Analysis of Serum Proteins and Enzymes Level in Human Subjects with Osteoarthritis

Abstract

Analysis of Serum Proteins and Enzymes Level in Human Subjects with OsteoarthritisThe aim of the present study was to assess the serum proteins and enzymes level using polyacrylamide gel electrophoretic (PAGE) profiles in human subjects with osteoarthritis (OA). Forty-one subjects with confirmed OA were selected for the present study. Sera were collected from these individuals and loaded in equal amounts on native and denaturing PAGE separately. Software analysis of these profiles was done using Scion Imaging (Beta release-4, Scion Corporation) and GelPro (Media Cybernetics, USA) programs. To visualize esterases (Est) and lactate dehydrogenase (LDH) isoenzymes in the sera of these patients substrate specific staining was performed. Differences in the values of control and OA subjects were tested statistically. Software analysis of native-PAGE profiles revealed the presence of nineteen peptides in control and twenty one in OA subjects respectively. Two extra peptides were present in the β-globulins region of OA subjects. Significant decline from 42.77% to 34.72% in albumin levels (hypoalbuminemia) was observed in OA subjects with total albumin to globulin ratio 0.58. In SDS-PAGE, the difference in control and OA subjects was observed among eight peptides with molecular weight 25, 22 and 20 kDa (absent in OA) and five novel peptides 270, 125, 30, 21.36 and 18.4 kDa (absent in controls), while albumin retains the major activity. For enzymes, Est follow a relative order, BchEst (42.86%)> ArylEst (16.24%)>AchEst (6.85%) in OA subjects with the expression of a new BchEst isoform in 4.78% and two isoforms of ArylEst at 2.13 and 1.61% concentrations respectively. Significantly declined albumin esterase-like activity (AlbEst) was observed (34%) (P<0.05) in diseased subjects compared with controls (47%). Significant increase in LDH-5 and decline in LDH-1 and -2 isoenzymes were also observed in the sera of OA subjects. However, the overall rank of LDH isoenzymes was similar in control and OA subjects. Our results demonstrate noticeable differences in the sera PAGE profiles and enzymes activity in control and OA subjects and provide evidence to select serum for its use in the search for suitable biochemical markers in osteoarthritis.