| Rheumatoid arthritis(RA)is a common autoimmunity disease characterized by chronic,progressive synovitis and destructive arthropathy,the disability rate of which is so high as to harm people's life and work seriously.The etiology and pathogenesis of RA are still unclear.And there is no effective treatment or drugs for it.Paeoniae,Glycyrrhizae and Aconiti Decoction(PGAD)was firstly recorded in Treatise on Febrile Diseases.It is composed of White Peony Root,Prepared Licorice and Prepared Aconite Root,with the functions of reinforcing deficiency and reducing excess,warming and activating meridian,nurturing blood and extending sinew,relieving spasm and pain.In the clinical treatment of RA,PGAD and its modification have shown good efficacy,but the efficacy and action mechanism of PGAD are short of systematic research.In this study,we observed the therapeutic effect of PGAD on RA and investigated its mechanism of action by collagen-induced arthritis(CIA)rat model,nonspecific inflammatory model and nonspecific pain model,to provide a reference for PGAD's further clinical application.1 Pharmacodynamic study of Paeoniae,Glycyrrhizae and Aconiti Decoction on collagen-induced arthritis in ratsObjective:To observe the effects of PGAD on the foot swelling,the content of RF,CRP,IFN-?,IL-1 7,IL-4 and IL-10 in serum,the indexes of spleen and thymus and the pathological injury of the ankle joint in the CIA rat model.Methods:The CIA model in rats was induced by intradermal injection of bovine type ?collagen and incomplete Freund's adjuvant at the root of the tail.The CIA rats were randomly divided into model group,PGAD high-dose group(4.2 g/kg),PGAD medium-dose group(2.1 g/kg),PGAD low-dose group(1.05 g/kg),Tripterygium glycosides tablet group(9 mg/kg),and set the same number of rats in the normal group.From 17d after primary immunization,each group was gavaged with the corresponding drug,once a day for 28 consecutive days.Foot swelling rate and arthritis index were used to evaluate the degree of joint damage of rats.The content of RF and CRP in serum after the last administration were detected by ELISA.The dynamic changes of IFN-?,IL-1 7,IL-4 and IL-10 in the 24 days(d 24),the 31 days(d 31),the 38 days(d 38)and the 45 days(d 45)after primary immunization were detected by ELISA.After the last administration,the indexes of spleen and thymus were measured.The pathological injury of the ankle joint was examined by HE staining.Results:? Foot swelling rate and arthritis index Compared with the normal group,the foot swelling rate and arthritis index in the model group significantly increased(P<0.01).In the treatment groups with PGAD,the foot swelling rate and arthritis index significantly decreased when compared with those in the model group(P<0.05 or P<0.01).? The levels of RF and CRP Compared with the normal group,the contents of serum RF and CRP in the model group increased significantly(P<0.05).Compared with the model group,the content of RF in PGAD low-dose group and the content of CRP in PGAD high-dose group decreased(P<0.05).? The levels of IFN-?,IL-17,IL-4 and IL-10 The levels of IFN-? and IL-17 in the model group were significantly higher than those in the normal group on d 24,d 31,d 38 and d 45(P<0.05 or P<0.01).The levels of IL-4 on d 38,d 45 and the levels of IL-10 on d 24,d 38 and d 45 in the model group were higher than those in the normal group(P<0.05 or P<0.01).Compared with the model group,the levels of IFN-? in PGAD high-dose group on d 31 and d 45,in the medium-dose group on d 38 and d 45 and in the low-dose group on d 45 decreased significantly(P<0.05 or P<0.01).The levels of IL-17 in PGAD high-dose group on d 31 and d 38,in the medium-dose group on d 38 and d 45 and in the low-dose group on d 31,d 38 and d 45 were lower than those in the model group(P<0.05 or P<0.01).On d 31 and d 38,the levels of IL-4 in all dose groups of PGAD decreased significantly(P<0.05 or P<0.01).The levels of IL-10 in PGAD high-and low-dose groups on d 31,d 38 and d 45,and in the medium-dose group on d 38 and d 45 were lower than those in the model group(P<0.05 or P<0.01).? The indexes of spleen and thymus The indexes of spleen and thymus in the model group were higher than those in the normal group(P<0.05).Compared with the model group,the index of spleen of rats in PGAD high-dose group and the indexes of thymus of rats in PGAD high-and medium-dose groups decreased(P<0.05).? Pathological observation of the ankle joint Compared with the normal group,pathological changes of the ankle joint in the model group showed severe synovial inflammation,angiogenesis and cartilage destruction.Each dosage group of PGAD could relieve the pathological changes of the ankle joint and decrease the pathologic scores significantly compared with the model group(P<0.05 or P<0.01).Conclusion:PGAD can significantly reduce foot swelling,arthritis index,serum CRP and RF levels and the indexes of spleen and thymus in CIA rats.It can adjust the levels of serum IFN-?,IL-17,IL-4 and IL-10.It also can relieve the severity of synovial inflammation,angiogenesis and cartilage destruction,and decrease the pathologic score.2 Effect of Paeoniae,Glycyrrhizae and Aconiti Decoction on the MAPK signaling pathway on CIA ratsObjective:To observe the effect of PGAD on the MAPK signaling pathway in rats with collagen-induced arthritis,and to explore the mechanism of PGAD in the treatment of RA.Methods:Based on the CIA model in rats,the expressions of p38 and p-p38 in synovial tissues were detected by immunohistochemistry,and the expressions of JNK,p-JNK,ERK and p-ERK in synovial tissues were detected by Western Blot.Results:Compared with the normal group,the integral optical density(IOD)of p38 and p-p38 in the model group significantly increased(P<0.01).Compared with the model group,the IOD of p38 and p-p38 protein in all dosage groups of PGAD significantly decreased(P<0.05).Compared with the normal group,the expressions of JNK,ERK,p-ERK protein in the model group significantly increased(P<0.05 or P<0.01),the expression of p-JNK showed an increasing tendency.Compared with the model group,the expression of JNK protein in the synovial tissue of rats in PGAD medium-and low-dose groups significantly decreased(P<0.05),and the expression of p-ERK protein in all dose groups of PGAD significantly decreased(P<0.01).The expression of p-JNK and ERK protein tended to decrease.Conclusion:PGAD can down-regulate the phosphorylation level of p38 and ERK protein,thus regulate the MAPK signaling pathway in the synovial tissue.3 Anti-inflammatory and analgesic effects of Paeoniae,Glycyrrhizae and Aconiti DecoctionObjective:To research the anti-inflammatory and analgesic effects of PGAD.Methods:The anti-inflammatory effect was observed by the methods of xylene-induced ear-swelling and increasing of capillary permeability induced by acetic acid in mice.The acetic acid writhing test and hot plate test in mice were adopted to observe the analgesic effect.Results:In all dose groups of PGAD,the swelling degrees of ear induced by xylene in mice decreased significantly when compared with that in the model group(P<0.01).All dosages of PGAD could significantly inhibit the acetic acid-induced increasing capillary permeability in the abdominal cavity in mice(P<0.05 or P<0.01).All dosages of PGAD could reduce the frequency of acetic acid-induced writhing in mice(P<0.01).And the medium-and high-dose of PGAD could significantly prolong the latent period of writhing in mice(P<0.01).The hot plate pain thresholds in PGAD medium-and high-dose groups were higher than that in the model group(P<0.05 or P<0.01).Conclusion:PGAD shows notable anti-inflammatory and analgesic effects.Above results suggest that Paeoniae,Glycyrrhizae and Aconiti Decoction can reduce foot swelling,serum CRP and RF levels,and the indexes of spleen and thymus in CIA rats.It also can relieve the pathological changes of ankle joint,and shows notable anti-inflammatory and analgesic effects.Its mechanism may be related to the regulation of cytokine levels and the MAPK signaling pathway. |
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