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Objective: To establish rat vascular endothelial growth factor in real-time quantitative PCR method, and type Ⅱ collagen -induced arthritis rat model, evaluation Tripterygium glycosides on VEGF mRNA expression levels in different tissues , and to explore its therapeutic rheumatoid arthritis may be the mechanism . Method : 1. Cross intron primers and TaqMan probe , species-specific authentication , the cycler ABI7500 create real-time fluorescence detection method , amplified fragments TA cloning , sequencing and production standards , analysis of its specificity, sensitivity and repeatability . 2 rat type Ⅱ collagen-induced arthritis (CIA) model, successful modeling rats 45 were randomly divided into three groups : model group , Tripterygium glycosides tablet group , thalidomide tablets group selected 10 only healthy rats the normal group , a total of four groups. Were gavage . Since starting the next day detection modeling arthritis index points and joint swelling degree ; After 4 weeks of rat lungs , heart, liver , kidneys, and synovial tissue expression of VEGFmRNA . Results : 1. Sequencing and electrophoretic bands confirmed the specificity of the PCR reaction , intra- CV ( coefficient of variation ) of 2.0-6.7 % and inter CV = 3.5-10.6 % , standard linear range of 6.6x10 ~ 7-6.6 x10 ~ 0 copies / ul, r> 0.99. 2. experiments showed that: after 4 weeks , Tripterygium glycosides tablet group , thalidomide tablets animals arthritis score and joint swelling were lower than the model group (P < 0.01 ) . TWP piece set in different parts of rat tissue expression of VEGF mRNA levels decreased compared with the model group were significantly different (P <0.05), and thalidomide tablets were no differences (P> 0.05). Conclusion: This study successfully established to detect VEGF in real-time PCR method, with high sensitivity, high specificity , high repeatability characteristics. TWP can be suppressed by reducing the expression of VEGF in excessive production of blood vessels and thus play a role in anti- rheumatoid arthritis .
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