Mechanism Research Of Xushi Yangyin Ruzao Recipe On Sjogren Syndrome By NF-?B Signaling Pathway Mediated By BAFF/BAFF-R

1 purposeTo observe the changes of clinical symptoms,clinical indexes,B lymphocyte function,cytokines and BAFF/BAFF-R/NF-?B signaling pathway in patients with primary Sjogren's syndrome?pSS?before and after treatment.To analyze the changes of B lymphocyte function in patients with pSS and its correlation,to evaluate the clinical efficacy of Xushi Yangyin Ruzao Recipe in treating pSS,and to explore the mechanism of Xushi Yangyin Ruzao Recipe in treating pSS from the point of view of NF-?B signal pathway mediated by BAFF/BAFF-R.To provide a theoretical basis for the treatment of pSS,broaden the idea of pSS treatment,guide clinical use of drugs.2 methodSixty patients with primary Sjogren's syndrome from rheumatology department of traditional Chinese medicine hospital in Anhui province were divided into two groups according to random numerical table:Xushi Yangyin Ruzao Recipe group and TGP control group,from December 2017 to December 2018.There were 30 cases in each group.In the same period,20 normal persons in the Health Management Center of Anhui traditional Chinese Medicine Hospital were selected as the healthy control group.60 cases of TCM syndrome,disease activity,quality of life,anxiety and depression were evaluated by questionnaire.Experimental indexes and methods:blood routine indexes were detected by blood cell analyzer;the absolute counts of B cells,the percentage of B10(CD19⁺CD1d^hiCD5⁺)and the expression of BAFFR on the surface of B cells were detected by flow cytometry.The levels of IL-21,IL-10,BAFF,NF-kB and TNF-?were detected by ELIAS,the levels of RF,CRP,IgM,IgA,IgG,C3 and C4 were detected by biochemical analyzer;ESR was detected by ESR analyzer.The ESSDAI score was calculated according to the objective systemic symptoms.3 results3.1 Changes of B cell function,cytokines and BAFF/BAFF-R/NF-?B signaling pathway in patients with pSS.Compared with NC group,the expression levels of B%,ESR,RF,IgG,IL-21,BAFF,NF-KB and TNF-?in peripheral blood of patients with pSS were significantly higher than those in the control group?P<0.05?.The expression levels of CD19⁺CD1d^hiCD5⁺,C3,IL-10 were significantly decreased?P<0.01?.3.2 Study on the correlation of B cell function in patients with pSSThe correlation analysis showed that the absolute counts of peripheral blood B cells in pSS patients were positively correlated with the level of RF,IgG,IL-21,BAFF,and negatively correlated with the levels of C4,IL-10 and lacrimal film rupture time?P<0.05?.B%was positively correlated with the level of ESR,IgG,RF,IL-21,BAFF,NF-kB,TNF-?,and negatively correlated with C4,IL 10,static saliva flow rate and lacrimal film rupture time?P<0.05?,while CD19⁺CD1d^hiCD5⁺%was positively correlated with C4,IL-10 level?P<0.05?.There was a positive correlation between the rate of static saliva flow and the time of lacrimal filmrupture,andanegativecorrelationwith ESR,IgG,IL-21,BAFF,NF-kB,TNF-?and corneal fluorescence staining?P<0 05?.The score of symptom,ESSDAI score,total score of quality of life and its subcategories?social function,psychological function score?were positively correlated with the absolute count of B cells.Total integral of TCM syndrome and its subcategories?dry eyes,tongue,oral mucosa,irritating heat sweating,salivary gland swelling,dry mouth and pharynx,tiredness?,ESSDAI score,total quality of life score and its subcategories?psychological function,social function score?and anxiety and depression scale score were positively correlated with B%expression level?P<0.05?,and the scores of fatigue symptom score,total quality of life score and its subcategories were positively correlated with B%expression level.The total score of TCM syndrome and its subcategories?dry eyes,tongue,oral mucosa,salivary glands,fatigue,dry mouth and pharynx,symptoms of irritating heat and night sweating?,ESSDAI score,psychological function,etc.There was a negative correlation between anxiety scale score and CD19⁺CD1d^hiCD5⁺%expression level?P<0.05?.3.3 Changes of therapeutic efficacy and observation indexes in two groups of patients with pSS3.3.1 comparison of therapeutic effects between two groups of pSS patientsAfter treatment,the total effective rate,obvious effective rate and effective rate of the treatment group were 86.7%,56.7%and 30.0%respectively,and the total effective rate,effective rate and effective rate of the control group were 76.7%,40.0%and 36.7%respectively.Compared with the control group,the effective rate and total effective rate in the treatment group were significantly higher than those in the control group?P<0.05?.Compared with pre-treatment,the total score of TCM syndrome and its subcategories in the treatment group(dry eyes,dry mouth,dry skin,tongue,oral mucosa,irritating heat sweating,constipation,dry nasal cavity,dry vagina,joint pain,etc.The scores of symptoms of fatigue and fatigue,total score of quality of life and its subcategories?social function,psychological function and self-health awareness ability?and scores of anxiety and depression scale were significantly decreased?P<0.05?.In the control group,the subtypes of TCM syndrome?dry eyes,dry mouth,dry throat,constipation,dry symptoms of nasal cavity?,the total score of quality of life and its subcategories?social function,self-health awareness ability?were significantly decreased?P<0.05?.Compared with the control group,the total integral of the TCM syndrome of the treatment group and its subclasses?dry and astringent,dry and dry throat,tongue and mouth,oral mucosa,heat and night sweat,dry vaginal dryness,and asthenia of the body?,total integral of the quality of life and its subclasses(social function,psychological function,The scores of self-health recognition ability and anxiety and depression scale were significantly lower?P<0.05?.3.3.2 comparison of observation Indexes between two groups of patients with pSSThe levels of absolute B cell count,B%,ESR,RF,IgG,ESSDAI,IL-21,BAFF,NF-kB?P52?,TNF-?and corneal fluorescence staining in the treatment group were significantly lower than those before treatment?P<0.05?,CD19⁺CD1d^hiCD5⁺%,C3,IL-10,The static saliva flow rate and lacrimal film rupture time were significantly higher than those before?P<0.05?.In the control group,the levels of B%,ESRs,IgG,IL 21,BAFF,NFKB,TNF?and corneal fluorescence staining were significantly lower than those before.The levels of CD19⁺CD1d^hiCD5⁺%,C3,IL-10,static saliva flow rate and lacrimal film rupture time were significantly higher than those before?P<0.05?.Compared with the control group,the levels of B%,RF,IgG,IL-21 and BAFF in the treatment group were significantly lower than those in the control group?P<0.05?.The levels of CD19⁺CD1d^hiCD5⁺%,C3,IL-10,static saliva flow rate and lacrimal film rupture time in the treatment group were significantly higher than those in the control group?P<0.05?.4 conclusion4.1 The absolute count of B cells,the increase of B%level and the decrease of B10 level in patients with pSS indicated that the B cells were in abnormal activation state in pSS patients.4.2 The expression of BAFF,NF-?B in pSS patients was significantly increased,suggesting that the BAFF/BAFFR-NF-?B signaling pathway was in an overactive state,and the correlation between B cell function and signal pathway parameters was analyzed.The results suggest that the abnormal B cell function in pSS patients is related to the excessive activation of BAFF/BAFFR-NF-?B signaling pathway.The scores of TNF-?,IL-21,ESR,RF,IgG,ESSDAI,TCM syndrome integral,quality of life,anxiety and depression in PSS patients are increased.And the correlation between B-cell function and B-cell function showed that it was related to abnormal activation of B cells.4.3 Xushi Yangyin Ruzao Recipe is effective in the treatment of SS,and the total effective rate is slightly higher than that in the control group.It can significantly improve the scores of TCM syndrome,local symptoms of mouth and eye,quality of life,anxiety and depression.The activation level of B cell and BAFF/BAFFR-NF-?B signaling pathway was significantly decreased.4.4 Xushi Yangyin Ruzao Recipe can down-regulate the level of BAFF/BAFF-R,inhibit the activity of NF-?B signaling pathway,inhibit the abnormal activation and proliferation of B cells,regulate the differentiation of B cell subsets,and upregulate the proportion of regulatory B cells?B10?.It can promote the production of anti-inflammatory factor IL-10,inhibit the production of pro-inflammatory factor IL-21,inhibit the excessive abnormal growth,proliferation and activation of B cells,so as to inhibit the status of glandular immune inflammation and achieve the purpose of improving the condition of the disease.