Piperazine ferulate attenuates high glucose‑induced mesangial cell injury via the regulation of p66Shc

Mol Med Rep. 2021 May;23(5):374. doi: 10.3892/mmr.2021.12013. Epub 2021 Mar 24.

Abstract

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 p66Src 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 p66Shc.

Keywords: piperazine ferulate; inflammatory cytokine; diabetic nephropathy; apoptosis; p66 Src homology/collagen; mesangial cells.

MeSH terms

  • Acute Kidney Injury / chemically induced
  • Acute Kidney Injury / drug therapy*
  • Acute Kidney Injury / genetics
  • Acute Kidney Injury / pathology
  • Animals
  • Collagen Type IV / genetics
  • Diabetic Nephropathies / drug therapy*
  • Diabetic Nephropathies / genetics
  • Diabetic Nephropathies / pathology
  • Disease Models, Animal
  • Fibronectins / genetics
  • Gene Expression Regulation / drug effects
  • Glucose / toxicity
  • Humans
  • Hyperglycemia / complications
  • Hyperglycemia / drug therapy
  • Hyperglycemia / genetics
  • Hyperglycemia / pathology
  • Interleukin-6 / genetics
  • Mesangial Cells / metabolism
  • Mesangial Cells / pathology
  • Mice
  • Piperazine / pharmacology*
  • RNA, Messenger / genetics
  • Repressor Proteins / genetics*
  • Transcription Factor RelA / genetics
  • Tumor Necrosis Factor-alpha / genetics

Substances

  • Col4a1 protein, mouse
  • Collagen Type IV
  • Fibronectins
  • GATAD2A protein, human
  • Interleukin-6
  • RNA, Messenger
  • Rela protein, mouse
  • Repressor Proteins
  • Transcription Factor RelA
  • Tumor Necrosis Factor-alpha
  • interleukin-6, mouse
  • Piperazine
  • Glucose

Grants and funding

The present study was supported financially by the National Natural Science Foundation of China (grant no. 81603171), Hunan Provincial Natural Scientific Foundation (grant nos. 2018JJ3743 and 2020JJ5841), Scientific Research Project of Hunan Provincial Health Commission (grant nos. B2019157 and B2019158) and the Open Sharing Fund for the Large-scale Instruments of Central South University (grant no. CSUZC202055).