| Objective:In this study,the effects of Sanbi Decoction on foot volume,arthritis index score,ankle joint pathological morphology and STAT3/RORγt signaling pathway related factors in rheumatoid arthritis model rats were observed to explore the mechanism of Sanbi Decoction in the treatment of rheumatoid arthritis.Methods:Rheumatoid arthritis model rats were prepared with Freund ’s complete adjuvant.Sixty rats were randomly divided into 6 groups : blank group,model group,low-dose Sanbi Decoction group(1.83 g/kg·d),medium-dose Sanbi Decoction group(3.67 g/kg·d),high-dose Sanbi Decoction group(7.35g/kg·d),and tripterygium glycosides group(10 g/kg·d).The foot volume of rats in each group was measured by self-made foot volume measuring instrument.The arthritis index score of rats in each group was evaluated.The pathological changes of ankle joint in each group were observed by HE staining.The contents of IL-6 and IFN-γ in serum of rats in each group were detected by ELISA.The expression levels of STAT3,RORγt and Foxp3 in ankle synovial tissue of rats in each group were detected by immunohistochemistry.The expression level of STAT3 m RNA in ankle synovial tissue of rats in each group was detected by RT-PCR.Results:1.Changes in foot volume of rats in each group : Compared with the blank group,the foot volume of rats in the model group,Sanbi Decoction high-dose group,Sanbi Decoction middle-dose group,Sanbi Decoction low-dose group and tripterygium glycosides group increased significantly on the 7th day after the first inflammation(P<0.05);compared with the model group,the foot volume of rats in the high-dose Sanbi Decoction group,the middle-dose Sanbi Decoction group and the tripterygium glycosides group decreased significantly on the 14 th,21 st,28 th and 35 th day after the initial inflammation(P<0.05),and the foot volume of rats in the low-dose Sanbi Decoction group decreased significantly on the 21 st,28 th and 35 th day after the initial inflammation(P<0.05).Compared with the low-dose group of Sanbi Decoction,the foot volume of rats in the high-dose group of Sanbi Decoction,the middle-dose group of Sanbi Decoction and the tripterygium glycosides group decreased significantly on the 14 th,21 st,28 th and 35 th days after the initial inflammation(P<0.05);compared with the tripterygium glycosides group,the foot volume of the rats in the high-dose Sanbi Decoction group decreased significantly on the 21 st day after the first inflammation(P<0.05),and the foot volume of the rats in the middle-dose Sanbi Decoction group decreased significantly on the 14 th day after the first inflammation(P<0.05).2.Arthritis index score of rats in each group : Compared with the blank group,the arthritis index score of rats in the model group,Sanbi Decoction high-dose group,Sanbi Decoction middle-dose group,Sanbi Decoction low-dose group and tripterygium glycosides group increased significantly on the 7th day after the first inflammation(P<0.05),and the arthritis index score≥ 6 points,suggesting that the rheumatoid arthritis model rats were successfully induced;compared with the model group,the arthritis index scores of rats in the high-dose Sanbi Decoction group,the medium-dose Sanbi Decoction group,the low-dose Sanbi Decoction group and the tripterygium glycosides group were significantly decreased on the 14 th,21 st,28 th and 35 th days after the initial inflammation(P<0.05).Compared with the low-dose group of Sanbi Decoction,the arthritis index scores of rats in the high-dose group of Sanbi Decoction,the middle-dose group of Sanbi Decoction and the tripterygium glycosides group were significantly lower on the 14 th,21 st,28 th and 35 th days after the initial inflammation(P<0.05);compared with the tripterygium glycosides group,the arthritis index score of the rats in the high-dose group of Sanbi Decoction was significantly lower on the 21 st,28 th and 35 th days after the initial inflammation(P<0.05),and there was no significant difference in the middle-dose group of Sanbi Decoction on the 14 th,21 st,28 th and 35 th days after the initial inflammation.3.Pathological changes of ankle joint tissue in each group :In the blank group,the synovial cells were arranged neatly,the blood vessels were not dilated,the gap was not abnormal,the synovial cells were not proliferated,and there was no inflammatory cell infiltration.In the model group,the synovial layer cells were significantly increased,the arrangement was disordered,the tissue structure was destroyed,the hyperplasia was serious,the capillaries were dilated,the severe pannus was formed,and a large number of inflammatory cells were infiltrated.In the low-dose group,the arrangement of synovial cells was disordered,the thickening of synovial layer was reduced,capillaries were dilated,moderate pannus was formed,and a large number of inflammatory cells were infiltrated.The rats in the medium-dose group had mild synovial tissue hyperplasia,a small amount of pannus,and a small amount of inflammatory cell infiltration;the rats in the high-dose group had mild synovial tissue hyperplasia and a small amount of inflammatory cell infiltration;tripterygium glycosides group rats synovial cell thickening,telangiectasia,a small amount of inflammatory cell infiltration.4.Changes of IL-6 and IFN-γ in serum of rats in each group : Compared with the blank group,the contents of IL-6 and IFN-γ in serum of rats in the model group were significantly increased(P<0.05).Compared with the model group,the contents of IL-6 and IFN-γ in the serum of rats in the high-dose Sanbi Decoction group,the middle-dose Sanbi Decoction group,the low-dose Sanbi Decoction group and the tripterygium glycosides group were significantly decreased(P<0.05).Compared with the low-dose group of Sanbi Decoction,the serum levels of IL-6 and IFN-γ in the high-dose group of Sanbi Decoction,the middle-dose group of Sanbi Decoction and the tripterygium glycosides group were significantly decreased(P<0.05);compared with the tripterygium glycosides group,the contents of IL-6 and IFN-γ in the serum of rats in the high dose group of Sanbi Decoction were significantly decreased(P<0.05),and there was no significant difference in the content of serum in the middle dose group of Sanbi Decoction.5.The expression of RORγt,STAT3 and Foxp3 in synovial tissue of rats in each group : Compared with the blank group,the expression levels of RORγt and STAT3 in synovial tissue of rats in the model group were significantly increased(P<0.05),and the expression level of Foxp3 was significantly decreased(P<0.05).Compared with the model group,the expression levels of RORγt and STAT3 in synovial tissue of rats in the high-dose Sanbi Decoction group,the middle-dose Sanbi Decoction group,the low-dose Sanbi Decoction group and the tripterygium glycosides group were significantly decreased(P<0.05),and the expression level of Foxp3 was significantly increased(P<0.05).Compared with the low-dose Sanbi Decoction group,the expression levels of RORγt and STAT3 in the synovial tissue of rats in the high-dose Sanbi Decoction group,the middle-dose Sanbi Decoction group and the tripterygium glycosides group were significantly decreased(P<0.05),and the expression level of Foxp3 was significantly increased(P<0.05);compared with the tripterygium glycosides group,the expression levels of RORγt and STAT3 in the synovial tissue of rats in the high-dose Sanbi Decoction group were significantly decreased(P<0.05),and the expression level of Foxp3 was significantly increased(P<0.05).There was no significant difference in the expression level of synovial tissue in the middle-dose Sanbi Decoction group.6.The expression of STAT3 m RNA in synovial tissue of rats in each group :Compared with the blank group,the expression of STAT3 m RNA in the synovial tissue of the model group was significantly increased(P<0.05).Compared with the model group,the expression of STAT3 m RNA in the synovial tissue of rats in the Sanbi Decoction high-dose group,Sanbi Decoction middle-dose group,Sanbi Decoction low-dose group and tripterygium glycosides group was significantly decreased(P<0.05);compared with the low dose group of Sanbi Decoction,the expression of STAT3 m RNA in the synovial tissue of rats in the high dose group of Sanbi Decoction,the middle dose group of Sanbi Decoction and the tripterygium glycosides group was significantly decreased(P<0.05);compared with tripterygium glycosides group,the expression of STAT3 m RNA in synovial tissue of rats in Sanbi Decoction high dose group was significantly decreased(P<0.05),and there was no significant difference in the expression of STAT3 m RNA in synovial tissue of rats in Sanbi Decoction middle dose group.Conclusions:1.Sanbi Decoction can reduce the inflammatory cell infiltration of ankle joint in rheumatoid arthritis model rats,improve the pathological damage of synovial tissue,and reduce the foot volume and arthritis index score.2.Sanbi Decoction may regulate the expression levels of RORγt and Foxp3 by inhibiting STAT3 signaling pathway,thereby reducing the content of IL-6 and IFN-γ and improving synovial injury. |
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