Piperazine ferulate attenuates high glucose-induced mesangial cell injury via the regulation of p66(Shc)

Diabetic nephropathy (DN) is a severe microvascular complication of diabetes. Hyperglycemia-induced glomerular mesangial cells injury is associated with microvascular damage, which is an important step in the development of DN. Piperazine ferulate (PF) has been reported to exert protective effects against the progression of DN. However, whether PF prevents high glucose (HG)-induced mesangial cell injury remains unknown. The aim of the present study was to investigate the effects of PF on HG-induced mesangial cell injury and to elucidate the underlying mechanisms. Protein and mRNA expression levels were determined via western blot analysis and reverse transcription-quantitative PCR, respectively. IL-6 and TNF-α levels were measured using ELISA. Reactive oxygen species levels and NF-κB p65 nuclear translation were determined via immunofluorescence analysis. Apoptosis was assessed by measuring lactate dehydrogenase (LDH) release, as well as using MTT and flow cytometric assays. The mitochondrial membrane potential of mesangial cells was determined using the JC-1 kit. The results revealed that LDH release were increased; however, cell viability and mitochondrial membrane potential were decreased in the HG group compared with the control group. These changes were inhibited after the mesangial cells were treated with PF. Moreover, PF significantly inhibited the HG-induced production of inflammatory cytokines and the activation of NF-κB in mesangial cells. PF also attenuated the HG-induced upregulation of the expression levels of fibronectin and collagen 4A1. Furthermore, the overexpression of p66(Src homology/collagen (Shc)) abolished the protective effect of PF on HG-induced mesangial cell injury. In vivo experiments revealed that PF inhibited the activation of inflammatory signaling pathways, glomerular cell apoptosis and mesangial matrix expansion in diabetic mice. Collectively, the present findings demonstrated that PF attenuated HG-induced mesangial cells injury by inhibiting p66(Shc).