Dickkopf相关蛋白3对高糖诱导的新生大鼠心肌细胞损伤的影响OA

Aim:To investigate the effects of Dickkopf-3(DKK3)on high glucose-induced injury of neonatal rat car-diomyocyte(NRCM).Methods:NRCM was isolated and cultured.Cells were transfected with Ad-DKK3 to establish a DKK3 overexpression model(Ad-NC transfection was used as a control),and then were stimulated with 30 mmol/L glucose to establish a high glucose injury model(5.5 mmol/L glucose was used as a control),and named as control group,DKK3 overexpression group,high glucose group,and high glucose+DKK3 overexpression group.After 24 hours incubation,cell ac-tivity was assessed by MTT assay.IL-1β,IL-6,TNF-α mRNA expressions in NRCM were measured using qRT-PCR.Malon-dialdehyde(MDA)content,NADPH oxidase activity,and superoxide dismutase(SOD)activity in NRCM were determined using colorimetric assay.Protein expressions of DKK3,NF-κB,Nrf2,Wnt,β-catenin,and c-Myc in NRCM were detected by Western blot.Co-immunoprecipitation was used to detect the binding of DKK3 to Wnt pathway receptors.Results:High glu-cose stimulation significantly downregulated DKK3 expression,reduced cell activity,increased IL-1β,IL-6,TNF-α mRNA expression levels,elevated MDA content and NADPH oxidase activity in cell culture supernate,decreased SOD activity,up-regulated the expressions of NF-κB,Wnt,β-catenin,and c-Myc proteins and downregulated the expression of Nrf2;while DKK3 overexpression significantly reversed above effects(P<0.05).A significant interaction was found between high glu-cose and DKK3 overexpression(P<0.05).DKK3 directly bound to the Wnt pathway receptors.Conclusion:DKK3 could alleviate NRCM injury caused by high glucose.The mechanism lies in its ability to bind to Wnt pathway receptors and inhib-it the activity of Wnt pathway.