| ObjectiveThe present paper was that a real-time fluorescence quantitative RT-PCR detection method is developed for vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor2 (VEGFR2) mRNA in rats. By means of which the levels of VEGF of synovial tissue of joints of rats are detected during the derivation of collagen-induced arthritis (CIA) and the therapeutic procedure with added Xi jiao di huang decoction. The effect of added Xi jiao di huang decoction is discussed about the treatment of rheumatoid arthritis (RA).This is the theoretical significance of this lies with the actual meaning.Methods:SD rats are used in this experiment as rats with typeⅡCIA. Sixty inbred male DA rats were randomly divided into four groups:the noraml group, model group and added Xi jiao di huang group,Tripterygium glucosides group,15 rats each group. The back,foot, root of the tail of theⅡⅢandⅣgroup rats were multiple intradermal injection by mixed liquor of chicken collagenⅡ(CⅡ) emulsified with incomplete Freund's adjuvant (I FA), and they received booster immunization one week later. Then CIA rats were made. Normal group were multiple intradermal injection by physiological saline,and repeat the same action one week later,for comparison. Arthritis Index and joint swelling degree by observed every day.Drug were given two weeks later, initial immunizationNormal group were fed with distilled water,0.5ml/d;Model group were fed with distilled water,0.5ml/dAdded Xi jiao di huang group were fed added Xi jiao di huang 25 mg·kg-1·d1Tripterygium glucosides group were fed Tripterygium glucosides mixed 8.75 mg·kg-1·d1Forty-two days after the rats were sacrificed, Synovium of knee joint in hind limb were taken away for use.Cross-intron design primers and TaqMan probe, verify species specificity, establish real-time fluorescence detection method in the ABI7500 cycler, TAcloning of amplified fragments, sequencing and production of standard, analysis of the specificity, sensitivity and repeatability.Synovium mRNA was extraction and reverse transcription to cDNA as a template. All rats in the noraml group, model group and added Xi jiao di huang group, Tripterygium glucosides group receive a real-time fluorescence quantitative RT-PCR detection for their expressive condition of 42d VEGF, VEGFR2.Results processing and Statistical Analysis:VEGF and VEGFR2 mRNA copy number decibel divided by the corresponding GAPDH copy number, by VEGF and VEGFR2 mRNA relative expression. SPSS11.5 software was used for statistical analysis of data among the groups of the comparative analysis of variance, with P<0.05 for the difference was statistically significant.Results:Detemination of rats foot the same day-the results show that the ankle volume, rats foot of experiment group size smaller than the model group, compared with added Xi jiao di huang. There was a significant difference(P<0.05 or P<0.01).Show that added Xi jiao di huang decoction to inhibit the role of joint swelling.Biopsy showed hadn't inflammatory response in normal group; joint cavity model group disappeared, fibrous tissue filling; experiment group synovial hyperplasia, mild congestive. Arthritis model in rats showed that the model was successful, The effect of added Xi jiao di huang decoction is discussed about the treatment of rheumatoid arthritis (RA).Specimen test results were statistically significant differences between groups:F=24.711, P<0.01.Model rats synovial tissue VEGF,VEGFR2 expression were significantly higher than the normal group(P<0.05), synovial tissue of experimental rats in the expression of VEGF compared with model group.Conclusion:1. The effect of added Xi jiao di huang decoction is discussed about the treatment of VEGF VEGFR2 mRNA in rats.2. Added Xi jiao di huang decoction on the treatment of rheumatoid arthritis and acute scientific basis and application.
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