| 1 Objective In this paper,a large number of clinical data mining,analysis,discussion of traditional Chinese medicine treatment of RA medication rules,characteristics and advantages.Through the study of RA inhibitors in patients with Th17/Treg related cytokines and cytokine signaling(SOCS1),signal transduction activator of transcription factor 3(STAT3),changes and correlation analysis of small non encoding RNA326 and syndromes of traditional Chinese medicine;to explore the mechanism of traditional Chinese medicine Xinfeng capsule orally combined with external application of Furong Paste regulation of RA in patients with Th17/Treg.2 Methods 2.1 Study on clinical data mining of RA patients with internal and external treatment of Chinese Medicine From June 2012 to October 2015 of Rheumatology,the First Affiliated Hospital of Anhui College of traditional Chinese medicine in 1080 cases of active RA patients,divided into the internal treatment of TCM group(186 cases)and combined internal and external treatment group(894 cases).The core prescription of Chinese medicine was drawn by complex network technology.The data mining technology was used to analyze the frequency,frequency,sexual flavor and tropism of drugs.Clementine 12 software Aprior module was used to analyze the correlation between the decline of drugs and laboratory indexes.Analysis of the two groups before and after treatment of high sensitivity C reactive protein by SPSS17.0 software(hs-CRP),erythrocyte sedimentation rate(ESR),alpha 1-acid glycoprotein(alpha 1-AGP),rheumatoid factor(RF)and anti cyclic citrullinated peptide antibody(CCP),immunoglobulin G(Ig G),immunoglobulin A(Ig A)and complement C3 complement C4 changes.2.2 Changes of Th17/Treg and its correlation with mi R326 in patients with RA From December 2015 to October 2016,40 patients with RA in the First Affiliated Hospital of Anhui University of traditional Chinese medicine(TCM)were enrolled in the Department of Rheumatology,and 20 healthy normal subjects from our hospital health examination center.Using enzyme-linked immunosorbent assay(ELISA)detection of serum interleukin 10(IL-10),IL-2,IL-17,transforming growth factor beta(TGF-beta),Est1,suppressor of cytokine signaling 1(SOCS1)and retinoid related orphan receptor gamma(Ro R?t)antibody,signal transduction transcription the activation of factor 3(STAT3),fork type transcription protein(Foxp3)expression of ESR detection;Westergren s method;automatic biochemical detection of hs-CRP,RF,Ig G,Ig A,C3,C4;by flow cytometry(FCM)detection of Treg cell surface molecules(CD4,CD25,Foxp3)and Th17 cell surface molecules(CD3,CD8,IL-17)expression;Western blot technique(WB)to detect the expression of Ets1,SOCS1,Ro R?t,STAT3;expression by fluorescence quantitative PCR detection of Foxp3 m RNA,IL-17 m RNA,mi R326.SPSS17.0 statistical software was used to analyze the difference and correlation between the above indexes of normal people and RA patients.2.3 Study on the mechanism of regulating Th17/Treg in patients with RA based on SOCS1/STAT3/mi R326 and Xinfeng Capsule 40 cases of RA patients were randomly divided into treatment group(Xinfeng Capsule + Furong Ointment)and control group(LEF),20 cases in each group,using a questionnaire to observe the joint symptoms,TCM symptoms and quality of life in patients with RA and HAQ integral;analysis of two groups before and after RA treatment in patients with immune inflammation index by SPSS17.0 statistical software(ESR,hs-CRP,RF,CCP-AB,a1-AGP,Ig G,Ig A),serum cytokines(IL-10,IL-2,IL-17,TGF-,beta)transcription factors(Ets1,SOCS1,Ro R?t,,STAT3)fork head protein(Foxp3)and Treg cells(CD4,CD25,sub branch Foxp3),Th17 cell surface molecules(CD3,CD8,IL-17),Foxp3 m RNA,IL-17 m RNA,mi R326 changes.3 Results 3.1 Study on clinical data mining of RA patients with internal and external treatment of Chinese Medicine 3.1.1 Traditional Chinese medicine A total of 1080 RA patients involving 300 aftertaste Chinese medicine,analysis of complex network based on the core prescription drug clinical treatment of damp heat Bizu RA patients for coix seed,tuckahoe,yam,dandelion,Herba Siegesbeckiae,Salvia miltiorrhiza,peach kernel,safflower,spatholobus stem,clematis,liquorice,Rhizoma alismatis,malt,tangerine peel,mainly divided into jianpihuashi medicine,qingrelishi drugs,Huoxue Tongluo medicine,Qufengchushi drug class four.The highest frequency of use is jianpihuashi medicine,more to the spleen and stomach,lung,taste more sweet light of peace.3.1.2 The spleen of TCM medicine and immune inflammation index falling correlation The frequency of Poria cocos,coix seed and tangerine peel was higher in Jianpi Jianpi decoction.The confidence levels of Poria and ESR and hs-CRP decreased by 70.31% and 76.21% respectively.The confidence levels of Coix seed and a1-AGP and RF decreased by 92.70% and 86.13% respectively.The confidence levels of Pericarpium Citri,C4 and Ig A were higher than 50%.In conclusion,there is a clear relationship between the index of spleen and dampness and the decrease of immune and inflammatory indexes.3.1.3 Chitin and immune inflammation index falling outside the relationship The confidence levels of hs-CRP,ESR,RF and a1-AGP decreased with 72.58%,60.77%,62.33% and 68.23% respectively.Furong Ointment was significantly associated with the decrease of inflammatory markers hs-CRP,ESR,RF and a1-AGP.3.1.4 Analysis of inflammatory and immune indexes before and after treatment in two groups Compared with before treatment,the levels of ESR,hs-CRP,1-AGP,RF,CCP-AB,Ig G and PLT in the two groups were significantly lower(P<0.05).Compared with the within group,internal and external treatment group after treatment,ESR,hs-CRP,1-AGP,RF,alpha CCP-AB,Ig G,Ig A,PLT decreased significantly,the difference was statistically significant(P<0.05).In conclusion,compared with the traditional Chinese medicine treatment,the inflammatory indexes and immune indexes decreased significantly in the internal and external treatment groups.3.2 Changes and correlation analysis of Th17/Treg and mi R326 in patients with RA 3.2.1 RA patients of Th17/Treg related laboratory indexes and the change of the serum levels of cytokines and correlation analysis Compared with the normal group,RA patients with Th17 related IL-17,IL-2,TGF-? increased significantly,Treg IL-10 and Foxp3 decreased significantly.CRP,IL-17 and Ig A were positively correlated;there was negative correlation between Foxp3 and ESR;IL-10 was negatively correlated with RF and CRP;IL-2 was positively correlated with a1-AGP and negatively correlated with Ig A;TGF-? was negatively related to beta and RF(P<0.05).Compared with the normal group,Th17/Treg in patients with RA increased significantly,and Th17/Treg was positively related to RF and a1-AGP(P<0.05).3.2.2 Surface of Th17/Treg molecular changes in RA patients and the correlation analysis and laboratory indexes Treg cell surface molecules were CD4,CD25,Foxp3,and Th17 cell surface molecules were CD3,CD8,IL-17.Compared with the normal group,RA and CD4+CD25+Treg in patients with CD4+CD25+Foxp3+Treg decreased significantly,while CD3+CD8+IL-17 +Th17 cells increased significantly(P<0.05);and CD4+CD25+Treg was negatively correlated with CCP-AB,ESR,CD4+CD25+Foxp3+Treg were negatively correlated with RF,CRP,Ig G,CD3+CD8+Th17 and ESR,CCP-AB was positively related to CD8+IL-17+Th17 and CCP-AB,Ig G positive correlation.Compared with the normal group,the CD3+CD8+IL-17+Th17/CD4+CD25+Foxp3+Treg of patients with RA increased significantly(P<0.01).3.2.3 RA patients of Th17/Treg related transcription factors of the change and correlation analysis Compared with the normal group,the Th17 related STAT3,ROR?t in patients with RA were significantly increased.STAT3,ROR?t were negatively correlated with Treg,but ROR?t was positively correlated with Th17/Treg(P<0.05).Compared with the normal group,the Treg related Ets1 and SOCS1 in patients with RA were significantly lower.Ets1 was negatively correlated with Th17,and SOCS1 was negatively correlated with Th17/Treg(P<0.05).3.2.4 Foxp3 m RNA RA patients,the change of IL-17 m RNA Compared with the normal group,RA decreased and IL-17 m RNA increased in patients with Foxp3 m RNA,the difference was statistically significant(P <0.05).3.2.5 RA patients with peripheral blood mi R326 and correlation analysis of transcription factors Compared with normal group,mi RNA326 in patients with RA was relatively high.Mi R326 was positively correlated with Th17 and SOCS1,and negatively correlated with Ets1(P<0.05).3.3 Study on the mechanism of regulating Th17/Treg in patients with RA based on SOCS1/STAT3 /mi R326 3.3.1 Two groups of RA patients before and after treatment of joint symptoms and changes of traditional Chinese medicine Compared with before treatment,the treatment group and the control group after treatment of joint symptoms,TCM syndrome score decreased(P<0.05).Compared with the control group,the treatment group after the treatment of joint pain,joint pain,morning stiffness,less gas lazy words,anorexia were significantly decreased(P<0.05).3.3.2 Two groups of RA patients' quality of life the change of the integral Compared with before treatment,the physiological function,social function,psychological function and health self cognition ability of the treatment group and the control group were decreased(P<0.05).Compared with the control group,the psychological function,health self cognition ability of the treatment group decreased(P<0.05).3.3.3 Changes of inflammatory and immune indexes in two groups of patients with RA Compared with before treatment,the levels of ESR,hs-CRP,RF,CCP-AB,a1-AGP and Ig G in the treatment group and the control group were significantly decreased(P<0.05). Compared with the control group,the ESR,hs-CRP and a1-AGP in the treatment group were significantly lower(P<0.05).3.3.4 Two groups of patients with RA of Th17/Treg related cytokines,the change of transcription factors Compared with before treatment,after treatment,Th17/Treg,IL-17,IL-2,TGF-? significantly decreased,while IL-10 increased significantly after treatment;IL-17,IL-2,TGF-? significantly reduced control(P<0.05 or P<0.01),and Th17/Treg(P>0.05,IL-10)decreased slightly increased slightly(P>0.05).Compared with the control group,the Th17/Treg in the treatment group decreased significantly(P<0.05).3.3.5 Two groups of Th17/Treg surface molecular changes in RA patients Compared with before treatment,CD4+CD25+Foxp3+Treg increased significantly after treatment(P<0.01),and CD3+CD8+IL-17+Th17 decreased significantly(P<0.05);the control group after treatment,CD3+CD8+IL-17+Th17 was slightly decreased(P>0.05),CD4+CD25+Foxp3+Treg(P>0.05)increased slightly.Compared with the control group,the CD4+CD25+T in the treatment group increased significantly(P<0.01).3.3.6 Two groups of patients with RA of Th17/Treg related expression of transcription factors Compared with before treatment,after treatment of Th17 related STAT3,ROR?t decreased significantly(P<0.05),Treg Ets1 and SOCS1 increased significantly(P<0.05);group STAT3,ROR?t was significantly decreased in control(P<0.05).Compared with the control group,the Treg related SOCS1 in the treatment group was increased significantly(P<0.05).3.3.7 Two groups of patients with RA Foxp3 m RNA,IL-17 m RNA and mi R326 changes Compared with before treatment,Foxp3 m RNA increased in treatment group,but decreased in IL-17 m RNA and mi R326(P<0.05),but in control group Foxp3 m RNA increased slightly(P>0.05).Compared with the control group,the treatment group after treatment Foxp3 m RNA increased significantly,IL-17 m RNA and mi R326 were significantly lower(P<0.05).4 Conclusion 4.1 Our hospital treatment of active RA patients with Jianpi Qingre Tongluo Qushi,use of jianpihuashi medicine,medicine,medicine qingrelishi Qufengchushi,Huoxue Tongluo drug class four drugs.Traditional Chinese medicine for invigorating spleen and dredging collaterals and oral application of Furong ointment were associated with inflammation and immune index.4.2 The activity of Th17/Treg in patients with SOCS1 increased during the active period,which showed the imbalance of Th17/Treg related cytokines,transcription factors STAT3,ROR?t,Ets1 and RA.Th17/Treg was positively correlated with disease activity index and STAT3,and negatively correlated with SOCS1.Mi R326 was positively correlated with Th17.4.3 Xinfeng Capsule Combined with Furong Ointment obviously improved the clinical symptoms and inflammatory indexes of RA patients,and the curative effect was better than that of the control group.4.4 Xinfeng Capsule Combined with Furong Ointment can obviously improve the RA in patients with Th17/Treg,the mechanism is as follows:(1)By regulating transcription factor STAT3 and ROR?t,the expression of Ets1 and SOCS1 was up-regulated,and thus regulate Th17/Treg;(2)By inhibiting the expression of Th17 molecules on the surface of IL-17,it can promote the expression of Treg molecules on the surface of Foxp3,and thus regulate Th17/Treg;(3)Regulation of mi R326 by down regulating the expression of Th17/Treg;(4)By inhibiting the expression of proinflammatory factors,promoting the expression of anti-inflammatory factors,thereby regulating Th17/Treg. |
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