RESUMEN
Objective To screen for biomarkers in urine from patients with steroid-resistant nephrotic syndrome (SRNS) using surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) Proteinchip technology. Methods Urine samples from 9 SRNS patients, 32 steroid-sensitive nephrotie syndrome (SSNS) patients and 45 normal controls were analyzed using UA gold chip. Proteomic spectra were generated by mass spectrometry. Results Four differentially expressed biomarkers were identified with relative molecular weight of 6 703, 7 212, 11 820, 14 356. It was found that these protein peaks with relative molecular weigh of 7 212, 11 820, 14 356 were highly expressed in SRNS and 6 703 were lowly expressed in SRNS. The diagnostic cast that is constructed with these four protein to differentiate SRNS from SSNS with sensitivity of 88.89% and specificity of 93.75%. Conclusions SELDI-TOF-MS Proteinchip technology is a non-invasive, quick, easy, and convenient, and high-throughput analyzing method capable of screening several biomarkers from the urines of SRNS patients and has better clinical value.
RESUMEN
Proteomic analysis is an effective way to identify protein constituent in Lewy bedy-like inclusions (or aggresome) in vitro. Exposure to synthetic proteasome inhibitor (PSI, 10 μmol/L) for 48 hours was used to induce the formation of cytoplasmic proteineous inclusions (termed as PSi-induced inclusions) in PC12 cells.The proteomic approaches of biochemical fractionation, two-dimensional electrophoresis (2-D) and identification via peptide mass fingerprints (PMF) were deployed, and 20 protein components of LBs were identified,i ncluding 2 proteins involved in the production of synaptic neurotransmitter, 6 subunits of the 26 S proteasome,2 cytoskeleton proteins, 2 subunits of mitochondrial complexes, 1 anti-oxidant protein, and 7 chaperone proteins and (or) chaperone-like proteins. The results suggested that these LB protein components might had been recruited in PSI-induced inclusions formed in PC12 cells under the condition of proteasome inhibition.