Study Of Effects And Mechanism Of Xinfeng Capsule On Immunoglobulin Of Osteoarthritis Based On PI3K/Akt-mTOR And Beclin-1 Pathway

1 Objective Observation of impact of Xin Feng capsule on immunoglobulin(Ig) autophagy changes of knee osteoarthritis(KOA) from both clinical and experimental, and explore the mechanism how XFC lowering immunoglobulin of knee osteoarthritis patients based on PI3 K / Akt-m TOR pathway and Beclin-1.2 Methods2.1 Clinical Research2.1.1 Observation of 60 cases of KOA patients’ age, sex, disease duration, joint and systemic symptom scores, quality of life, mental health integration value, disease activity score, immunoglobulin and other laboratory indicators.2.1.2 Detect immunoglobulin subtypes of KOA group(60 cases) and normal control group(NC, 20 cases) using ELLSA.Detect the expression of BAFF-R in peripheral blood of KOA patients and NC with Flow cytometry(FCM).And determination of the relevant laboratory parameters in patients with KOA(Ig G, Ig A, Ig M, Hs-CRP, ESR, etc.),the integral value of symptoms and signs were observed in patients with KOA(joints,body, etc.), quality of life, health index(HAQ),, anxiety(SAS), depression(SDS)and other integral values. Using SPSS17.0 statistical software to analyze the relationship between immunoglobulin subtypes between these indicators.2.1.3 The 60 cases of hospitalized patients were randomly divided into experimental group(Xinfeng capsule group,30 cases) and control group(glucosamine group,30 cases) were observed before and after treatment of immunoglobulin subtypes Ig G1, Ig G2, Ig G3,Ig G4, BAFF / BAFF-R, joint, systemic symptom score values, quality of life integral value, HAQ and SAS, SDS integral value and improvement of KOA related laboratory parameters(Ig G, Ig A, Ig M, CRP, ESR etc.).2.2 Experimental Study2.2.1 The 40 rats were randomly divided into four groups: normal control group(NC),model group(MC), Xinfeng Capsule group(XFC), glucosamine group(GS), n = 10.In addition to the normal control group, inject 20 μL 4% papain solution with 0.03 mol / L L- cysteine solution mixed solution to the right of each animal knee articular, modeling into the KOA models. 19 days after the modeling, each treatment group began to be administered for consecutive 30 days.2.2.2 Observe spontaneous activity of rats in each group status and behavior, body weight, immunoglobulin subtypes, cytokines, cartilage, thymus, spleen autophagy gene and protein expression and the impact of Xinfeng Capsule on them.3 Results3.1 Clinical studies3.1.1Changes and correlation of immunoglobulin in the KOA patient Compared with 20 normal controls,Ig G1, Ig G2, Ig G3 levels increased in KOA patients(P <0.01 or P <0.05), Ig G4 levels elevated, but not statistically significant, BAFF-R and BAFF levels were significantly increased(P <0.01 or P <0.05).Correlation analysis shows,immunoglobulin subtype and disease duration, age was no significant correlation; Ig G1, Ig G3, Ig G4 and quantify symptoms integral positive correlation(P <0.05 or P <0.01), Ig G1, Ig G4 was positively correlated with Lequesne MG(P <0.05);Ig G1 and physiological functions(RP),role emotional(RE),social functioning(SF)was negatively correlated(P <0.05), Ig G2 and general health(GH), energy(VT)negative correlation(P <0.05),Ig G3 is no significant correlation between the dimensions,Ig G4 and emotional function(RE), bodily pain(BP) was positively correlated(P <0.05);BAFF-R and Lequesne MG,bodily pain,Ig G3,ESR,Hs-CRP and other positive correlation(P <0.05 or P <0.01), and general health(GH), physiological function(PF),role physical(RP),emotional Functions(RE),social functioning(SF),mental health(MH) and other indicators were negatively correlated(P<0.05 or P<0.01); BAFF and bodily pain(BP), SDS, Ig G2 was positively correlated with general health(GH),physiological function(PF),role physical(RP),role emotional(RE),social functioning(SF), mental health(MH) and other indicators were negatively correlated(P <0.05 or P<0.01);Ig G1 is positive correlate with Ig A, Ig M(P <0.05), Ig G2 and Hs-CRP, Ig M positive correlation(P <0.01 or P <0.05), Ig G3 showed no significant correlation, Ig G4 and Ig M positive correlation(P <0.01).3.1.2 Clinical efficacy of Xinfeng capsuleon KOA After treatment, the experimental group and the control group, the total efficiency of93.3%, 76.7% respectively, no significant difference(P> 0.05) in the total effect; the experimental group were efficiency(60.0%) was significantly higher(36.7%)(P <0.05).3.1.3 Effect of Xinfeng Capsule on immunoglobulin in KOA patients.Before treatment, the two groups of patients KOA immunoglobulin subtype was no significant difference(P> 0.05). After treatment, the immunoglobulin subtypes were decreased. Compared with before treatment, GS group can significantly improve Ig G3;XFC can significantly decrease Ig G1, Ig G3, and in terms of reducing the Ig G1, XFC group is better than GS group(P <0.05).3.1.4 The impact of Xinfeng Capsule on joint and systemic symptom score values of KOA patients Compared with the pre-treatment, both groups improved significantly after treatment with systemic symptoms, malaise, fatigue, less gas lazy words, the joint weight, loss of appetite, abdominal distention after eating, loose stool score was significantly lower( P<0.01). Compared with the control group after treatment, the experimental group after treatment, loss of appetite, less gas lazy words, some indicators, such as the degreeof integration bloating after eating more obvious improvement(P <0.01).3.1.5 The impact of Xinfeng Capsule on quality of life and mental health status of KOA patients.Compared with pre-treatment, physical function, mental function, self-awareness capability, HAQ, SAS, SDS integration were significantly lower(P <0.05 or P <0.01).After treatment compared with the control group, the experimental group after treatment of physiological function, social function, quality of life, SDS points and other decreased(P <0.05 or P <0.01).3.1.6 Impact of Xinfeng Capsule on BAFF and BAFF-R in KOA patients Before treatment,the BAFFand BAFF-R two groups of KOA patients was no significant difference(P> 0.05). After treatment, the two groups of BAFF(%) and BAFF-R has decreased, the improvement of BAFFin the treatment group is better than the control group.3.2 Experimental Results3.2.1 The impact of Xinfeng Capsule on body weights, Mankin score of KOA rats Compared with the NC group, body weight of MC group significantly decreased,Mankin score was significantly increased(P <0.05 or P <0.01). Compared with the MC group, body mass of GS group was increased, Mankin score was significantly lower(P<0.05); XFC group significantly increased body weight, Mankin score was significantly lower(P <0.05 or P <0.01). Compared with the GS group, body weight of XFC group rats was significantly higher(P <0.05).3.2.2 Impact of Xinfeng Capsule on chondrocytes ultrastructural in KOA rat Compared with the NC group, MC group chondrocytes irregular shape, nuclear condensation, and even the disappearance of cytoplasmic rough endoplasmic reticulumincrease, expansion, mitochondrial degeneration seen,a large number of lipid droplets appear on the cell surface microvilli like projections significantly reduced, sparse matrix fibrils. Compared with the MC group, the treatment group had some improvement in cartilage cell ultrastructure. Compared to the treatment groups,GS group cartilage less structured nucleus,partial side, increased cytoplasmic rough endoplasmic reticulum,some expansion,the visible part of swelling of mitochondria,the cell filaments or fine particulate matter reduction. Visible micro cell surface villous projections matrix fibrils arranged denser,thicker,individual regions still sparse. XFC group most chondrocytes normal morphology,nuclei were oval,within a moderate amount of cytoplasm within the rough endoplasmic reticulum,a small number of mitochondria,lysosomes,a small amount of lipid droplets,some cells cytoplasm see more filamentous structure,the matrix fibril arrangement denser,thicker,and the NC group similar.3.2.3 Impact of Xinfeng Capsule on immunoglobulin in KOA rats Compared with the normal group,model group,serum Ig G1,Ig G2 a lower,(P <0.05 or P <0.01); compared with the model group, the Xin Feng capsule group Ig G1, Ig G2 a lower(P <0.05 or P <0.01).3.2.4 Changes of autophagy signaling of cartilage, thymus,spleen in KOA rat Xinfeng Capsule impact on them Compared with normal rats, Atg5 lower(P <0.05 or P <0.01) in rat spleen and thymus in OA cartilage, Atg7 lower(P <0.05 or P <0.01) in the spleen, thymus cartilage; m TOR increases in cartilage(P <0.05 or P <0.01), PI3 K, Akt were increased(P <0.01) in cartilage; LC3-II, Beclin-1 reduced cartilage.4. Conclusion4.1 KOA patients with elevated Ig G, wherein the elevated Ig G1, Ig G2, Ig G3 levels,(P <0.01), described in KOA patients,humoral immunity does participate in lesionst.4.2 Immunoglobulin changes in KOA patients are associated with joints and systemicsymptoms and signs physiological functions, anxiety and depression, disease activity,laboratory parameters(Ig A,Ig M,CRP,ESR,etc.) are also closely related. Description KOA only cause joint swelling,pain,deformation,flexion and extension negative,but can also lead to reduced immune globulin, and the emergence on the basis of physiological, psychological and overall systemic dysfunction.4.3 While reduce pain, joint swelling and other localized symptoms of KOA patients,Xinfeng Capsule can reduce the immune globulin, regulating systemic humoral immune response and to an overall improvement in less gas lazy words, loss of appetite and other symptoms, reduce CRP, ESR and other inflammatory activity index.4.4 Mechanism of Xinfeng Capsule improve the KOA patients.4.4.1 Immune-inflammatory, inhibiting damage pathological of cartilage Xinfeng Capsule can significantly reduce the KOA rats Mankin score, improving cartilage cell ultrastructure, indicating XFC cells by regulating the immune response,reduce joint inflammation locally, microcirculation, increase local blood supply and oxygen supply, so that improved joint function.4.4.2 Raised inflammatory cytokine suppression, reduced proinflammatory cytokines,correct imbalance of synthesis of cartilage cells4.4.3 Adjust BAFF, lower humoral4.4.4 Regulate the expression of PI3 K / Akt-m TOR signal, improving metabolism, to restore homeostasis4.4.5 Adjust Beclin-1 signal transduction pathway, improve autophagy state...