| 1 ObjectiveThe clinical study was conducted to observe the changes of Wnt/β-catenin signaling pathway,cytokines and the effects of Xinfeng Capsule(XFC)in patients with rheumatoid arthritis(RA).To analyze the correlation between Wnt/β-catenin signaling pathway and other observed indicators in RA patients,to observe the clinical efficacy of XFC in the treatment of RA,to evaluate the effect of XFC on the observed indicators,and to explore the pathogenesis of RA based on Wnt/β-catenin signaling pathway.And the mechanism of action of XFC in the treatment of RA.2 MethodsSixty patients with RA were enrolled in the Department of Rheumatology,First Affiliated Hospital of Anhui University of Traditional Chinese Medicine,from November 2017 to November 2018.Sixty patients with RA were randomly assigned to the Xinfeng capsule treatment group(XFC group)and the leflunomide control group(LEF group).The 30 patients in each group were selected to meet the 2010 American College of Rheumatology(ACR).And the latest RA classification criteria and scoring system proposed by the European Union Against Rheumatology(EULAR).At the same time,20 normal people from the physical examination center of the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine were selected as the healthy control group.The age,gender and course of the healthy person of 20 patients were recorded.Questionnaire and scale score were used for each patient.Ask in detail,record TCM symptoms and quality of life,and rate them.The indicators and methods of experimental observation are as follows: 1 Safety indicators: automatic blood analyzer is used to check blood routine;automatic biochemical analyzer is used to check liver and kidney function.2 Clinical indicators: detection of hypersensitive-C-reactive protein(hs-CRP),rheumatoid factor(RF),anti-cyclic citrullinated peptide antibody(CCP-AB),(GPI),(IgM)using an automatic biochemical analyzer Levels of(IgA),(IgG),(C3),and(C4);detection of erythrocyte sedimentation rate(ESR)using an automated erythrocyte sedimentation analyzer;calculation of DAS based on joint symptoms,ESR levels,and pain(VAS)scores-28 score.3 The levels of tumor necrosis factor-α(TNF-α),interleukin-1(IL-1)and interleukin-6(IL-6)were detected by ELISE method.Detection of β-catenin,secreted protein-1(DKK1),glycogen synthase kinase-3β(GSK-3β),secretory glycoprotein-3α(Wnt3α),and secretory glycoprotein by WB assay Expression levels of 5α(Wnt5α),bone morphogenetic protein-2(BMP-2),and matrix metalloproteinase-13(MMP-13)proteins.3 Results 3.1 Changes of laboratory indicators,Wnt/β-catenin signaling pathway,and cytokines in patients with RACompared with the NC group,the levels of RF,ESR,hs-CRP,CCP-AB,IgG,(C3),and(C4)in peripheral blood of 60 patients in the RA group were significantly higher than those in the normal group(P<0.01).Compared with the NC group,the expression levels of β-catenin,GSK3β,Wnt5α,Wnt3α,MMP-13 and BMP-2 in the peripheral blood of 60 patients in the RA group were significantly higher than those in the normal group(P<0.01),while DKK1.The expression was significantly reduced(P <0.01).Compared with the NC group,the expression levels of IL-1,IL-6 and TNF-α in peripheral blood of 60 RA patients were significantly higher than those in the normal group(P<0.01).3.2 Correlation between Wnt/β-catenin signaling pathway in RA patients 3.2.1 Correlation analysis between Wnt/β-catenin signaling pathway index and local joint symptom score in RA patientsCorrelation analysis showed that the expression level of β-catenin in RA patients was positively correlated with joint swelling,joint pain,joint tenderness,joint morning stiffness score and DAS score(P<0.05).GSK3β expression level was positively correlated with DAS score and joint pain score.(P<0.05);Wnt5α expression level was positively correlated with joint swelling and joint morning stiffness(P<0.05);Wnt3α expression level was positively correlated with DAS score and joint tenderness score(P<0.05);DKK1 expression level and DAS score,The joint morning stiffness was negatively correlated(P<0.05);BMP-2 expression was positively correlated with DAS score and joint pain score(P<0.05);MMP-13 expression level was positive with joint swelling,joint morning stiffness score and DAS score.Relevant(P<0.05).The remaining signal pathway indicators were not significantly different from the local joint symptom scores(P>0.05).3.2.2 Correlation analysis between Wnt/β-catenin signaling pathway index and systemic symptom score in RA patientsCorrelation analysis showed that the expression level of β-catenin in RA patients was positively correlated with loss of appetite and joint reintegration(P<0.05).The expression of GSK3β was positively correlated with loss of appetite,fatigue,and joint weight(P<0.05).The expression level of Wnt5α was positively correlated with the index of suffocation and post-prandial bloating(P<0.05).The expression level of Wnt3α was positively correlated with the fatigue-depletion integral(P<0.05).DKK1 was negatively correlated with stool thinning and joint re-integration.(P<0.05);BMP-2 expression level was positively correlated with postprandial bloating and joint weight accumulation(P<0.05);MMP-13 expression level was positively correlated with less gas laziness and joint weight integration(P<0.05)..There were no significant differences in the rest of the signaling pathways and systemic symptom scores(P>0.05).3.2.3 Correlation analysis between Wnt/β-catenin signaling pathway index and quality of life score in RA patientsCorrelation analysis showed that the expression level of β-catenin was positively correlated with psychological function and HAQ score in RA patients(P<0.05).The expression level of GSK3β was positively correlated with physiological function and HAQ score(P<0.05);Wnt5α expression level and social function There was a positive correlation with HAQ score(P<0.05);Wnt3α expression level was positively correlated with psychological function and social function score(P<0.05);BMP-2 expression level There was a positive correlation between social function score and healthy self-recognition ability score(P<0.05);MMP-13 expression level was positively correlated with social function score and healthy self-recognition ability score(P<0.05);DKK1 was negatively correlated with AQT and social function score(P<0.05).The remaining signal pathway indicators were not significantly different from the quality of life scores(P>0.05).3.2.4 Correlation analysis between Wnt/β-catenin signaling pathway indicators and laboratory indicators in RA patientsCorrelation analysis showed that the expression of β-catenin in RA patients was positively correlated with ESR,RF,hs-CRP and CCP(P<0.05).The expression of GSK3β was positively correlated with ESR and RF(P<0.05).The expression level of Wnt5α was correlated with hs-CRP and CCP were positively correlated(P<0.05);Wnt3α expression was positively correlated with ESR,RF and IGA(P<0.05);DKK1 expression was negatively correlated with ESR,RF and hs-CRP(P<0.05);The expression level of BMP-2 was positively correlated with ESR,hs-CRP and CCP(P<0.05).The expression level of MMP-13 was positively correlated with ESR and hs-CRP(P<0.05).There were no significant differences between the other signal pathway indicators and the laboratory indicators(P>0.05).3.2.5 Correlation analysis between Wnt/β-catenin signaling pathway markers and cytokines in RA patientsCorrelation analysis showed that the expression of β-catenin was positively correlated with IL-6 and TNF-α in RA patients(P<0.05);the expression of GSK3β was positively correlated with IL-1 and IL-6(P<0.05);Wnt5α expression The level was positively correlated with IL-6(P<0.05);the expression level of Wnt3α was positively correlated with IL-1 and TNF-α(P<0.05);the expression level of DKK1 was negatively correlated with TNF-α(P<0.05);BMP-2The expression level was positively correlated with IL-1(P<0.05);the expression level of MMP-13 was positively correlated with IL-1(P<0.05).The remaining signaling pathway indicators were not significantly different from those of cytokines(P>0.05).3.3 The mechanism of Xinfeng Capsule in the treatment of RA 3.3.1 Clinical efficacy of Xinfeng Capsule in patients with RAAfter treatment,the total effective rate of the XFC group was 90%,the marked effective rate was 63.3%,the effective rate was 26.7%,the total effective rate of the LEF group was 83.3%,the marked effective rate was 43.3%,and the effective rate was 40%.Compared with the LEF group,the total effective rate of the XFC group was equal to that of the LEF group,and the marked effective rate was higher(P < 0.05).3.3.2 Influence of Xinfeng Capsule on TCM Syndrome of RA PatientsCompared with pre-treatment,the joint pain,joint swelling,joint tenderness,morning stiffness,loss of appetite,lack of gas laziness,fatigue,post-prandial bloating,loose stools,and joint weight reduction were significantly decreased after treatment.P < 0.01).Compared with the LEF group,the XFC group had less appetite loss,less gas laziness,fatigue,and joint weight reduction(P<0.01).Joint pain,joint tenderness,and morning stiffness scores also decreased significantly(P<0.01).<0.05).3.3.3 Effect of Xinfeng Capsule on quality of life in patients with RACompared with pre-treatment,the levels of AQT,physiological function,social function,psychological function,health self-awareness and HAQ score were significantly decreased after treatment(P<0.01).Compared with the LEF group,the AQT and HAQ scores in the XFC group decreased significantly(P<0.01),and the social function scores also decreased significantly(P<0.05).3.3.4 Effect of Xinfeng Capsule on Laboratory Index of RA PatientsCompared with pre-treatment,the levels of RF,CCP-AB,GPI,C3,hs-CRP,ESR and DAS-28 were significantly decreased in the XFC group(P<0.01),and the C4 level was also decreased(P<0.05).The levels of CCP-AB,IgG,IGA,C3,hs-CRP,ESR and DAS-28 were significantly decreased in the LEF group(P<0.01),and the RF level was also decreased(P<0.05).Compared with the LEF group,the RF,CCP-AB,hs-CRP,ESR levels and DAS-28 scores in the XFC group were significantly lower(P<0.05).3.3.5 Effect of Xinfeng Capsule on the Levels of Wnt/β-catenin Signaling Pathway in RA PatientsCompared with pre-treatment,the expression levels of β-catenin,GSK-3β,Wnt3α,Wnt5α,BMP-2 and MMP-13 in XFC group decreased significantly(P<0.01),and the expression level of DKK1 increased significantly(P< The expression levels of β-catenin,GSK-3β,Wnt3α and MMP-13 in the LEF group were significantly decreased(P<0.01),and the expression level of DKK1 was significantly increased(P<0.01).Compared with LEF group,the expression levels of β-catenin,GSK-3β,Wnt5α,BMP-2 and MMP-13 in XFC group decreased more significantly(P<0.05),and the expression level of DKK1 increased more significantly(P<0.05).3.3.6 Effect of Xinfeng Capsule on the level of peripheral inflammatory factors in patients with RACompared with pre-treatment,the expression levels of IL-1,IL-6 and TNF-α were significantly decreased after treatment(P<0.01).Compared with LEF group,the expression levels of IL-6 and TNF-α in XFC group decreased more significantly(P<0.05).4 Conclusion 4.1 The expression levels of β-catenin,GSK-3β,Wnt3α,Wnt5α,BMP-2 and MMP-13 in peripheral blood of patients with RA increased significantly,and the expression level of DKK1 decreased significantly,suggesting that Wnt/β-catenin is abnormally activated,indicating that RA patients Bone destruction may be associated with abnormal activation of Wnt/β-catenin.4.2 The expression levels of pro-inflammatory cytokines IL-1,IL-6 and TNF-α in RA patients increased,RF,CCP-AB,C3,ESR and hs-CRP levels increased,DAS-28 score,TCM symptom scores,The increase in quality of life score was positively correlated with β-catenin,GSK-3β,Wnt3α,Wnt5α,BMP-2,and MMP-13,and negatively correlated with DKK1.4.3 The clinical efficacy of XFC in the treatment of RA is obvious,the efficiency is better than LEF,can effectively reduce RF,CCP-AB,C3,ESR,hs-CRP,DAS-28 score,TCM symptom score,quality of life score,can effectively reduce IL-1,IL-6 and TNF-α expression levels,inhibiting the body’s inflammatory response.4.4 The mechanism of XFC in improving the condition of RA patients may be through down-regulating the expression levels of β-catenin,GSK-3β,Wnt3α,Wnt5α,BMP-2 and MMP-13 proteins,increasing the expression level of DKK1 protein and inhibiting the Wnt/β-catenin signaling pathway.Over-activation,down-regulation of the expression levels of pro-inflammatory factors such as IL-1,IL-6 and TNF-α,inhibits the body’s autoimmune inflammatory response,inhibits the progression of bone destruction,thereby controlling the progression of RA and reducing its progression.Damage to the target organ. |
PDF Full Text Request