| Objective:To observe the angiogenesis of joint synovial and lung lesions of collagen-induced arthritis with interstitial lung disease affected by Bitonglin pellet in rats, based on the concept from traditional Chinese medicine, to TCM arthralgia leather Bi syndrome endless, complex sense of the evil, the homes in the lung, limb Bi syndrome and lung Bi syndrome related theory, to observe angiogenesis on the collagen-induced arthritis joint synovial and lung lesions impacted by Bitongling pellet, and to provide experimental basis for clinical applications.Methods:1. In the currently accepted collagen typeⅡarthritis (CIA) model based on the use of bleomycin induced interstitial lung disease (ILD), explore the improvements in the established typeⅡcollagen-induced arthritis in rats and bleomycin induced model of interstitial lung disease combined (CIA-ILD).2. Observation with the naked eye on the overall status of each group in rats, activity behavior, joint swelling, respiratory conditions.3. Using reference to the literature for Assessment of arthritis scores (AI).4. Measuring two-foot volume changes with drainage act.5. Routine HE staining of the synovium of rats and lung histopathological changes, and analysis.6. Using FⅧfactor (factorⅧrelated antigen) antibody immunohistochemistry, synovial membranes were observed in lung tissue and microvessel density (MVD) change.7. Using real time quantitative PCR (Real-Time PCR, RT-PCR) detection method to observe the particles of each group of Bitongling synovial tissue and lung tissue of IL-17 mRNA, IL-1βmRNA, TNF-αmRNA,VEGFmRNA gene expression.8. Information and data to (?)±s,application of 2 -ΔΔCt method of real-time PCR data, when the target gene expression≥2 fold increased expression of the gene that, if that gene expression≤0.5 times lower. The remaining data applications using SPSS 17.0 statistical software for statistical analysis, the comparison between groups using analysis of variance to P<0.05 as statistically significant.Results:1. CIA-ILD experimental model of the overall state of the rats water consumption, food intake, coat gloss, less than the normal group activity behavior; joint swelling, and breathing accelerated more than the normal group. Department of the volume model group compared with the arthritis feet symptom score compared with the first 21 days of modeling, modeling the first 7 days showed significantly increase (P<0.05). Model group on day 7 between synovial thickening of the skin, inflammatory infiltration cells; modeling 14 days synovial mesothelium thickening, increased inflammatory cell infiltration; 21 days modeling mesothelial synovial thickening, inflammatory Infiltrating cells are more serious. Scores of synovial lesions for 21 days of modeling has increased more significantly than 7 days of modeling (P<0.05). The 7 day model group showed mild to moderate alveolar wall thickening, interstitial lymphocyte infiltration of plasma cells in inflammatory cells, local bleeding,14 days model showed severe alveolar wall thickening, interstitial lymphocyte infiltration of plasma cells,21 days model in lung tissue inflammatory cell infiltration showed patchy distribution, especially in the peribronchial lesions. Widened alveolar septum congestion, local proliferation of alveolar epithelial cells, some alveolar macrophages were seen in clusters of integrated group, fibroblasts, and fiber cell proliferation, pulmonary fibrosis alveolitis compared between models of 21 days increased more significantly than the 7 days model (P<0.05). Histopathology of the 7 days,14 days, and the first 21 days was observed, joint synovium and lung lesions appear and progress of successful modeling.2. Swelling of rat experimental model of arthritis of the treatment group has a different degree of joint swelling reduced arthritis scores were lower than the model group with Bitongling high, medium dose group and the effect of Tripterygium glycosides were significantly more (P<0.05). Model group with increased histological score of synovial, synovial tissue synovial cell proliferation, multi-storey or more, arranged the evacuation, disorder, inflammatory cell infiltration, and aggregation. Bitongling high dose group and middle dose group showed significant Tripterygium glycosides pathology improvements. The low dose group Bitongling also improved. Model can be seen in lung tissue inflammatory cell infiltration, and congestion widened alveolar septum alveolar hemorrhage and it showed fibrosis.3. Experimental model rats synovial tissue and lung issue showed relatively normal number of MVD increased in the Bitongling high and low dose group rats TwP synovial tissue and lung tissue compared with model group, the number of MVD reduction (P<0.05).4. Experimental model group showed IL-17mRNA expression in synovial≥2-fold than in the control group, where gene expression in rat synovial tissue and lung tissue increased. Bitongling high dose group showed IL-17mRNA expression in synovial≤1/2 times than in model group, where gene expression decreased. TwP group showed IL-17mRNA expression in the synovial fold≤1/2 than in the model group, where gene expression decreased. Model of lung tissue showed expression of IL-17mRNA≥2fold than the normal group, where gene expression increased. Bitongling high dose in lung tissue expression of IL-17mRNA showed to be≤1/2 times of the model group, where gene expression decreased. TwP in lung tissue expression of IL-17mRNA showed≤1/2 times of the model group, where gene expression decreased.5. Experimental model rats synovial tissue IL-1βmRNA expressed≥2-fold than the normal group, gene expression increased. Bitongling high-dose group and rats TwP synovial tissue IL-1βmRNA expressed≤1/2 times than the model group, the gene was down regulated. Model of lung tissue IL-1βmRNA expressed≥2-fold than the normal group, where the gene expression increased. The high dose group and Bitongling TwP lung tissue IL-1βmRNA expression decreased by≤1/2 fold than the model group down gene expression.6. Experimental model rats synovial tissue and lung tissue TNF-a mRNA expression≥2-fold than the normal group, gene expression increased. Bitongling high dose group and Tripterygium glycosides in rats synovial tissue expression of TNF-a mRNA≤1/2 times than the untreated group the gene was down regulated. Model group related lung TNF-a mRNA expression increased relatively to the normal group, high dose group and Bitongling Tripterygium glycosides in rat lung tissue showed TNF-a mRNA≤1/2 times than the untreated group where the gene was down regulated.7. Experimental model rats synovial tissue expression of VEGF mRNA relative to the normal group showed a≥2-fold increase in gene expression. High dose group Bitongling synovial tissue in model group showed≤1/2 times than in VEGF mRNA, the gene was down regulated. Model rats relative to normal lung tissue VEGF mRNA showed≥2 fold higher gene expression, Bitongling VEGF mRNA high dose group showed≤1/2 times than in model group lung tissue, the gene was down regulated. Tripterygium glycosides VEGF mRNA in the lung tissue showed≤1/2 times than in model group, gene expression decreased.Conclusion:1. In order to explore Bitongling particles joints of rheumatoid arthritis and lung disease, the CIA-ILD model was used for the first time. Histopathology 7 days,14 days, the first 21 days and the lung lesions appeared synovium and progress.2. Bitongling particles on arthralgia syndrome experimental model of rheumatoid arthritis synovial CIA-ILD and lung tissue in rats of two parts with inhibition of inflammatory pathology, high dose Jieyou efficacy, high dose group becomes more apparent.3. Bitongling pellet on the CIA-ILD rat synovial tissue and lung tissue pathology based on changes in inflammatory angiogenesis microvessel density (MVD) has significantly reduced the role of high dose are tabulated.4. Bitongling particles can down regulate four inflammatory cytokine gene expression in angiogenesis CIA-ILD synovial tissue and the lung tissue, and the role of high dose is significant.
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