Mechanism Research Of Xinfeng Capsule On Rheumatoid Arthritis By PI3K/AKT/mTOR Signaling Pathway Mediated By BAFF/BAFF-R

1 ObjectiveTo observe the changes of clinlcal symptom,laboratory index,cell factor,B cell function and PI3K/Akt/mTORC signaling pathway in RA patients'peripheral blood.Analysing the changes of B cell function and the correlation between B cell activation with others observation indicators,evaluating the clinical efficacy with XFC,estimating the mechanism with XFC acted on RA by PI3K/AKT/mTOR sinaling pathway mediated by BAFF/BAFF-R,to provide laboratory basis for the treatment of RA with traditional Chinese medicine.2 MethodsAdopt the method of clinical randomized grouping research.60 cases of RA patients which choosed from Anhui Provincial Hospital of Chinese Medicine were randomly divided into two groups,the XFC group and the LEF group,30 cases in each group,all cases are meet the criteria and classification with RA proposed by ACR/EULAR in 2010.20 cases normal person which from the MEC of Anhui Provincial Hospital of Chinese Medicine.Recording all cases datas of ages,genders and courses.Give the marks of the TCM symptoms,quality of life and the situation of anxiety and depression in RA patients by questionnaire.The levels of routine blood test were dete-cted by automatic blood analyzer.Using automatic biochemical Analyzer to detect liver and kidney function.Using electrocardiograph to detectelectrocardiogra of patients.B,B%,CD19~+CD81~+,CD19~+CD40~+were measured by flow cytometry.Using themethodsofELIAStodetectthelevelsof IL-1?,IL-2,IL-6,IL-10,BAFF,BAFF-R,PI3K,Akt,mTORC1,PDK1,PTEN,CCP-AB.Usin g automatic biochemical analyzer to detect the levels of RF,IgM,IgA,IgG,C3,C4 and CRP.Using automatic ESR analyzer to detect the level of ESR level.Calculating DAS-28 scores.3 Results3.1 The Changes of B Cell Function,laboratory index,cell factor and PI3K/AKT/mTORC signaling patnway in RA PatientsCompared with normal group the absolute number of B cells,B cell percentage,CD19~+CD81~+,CD19~+CD40~+,RF,CCP-AB,IgG,IL-1?,IL-2,IL-6,BAFF,BAFF-R,PI3K,A KT,mTORC1,PDK1 were increased obviously(P<0.05)and IL-10,PTEN were decreased obviously(P<0.01)in RA patients'peripheral blood.3.2 Relativity of B Cell Function in RA PatientsCorrelation analysis showed that in RA patients'peripheral blood the absolute number of B cell with RF,CCP-AB,ESR,IL-1?,IL-6,BAFF,BAFF-R,PI3K,mTORC1,PDK1 were positively correlated(P<0.05)and with PTEN were negatively correlated(P<0.05).The B cell percentage with IgG,CRP,IL-1?were positively correlated(P<0.05)andwithIL-10,PTENwerenegatively correlated(P<0.05).CD19~+CD81~+withC3,BAFF,AKTwerepositively correlated(P<0.05).CD19~+CD40~+with CCP-AB,IL-2,IL-10 were positively correlated(P<0.05)in 60 RA patients'peripheral blood.BAFF with PI3K were positively correlated(P<0.05)and with PTEN were negatively correlated(P<0.05).The symptom integral of arthralgia,fatigue and fatigue and deep thready pulse and total quality integral of quality of life,HAQ,HAMA,HAMD with the absolute number of B cell were positively correlated(P<0.05).Arthroncus,Joint tenderness,food intake reduction with the B cell percentage were positively correlated(P<0.05).Arthroncus,thin sloppy stool,social function integration,HAMD with CD19~+CD81~+were positively correlated(P<0.05).Nocturnal pain aggravation,the dark red tongue and greasy fur with CD19~+CD40~+were positively correlated(P<0.05).3.3 The comparison of therapeutic effect and the changes of observation index before and after treatment in two groups with RA patients3.3.1 The comparison of therapeutic effect in two groups with RA patientsAfter treatment the total effective rate,explicit efficiency and effective percentage in XFC group are 83.3%/60.0%/23.3%and in LEF group are 80.0%/36.7%/43.3%.Explicit efficiency in XFC group were increased more than in LEF group(P<0.05).Comparedwithbeforetreatmentthesymptomintegralof arthralgia,arthroncus,Jointtenderness,morningstiffness,nocturnalpain exacerbation,Joint weight,fatigue and fatigue,food intake reduction,thin sloppy stool,dark red tongue and greasy fur,the total integral of quality of life and its subitems integral(physiologic function,social function,mental function and health self-awareness ability),HAQgrade,HAMDgradeinXFCgroupweredescended obviously(P<0.05).Compared with before treatment the symptom integral of arthralgia,arthroncus,Joint tenderness,morning stiffness and Joint weightsymptom,the total integral of quality of life and its subitems integral(physiologic function and social function,mental function),HAQ grade in LEF group were descended obviously(P<0.05).ComparedwithLEFgroupthesymptomintegralof arthralgia,morning stiffness,Joint weight,food intake reduction and dark red tongue and greasy fur,he total integral of quality of life and its subitems integral(social function,mental function and health self-awareness ability,HAMD grade in XFC group were descended more(P<0.05).3.3.2 The changes of observation index before and after treatment in two groups with RA patientsCompared with before treatment the absolute number of B cell,B cell percentage,CD19~+CD81~+,CD19~+CD40~+,RF,CCP-AB,IgG,IgM,ESR,CRP,DAS-28,IL-1?,IL-2,IL-6,BAFF,BAFF-R,PI3K,AKT,mTORC1 and PDK1 in XFC group were descended obviously(P<0.05).Compared with before treatment the absolute number of Bcell,CD19~+CD81~+,CD19~+CD40~+,ESR,CRP,RF,CCP-AB,IgG,IgA,IgM,DAS-28,IL-1?,I L-2,IL-6,BAFF,BAFF-R,PI3K,AKT and PDK1 in LEF group were descended obviously(P<0.05).IL-10 in two group were inescended significantly(P<0.01).PTEN in XFC group were were inescended significantly(P<0.01).Compared with LEF group,B cell percentage,CD19~+CD40~+,CCP-AB,IgG,AKT,mTORC1and PDK1 in XFC group weredescendedmore(P<0.05).IL-10inXFCgroupwereinescended more(P<0.05).Compared with XFC group IL-2 inLEF group were descended more(P<0.05).4 Conclusion4.1 B cell level and its activation marker level,immune inflammation level in RA patients were inescended obviously,pointing out that B cell in RA patients were activated abnormally.4.2 The abnormal activated level of B cell in RA patients were related with the increased express of BAFF/BAFF-R,PI3K/Akt/mTOR,IL-1?,IL-2,IL-6 and the decreased express of IL-10.The increased integrals of TCM symptoms,AQT,HAQ,HAMAand HAMD have respect to activated B cell.4.3 The effective percentage with XFC is 83.3%and can significantly modulate clinical symptom,TCM symptoms,life quality and anxious/depression emotion in RA patients.XFC can significantly decreased the B cell level and its activation marker level.4.4 The mechanism of XFC regulate B cell function in RA patients:Acrossing the reduced expressing of PDK1 and BAFF/BAFF-R,increased expressing of PTEN,to down-regulation PI3K/Akt/mTORC which improving the levels of IL-10 and decreased the levels of IL-1,IL-2 and IL-6 possibly,then inhibited the excessive growth,proliferation and activation of B cells through the down-regulation the absolute number of B cell,B cell percentage,CD19~+CD81~+,CD19~+CD40~+.