| Objective The molecular probe 99mTc-3PRGD2 was used to observe the change of imaging agent uptake before and after the treatment of rheumatoid arthritis by Mongolian medicine Sendeng-4 decoction,and the results were compared with laboratory serological,pathological and immunohistochemical results.It is proved that 99mTc-3PRGD2 imaging is an imaging method for screening new targeted drugs for anti-synovial angiogenesis,laying theoretical foundation for Mongolian medicine clinical treatment of rheumatoid arthritis and establishing an in vivo screening and evaluation model to verify the efficacy of Mongolian medicine.Methods 1.Healthy adult female SD rats were randomly divided into two groups:collagen induced arthritis?CIA?group and blank control group.The CIA group rats model were established by using bovine type II collagen emulsion immune induction method.2.Successfully modeled CIA rats were randomly divided into a recombinant human type II tumor necrosis factor receptor-antibody fusion protein?etanercept?treatment group,a Mongolian medicine compound Sendeng-4 decoction treatment group,and a control group without any drug treatment group?hereinafter referred to as"untreated group"?.3.We observed the general state and local joint swelling degree of experimental rats,measured body weight?WT?,evaluated arthritis index?AI?,and used ROI technique to measure and analyze the changes of99mTc-3PRGD2 radioactivity count ratio?T/NT?of limb joints?T?/mediastinum?NT?before and after treatment.4.The levels of serum VEGF,TNF-?and???3 in experimental rats were detected by enzyme linked immune sorbent assay?ELISA?before and after treatment.5.After successful modeling,some rats in the CIA group,all rats in the blank control group and all rats after treatment were killed to make pathological sections of the distal joint,hematoxylin-eosin?HE?staining and synovium hyperplasia score.The expression levels of VEGF,TNF-?,???3,CD31 and CD34 in synovial tissue of experimental rats before and after treatment were detected by immunehistochemical method,and the staining results were evaluated by semi-quantitative integral method.6.SPSS21.0 statistical software was used to analyze the data.The measurement data in accordance with normal distribution are described by"x?s",and the comparison between groups is described by t-test and analysis of variance;the measurement data that do not meet normal distribution are described by M(P25,P75),and the comparison between groups is described by non-parametric test.The correlation between the two variables was analyzed by bivariate correlation analysis,bivariate normal distribution selected Pearson correlation coefficient,non-bivariate normal distribution selected Spearman correlation coefficient.The difference was statistically significant when the test level was?=0.05 and P?0.05.Results 1.Of the 176 SD rats in the CIA group?13 died?,95 were successfully established,with a modeling rate of approximately 58%.The clinical symptoms of rats in the CIA group were significant,and the scores of synovial cell proliferation and the expression levels of VEGF,???3,TNF-?,CD31 and CD34 in synovial tissue in the control group were significantly different from those in the control group?P<0.05?.2.After modeling,the levels of serology VEGF,???3 and TNF-?in the CIA group were 308.74±73.50,26.44±6.41,378.33±78.31 respectively,which were significantly higher than those in the blank control group?P<0.05?.3.Before modeling,the T/NT values of the joints in the CIA group and the blank control group were 0.226±0.031 and 0.217±0.030,respectively.There was no significant difference between the two groups?P>0.05?.Two weeks after the injection of the modeling drug,the T/NT values of the joints in the CIA group and the blank control group were 0.460±0.090,0.211±0.035,respectively;after 4weeks,the model was 0.693±0.148,0.215±0.029,respectively.The differences were statistically significant?P<0.05?.4.After 4weeks of modeling,the T/NT value of diseased joint in CIA group was positively correlated with AI value,synovial cell proliferation score,laboratory serological levels of VEGF,???3,TNF-?and the expression levels of VEGF,TNF-?,???3,CD31 and CD34 in synovial tissue?r=0.461?0.609?0.531?0.375?0.471?0.626?0.703?0.706?0.613?0.729,P<0.05?5.The T/NT in CIA group decreased after treatment with etanercept or Mongolian medicine Sendeng-4,which were 0.713±0.106 and 0.676±0.126 respectively before treatment.After 3weeks of treatment,they were 0.394±0.168 and 0.430±0.143 respectively.After 6 weeks of treatment,it was 0.238±0.099 and 0.260±0.937 respectively.There was no significant difference between the treatment group and the untreated group before treatment?P>0.05?.After 3 or 6 weeks of treatment,there were significant differences between the treatment group and the untreated group?P<0.05?,and there was no significant difference between the etanercept treatment group and the Mongolian medicine Sendeng-4 treatment group?P>0.05?.6.After treatment with etanercept or Mongolian medicine Sendeng-4,the involved joint neovascularization decreased and the symptoms gradually alleviated.AI value,pathological synovial cell proliferation score,laboratory serological levels of VEGF,TNF-?,???3 and the expression levels of VEGF,TNF-?,???3,CD31 and CD34 in synovial tissue decreased,and the T/NT value was positively correlated with the above indexes.?etanercept treatment group:r=0.422?0.632?0.549?0.500?0.522?0.472?0.457?0.558?0.492?0.697,P<0.05;Mongolian medicine Sendeng-4 treatment group:r=0.430?0.687?0.593?0.561?0.579?0.669?0.490?0.622?0.597?0.545,P<0.05;untreated group:r=0.619?0.864?0.549?0.796?0.619?0.582?0.644?0.648?0.764?0.605,P<0.05?.Conclusion?1?Mongolian medicine?represented by Sendeng-4 decoction?can delay the disease progression by inhibiting the formation of new blood vessels,and effectively improve clinical symptoms.?2?99mTc-3PRGD2 radionuclide imaging is a good evaluation model for in vivo screening and verifying the efficacy of Mongolian medicine. |
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