Abstract
To investigate proteins expressed in the renal tissue of the passive Heymann nephritis (pHN) rat model, we prepared pHN rat models with anti-FxA1 serum and analyzed the proteins differentially expressed in the kidney tissue with label-free liquid chromatography- tandem mass spectrometry. We then analyzed in depth the endoplasmic reticulum stress (ERS)-related protein using an online bioinformatics platform. Forty-one differential proteins and their annotations were obtained. Gene Ontology (GO) function analysis showed that 16 proteins were involved in cellular metabolism and 22 were proteins related to catalytic activity, including protein folding or ATPase. Protein-GO networks indicated that VCP could interact with the ERS marker HSPa5, with both involved in a single pathway. On inhibition of podocyte VCP by RNAi under normal conditions, the HSPa5 expression level did not change, but when the cell was subjected to ERS by tunicamycin, HSPa5 expression significantly increased with RNAi of VCP when compared with the tunicamycin-treated group. Our results showed that ERS plays an important role in podocyte injury of membranous nephropathy and is mediated by an HSPa5-VCP signaling pathway, in which the most predominant proteins are those related to cellular metabolism and catalytic activity.
Highlights
To investigate proteins expressed in the renal tissue of the passive Heymann nephritis rat model, we prepared pHN rat models with anti-FxA1 serum and analyzed the proteins differentially expressed in the kidney tissue with label-free liquid chromatography-tandem mass spectrometry
Gene Ontology (GO) function analysis showed that 16 proteins were involved in cellular metabolism and 22 were proteins related to catalytic activity, including protein folding or ATPase
Our results showed that endoplasmic reticulum stress (ERS) plays an important role in podocyte injury of membranous nephropathy and is mediated by an HSPa5-VCP signaling pathway, in which the most predominant proteins are those related to cellular metabolism and catalytic activity
Summary
To investigate proteins expressed in the renal tissue of the passive Heymann nephritis (pHN) rat model, we prepared pHN rat models with anti-FxA1 serum and analyzed the proteins differentially expressed in the kidney tissue with label-free liquid chromatography-tandem mass spectrometry. Our results showed that ERS plays an important role in podocyte injury of membranous nephropathy and is mediated by an HSPa5-VCP signaling pathway, in which the most predominant proteins are those related to cellular metabolism and catalytic activity. The current study analyzed differentially expressed proteins of the pHN rat model with high-throughput technology. Label-free quantification [7] MS can determine the differential expression of proteins In this method, proteins are compared based on the intensities of extracted ion chroma-. This study on the pHN rat model used label-free quantitative MS for high-throughput identification of differentially expressed proteins at several time points. The endoplasmic reticulum stress (ERS)-related protein was analyzed in depth using the online analysis platform found at http:// www.uniprot.org and http://bioinfo.capitalbio.com/mas3.0/
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