| OBJECTIVE This study aims to establish collagen-induced arthritis(CIA) rat model similar to RA pathogenesis, and compare different proteomic analysis of rats synovium among normal group, 25 days, 45 days CIA model group and paired Bizhongxiao Decoction (BZXD) treated group. It helps us identify proteins related to CIA rat pathological synovium, better understand pathogenesis of RA, as well as explore the molecular mechanism of Bizhongxiao Decoction (BZXD) which can be an effective therapy to CIA rats.METHODS CIA rat model was established. Bovine typeⅡcollagen (BIIC) and complete Freud adjuvant (CFA) were injected into the end of rats during the first immunization. They were injected second time after 21 days. Two weeks after first immunization, animals were evaluated according to their general condition, swelling of limbs and arthritis index (AI). Unsuccessfully established Animals were excluded. They were divided into Bizhongxiao Decoction (BZXD) treated group and model group randomly. BZXD treated group rats were administrated with BZX decoction, while model group rats were administrated with double distilled water. Normal group rats drink water freely. Synovial proteins were extracted, and two dimensional electrophoresis (2-DE) was used to separate total proteins from rat synovium of normal, 25 days, 45 days model and paired BZXD treated groups. Having been stained, proteins were analyzed by PDquest sottware. Some selected protein spots were identified by peptide mass fingerprint (PMF) based on Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and database.RESULTS CIA rat model was established successfully. 2-DE patterns with high resolution and reproducibility from rat synovial proteins was obtained. The average spots of rat synovial proteins from normal, 25 day model, 45 day model and paired BZXD treated groups were 1018±37, 1019±28, 1008±34, 1012±41, 1015±35. 19. Protein spots expressing differently were identified by PMF. These proteins are related to cell metabolism, signal transduction and so forth.CONCLUSION 1. We established CIA rat model and 2-DE protein profile of normal, 25 days, 45 days and paired BZXD group rat synovium. Proteins which express differently in 25 days and 45 days rat groups were obtained. This indicates that different expression of proteins exists in different stages in process of RA.2. Bizhongxiao decoction regulates multiple protein expression in different time point of CIA rat model. It is indicated that this decoction can be a therapy with various effects. |
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