 β proteins in foot cells were assessed by Western blotting;the mitochondrial superoxide red fluorescent probe kit was used to detect the mitochondrial superoxide(mtROS)levels in foot cells.Results Compared with the control group,HSF2 expression was significantly lower in the renal tissues of mice in the DKD group(P＜0.05).The relative expression levels of podocyte NLRP3,Cleaved-Caspase-1,IL-1β relative expression levels were higher than those in the LG group,HO group and HG+OE-HSF2 group,HSF2 expression levels were lower than those in the LG group and HO group,and the relative expression levels of synaptopodin,zo-1 protein and mRNA were lower than those in the LG group and HG+OE-HSF2 group(P＜0.05).Compared with the LG+si-NC group,the relative expression levels of foot cell synaptopodin,zo-1 protein and mRNA were significantly lower in the LG+si-HSF2 group,and the relative expression levels of NLRP3,Cleaved-Caspase-1,IL-1βrelative expression levels were significantly higher(P＜0.05).Compared with the LG group,mtROS of foot cells were significantly increased in the HG group and the LG+si-HSF2 group;compared with the HG group,mtROS of foot cells were significantly decreased in the HG+OE-HSF2 group;compared with the con group,mtROS of foot cells were significantly increased in the si-HSF2 group and the si-HSF2+MT groups both had significantly increased foot cell mtROS(P＜0.05).Conclusion HSF2 mediates high glucose-induced podocyte pyroptosis and injury by regulating the ROS/NLRP3 signalling pathway.
PodocytesHeat shock transcription factor 2cell pyroptosisNOD-like receptor protein 3 inflammasomeReactive oxygen species
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