Abstract
Diabetic nephropathy is considered one of the most common microvascular complications of diabetes and the pathophysiology involves multiple factors. Progressive diabetic nephropathy is believed to be related to the structure and function of the tubular epithelial cells in the kidney. However, the role of lysine acetylation in lesions of the renal tubular epithelial cells arising from hyperglycemia is poorly understood. Consequently, in this study, we cultured mouse renal tubular epithelial cells in vitro under high glucose conditions and analyzed the acetylation levels of proteins by liquid chromatography-high-resolution mass spectrometry. We identified 48 upregulated proteins and downregulated 86 proteins. In addition, we identified 113 sites with higher acetylation levels and 374 sites with lower acetylation levels. Subcellular localization analysis showed that the majority of the acetylated proteins were located in the mitochondria (43.17%), nucleus (28.57%) and cytoplasm (16.19%). Enrichment analysis indicated that these acetylated proteins are primarily associated with oxidative phosphorylation, the citrate cycle (TCA cycle), metabolic pathways and carbon metabolism. In addition, we used the MCODE plug-in and the cytoHubba plug-in in Cytoscape software to analyze the PPI network and displayed the first four most compact MOCDEs and the top 10 hub genes from the differentially expressed proteins between global and acetylated proteomes. Finally, we extracted 37 conserved motifs from 4915 acetylated peptides. Collectively, this comprehensive analysis of the proteome reveals novel insights into the role of lysine acetylation in tubular epithelial cells and may make a valuable contribution towards the identification of the pathological mechanisms of diabetic nephropathy.
Highlights
Diabetic nephropathy (DN) is regarded as one of the most common microvascular complications of diabetes and the pathophysiology involves multiple factors
Proteins were extracted from TKPT cells by radio immunoprecipitation assay (RIPA) lysis buffer (Beijing Soleibao Technology Co., Ltd) and protein concentrations were detected with a bicinchoninic acid (BCA) protein assay kit (Beijing Soleibao Technology Co., Ltd)
To further investigate the role of protein acetylation, the effect of high glucose on TKPT cells was determined by tandem mass tag (TMT) labeling technology and liquid chromatography-tandem mass spectrometry
Summary
Diabetic nephropathy (DN) is regarded as one of the most common microvascular complications of diabetes and the pathophysiology involves multiple factors. As one of the major pathological manifestations of diabetic nephropathy, renal tubulointerstitial fibrosis is strongly related to damage of the renal tubular epithelial cells arising from glucose, hypoxia, and inflammatory factors (Yang et al, 2020) It is well established from a variety of studies that the inhibition of apoptosis, pyroptosis, autophagy, and metabolic abnormalities may protect the structure and function of the renal tubular epithelial cells cultured with high glucose (Qin et al, 2018; Lv et al, 2019; Zhu et al, 2020). Protein acetylation is a post-translational modification (PTM) process in which acetyl groups are bound to amino acids by acetyltransferase This modification, which is common in eukaryotes and prokaryotes, is vital to the regulation of enzyme activity, protein stability and signaling pathways (Kouzarides, 2000). Much of the research conducted over recent years have shown that non-histone protein acetylation is involved in the regulation of the molecular biological function of cells (Narita et al, 2019)
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
