Analyzing The Relative Expression Of Proteins Which Detected From The Ligament Of Hip Joint Of Ankylosing Spondylitis By Label-free Quantitative Technique

Ankylosing spondylitis (AS) is autoimmune disease characterized by chronic inflammation and abnormal ossification. At present, peripheral blood and synovial was the most common tissue in the research of AS because of the lesion tissue was hard to get and the knowledge on the pathogenesis of AS was rather limited. HPLC-MS/MS and label-free quantitation technology was used to analysis the proteomics of lesions from hip joint ligament.3 AS and 3 control samples were selected to study the pathogenesis of AS, especially the ossification mechanism. Comparing the protein data in our research with microarray data to screen some related proteins to the pathogenesis of AS.The MS data showed that there were 415 proteins only can detected in AS samples and 116 proteins only can detected in control samples, and 716 proteins can be detected in both. The proteins such as Collagen family, SLC93R1, HIST2H2AC, TNFRSF11B, ALPL, MMP13, TGFB1I1, ARL6IP5, COMP, TGFBI, CALM1, CALM2, MMP2, MMP3, CD44, SPTBN1 and CA1 et al were related to inflammation and ossification, this suggested that they may play an important role in AS.The result of label-free quantitation showed that there were 149 differential expression proteins and 84 were up-regulated,66 were down-regulated when P<0.5. There were 27 proteins in significant difference when the fold changes greater than 2 and 15 proteins up-regulated,12 proteins down-regulated among them. Compared to the microarray data, SLC9A3R1, KRT83, HIST2H2AC and RPL21 were up-regulated, PDLIM3, ARPC5, GLUD1, GDI2, VIM and ACTG1 were down-regulated.GO system showed that these proteins were involved in cell adhesion, Skeleton system structure, immune response, calcium ion bonding, angiogenesis and inflammation response et al. KEGG pathway analysis indicated that the related signal pathway involved in ossification, inflammation and bacterial invasion et al. String showed center node of the interactional network of differentially expressed proteins included COMP, ACTG1, FMOD and DCN, and the center node included COL2A1, COL5A1, SPARC, TIMP1 and LGALS3BP er al in the extracellular matrix protein.Finally, the experiment detected the proteins and analysis their related expression by HPLC-MS/MS and laber-free quantitation technology. Bioinformatics analysis were used to screen the key proteins related to the ossification mechanism of AS.