| Objective: To complete Freund's adjuvant(complete freund's,adjuvant,CFA)in adjuvant arthritis induced by(adjuvant arthritis,AA)rat model for experimental basis for clinical treatment of rheumatoid arthritis(rheumatoid arthritis,RA)-the effective prescription of Tonifying the kidney and clearing Luohuo blood as the experimental object,through the observation profit kidney Qingluo Huoxue Decoction with different concentrations on adjuvant arthritis(AA)rat model of morphology,inflammatory cytokines,ankle bone damage and synovial and cartilage tissue pathological damage,preliminary study on effect and mechanism of Yishen Qingluo Huoxue Decoction on inflammatory reaction in acute AA in the joint,provide reliable experimental basis for clinical treatment.Methods: the rats except the normal group right toe after intradermal injection of CFA in the rat AA.After the model was copied successfully,the rats were randomly divided into the model group,the high,middle and low dose group of Yishen Huoxue Prescription,and the Lei Gongteng group,and the corresponding drugs were given to the treatment group,28 d.Rats were observed the changes of body weight,arthritis index,swelling,X-ray observation of the ankle joint lesions in rats;after injection of blood,using ELISA method to detect serum tumor necrosis factor alpha(tumor necrosis factor alpha,TNF-alpha),interleukin 1 beta(interleukin-1 IL-1 beta),interleukin 10(interleukin-10)concentration;spleen,spleen coefficient calculation;take the ankle partto do pathological examination,a part of the Western Blot method detecting ankle TNFprotein.Results: 1 the general conditions of rats and weight changes: compared with the normal group,the rats in the model group.The spirit of Wei,yellow hair less shiny,limited activity;Yishen Huoxue Qingluo different dose group and Tripterygium group was significantly lower than that of model group in activity limitation on the appropriate relief.Compared with the normal group,the weight of each group increased slowly.Compared with the model group,the weight gain of each treatment group was improved(P<0.05);2.The arthritis index in rats of 2 groups: compared with the model group,different concentration Yishen Qingluo Huoxue Decoction group and Tripterygium group AI decreased(P<0.05);compared with Tripterygium group,Yishen Huoxue Qingluo low dose group AI was higher(P<0.05)and high dose of Yi;renal Qingluo Huoxue group AI(P>0.05)is improved.3.rats in each group were swelling comparison: compared with the model group,different concentration of Yishenqing glucoside tablets group Huoxue Decoction group and Lei Gongteng collaterals paw swelling were improved(P<0.05);Yishenqing between complexing Huoxue Decoction at different doses on the swelling of the improvement was dose-dependent,tonifying the kidney Qingluo Huoxue Decoction high dose group and the Lei Gongteng group multi glycoside tablet swelling similar improvement(P>0.05),medium and low dose group improved weak Lei Gongteng multi glycoside tablet group(P<0.05).4.rats ankle X-ray score: compared with the normal group,the rats in the model group X-ray ankle score was significantly increased(P<0.01);compared with the model group,two dose,Qingluo Yishen Huoxue Decoction group and Tripterygium group on ankle joint X-ray score decreased(P<0.01),showed that to protect the ankle of AA rats.5.rats spleen index: compared with normal group,spleen index of model group increased in AA rats(P<0.01);compared with the model group,Yishen Huoxue Qingluo three dose group and Tripterygium group spleen coefficient decreased(P<0.01);compared with Tripterygium group,kidney Qingluo huoxuefang in low and medium dose group(P<0.01),spleen coefficient was higher in high dose of Yishen Huoxue group Qingluo spleen coefficient were similar,indicating Qingluo Yishen Huoxue Recipe high dose group and Tripterygium group to reduce AA rat spleen coefficient equivalent efficacy(P>0.05).6.groups of rats serum TNF-alpha,IL-1 beta,IL-10 level: compared with the normal group,increased inflammatory cytokines TNF-,IL-1 beta,IL-10 concentration in serum of rats in model group,the difference was statistically significant(P<0.05);compared with the model group,Yishen Huoxue Qingluo three dose group and Tripterygium group of inflammatory factor IL-1 beta,TNF-alpha concentration decreased,IL-10 concentration increased,the difference was statistically significant(P<0.05);compared with Tripterygium group,Yishen Huoxue Qingluo different dose groups of serum proinflammatory cytokines,alpha IL-1 beta TNF-concentration increased,the difference was statistically significant(P<0.05);compared with Tripterygium group,Yishen Qingluo Huoxue Decoction high dose group serum anti inflammatory cytokines IL-10 concentration change was not statistically significant(P>0.05).7.rats ankle pathology score: compared with normal group,the ankle joint of rats in the model group HE AA staining revealed a large number of inflammatory cells covered,synovial membrane and marked hyperplasia of fibrous tissue,increasing blood vessels,a large number of pannus formation,cartilage erosion and bone destruction,pathological score high(P<0.05);compared with the model group,high dose of Yishen Qingluo Huoxue Decoction group and Tripterygium group obviously improved ankle injury(P<0.05),and low dose group to reduce the pathological damage is not obvious(P>0.05);high dose of Yishen Qingluo Huoxue Decoction group and Tripterygium tablets in a group of pathological injury improvement(P>0.05).8.The expression of 8 rats synovial TNF-protein: compared with normal group,the expression of TNF-protein in model group increased significantly(P<0.001);compared with the model group,Yishen Huoxue Qingluo and different concentration of Tripterygium group TNF-protein expression were decreased(P<0.01,P<0.001);and Tripterygium group,high dose of Yishen Huoxue Qingluo group TNF-expression(P>0.05),close to the middle and low dose of Yishen Huoxue Qingluo group TNFexpression was higher(P<0.05).Conclusion:1.Yishen Huoxue Decoction can increase the body weight of AA rats,reduce the arthritis index of AA rats,the degree of swelling and spleen coefficient 2.By X-ray observation of each group of rats found that high and medium dose of Yishen Huoxue Prescription on osteoporosis,bone destruction,joint space narrowing,joint swelling were significantly improved 3.Yishen Huoxue Decoction could decrease the levels of proinflammatory cytokines IL-1 and promote the level of inflammatory cytokines IL-10 by reducing the levels of pro-inflammatory cytokines TNF-alpha,so as to regulate the balance between the destructive and protective cytokines 4.The pathological changes of the ankle joint were observed by optical microscope.The pathological damage of the ankle joint in the high and medium dose group was significantly improved 5.Yishen Huoxue Qingluo by expression of TNF-protein inhibited synovial tissue,improve the synovial inflammation,reduce pannus during acute period of AA generation and play an important role in inhibiting bone destruction. |
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