iTRAQ‐Based Proteomics of Chronic Renal Failure Rats after FuShengong Decoction Treatment Reveals Haptoglobin and Alpha‐1‐Antitrypsin as Potential Biomarkers

Abstract

Background. Chronic renal failure (CRF) has become a global health problem and bears a huge economic burden. FuShengong Decoction (FSGD) as traditional Chinese medicine has multiple pharmacological effects. Objectives. To understand the underlying molecular mechanism and signaling pathway involved in the FSGD treatment of CRF and screen differentially expressed proteins in rats with CRF treated with FSGD. Methods. Thirty-three male Sprague-Dawley rats were randomly divided into control group, CRF group, and FSGD group. Differentially expressed proteins were screened by iTRAQ coupled with nanoLC-MS/MS, and these identified proteins were later analyzed by GO, KEGG, and STRING. Additionally, haptoglobin (HP) and alpha-1-antitrypsin (AAT) were finally verified by ELISA, Western blot, and real time PCR. Results. A total of 417 proteins were identified. Nineteen differentially expressed proteins were identified in the FSGD group compared with the model group, of which 3 proteins were upregulated and 16 proteins were downregulated. Cluster analysis indicated that inflammatory response was associated with these proteins and complement and coagulation cascade pathways were predominantly involved. The validation methods further confirmed that the levels of HP and AAT were significantly increased. Conclusions. HP and AAT may be the important biomarkers in the pathogenesis of CRF and FSGD therapy.

Urine proteomics of primary membranous nephropathy using nanoscale liquid chromatography tandem mass spectrometry analysis

“…Due to its anti-protease activity, A1AT can exert tissue-protective effects. A1AT levels are upregulated in the renal tissue of adenine-induced chronic renal failure model [ 34 ] and in vivo administration of clinical grade A1AT improves renal function, decreases acute tubular necrosis and ameliorates acute kidney injury following experimental kidney ischemia reperfusion damage [ 35 ]. The overexcretion of A1AT have been described in the urine of IgA nephropathy, membranoproliferative glomerulonephritis, minimal change disease, focal segmental glomerulosclerosis and membranous nephropathy [ 36 , 37 ].…”

“…The overexcretion of A1AT have been described in the urine of IgA nephropathy, membranoproliferative glomerulonephritis, minimal change disease, focal segmental glomerulosclerosis and membranous nephropathy [ 36 , 37 ]. Upregulation of A1AT could lead to inhibition of neutrophil elastase, which can contribute to accumulation of mesangial matrix, maintaining the elasticity of blood vessels and glomerular integrity [ 34 ]. A1AT is located in the cytoplasm of podocytes within sclerotic glomeruli [ 13 ] and its relationship with podocyte dysfunction and podocyte stress need to be further investigated.…”

Effects of Long Non-Coding RNA LINC00963 on Renal Interstitial Fibrosis and Oxidative Stress of Rats with Chronic Renal Failure via the Foxo Signaling Pathway

“…Ten rats (five rats in each group) were sacrificed. The criteria of the successful establishment [19] was as follows: the appetite reduction, loss of weight, increased water inflow, elevated urine volume, (3.3 ± 0.5) ml/rat, and increased urine protein (5.0 ± 0.5) mg/rat were found in the CRF rats. The rat scarification method was as follows: rats were intraperitoneally injected with 2% sodium pentobarbital and fixed in the supination position after anesthesia.…”

Complement factors and alpha‐fetoprotein as biomarkers for noninvasive prenatal diagnosis of neural tube defects

“…Serum pools were depleted of high-abundance proteins using the Human 14 Multiple Affinity Removal System (Agilent Technologies) as described and reported in our previous studies. 23,24 After desalination and concentration of low abundance components, SDT Lysis buffer (4% SDS (161-0302, Bio-Rad), 100 mM Tris (A6141, Sigma)-HCl (10011018, Sinopharm), 1 mM DTT (161-0404, Bio-Rad), pH 7.6) was added to the sample, which was then boiled for 15 minutes. After centrifuging at 14,000 × g for 40 min, the supernatant protein was quantified with the BCA Protein Assay Kit (Bio Rad, Hercules, CA).…”

“…22 Isobaric tags for relative and absolute quantitation (iTRAQ) has become one of the main tools used in quantitative proteomic research. Because of its high sensitivity, strong separation ability, wide analysis range, and more reliable quantitative results, 23 iTRAQ is suitable for identifying molecular biomarkers of genetic diseases with complex changes, including NTDs. In early 2010, Kolla et al used iTRAQ to search for novel biomarkers of Down syndrome (DS) in maternal plasma and identified numerous elevated proteins, including serum amyloid P component, amyloid beta A4, gamma-actin, and titin in DS pregnancies.…”