The Correlations Between TCM Syndrome Type And Inflammatory Cells Factor, Bone Metabolism Of Index Correlation In Patients With Ankylosing Spondylitis

Objective: Ankylosing spondylitis (AS) is a chronic, progressiveinflammatory autoimmune disease, which is characterized by maininvolvement of sacroiliitis and axial joints system,but sometimes peripheraljoints,tendons attachment points are involved to some degree. It’s the mainpathological characteristics are the hyperplasia of synovial cells, Lymphocyteinfiltration and the formation of pannus, thus cause bone and cartilage erosiondamage, spine, tendons and ligaments around the place such as the bonystiffness of the disc and is accompanied by local osteoporosis. The etiologyand pathogenesis is still not clear, the prevalence rate of around0.3%in China,of which about60%of the patients hip involvement, the involvement of thespine happen for osteoarthritis stiffness, lead to disability. Bone destructionand new bone formation are the main reason for AS disability. In recent years,research shows that osteoclast(OC) plays an important role in bone destructionand osteoporosis pathological mechanism.And the imbalance betweenosteoblast and osteoclast plays a decisive role in the calcium homeositasis. Inthe whole process of bone damage of inflammatory joint disease, somepromote inflammation factors can’t have direct action on the differentiationand activation of OC, but rather have an indirect effect through regulating theexpression of members of the tumor necrosis factor (TNF) family such asreceptor activator of nuclear factors κ B ligand(RANKL), receptor activatorof nuclear factors κ B(RANK), osteoprotegerin(OPG). RANKL, RANK, OPGare indispensable molecules of adjusting osteoclast activity, which were themost important factors in the whole process of bone damage of inflammatoryjoint disease. The canonical Wnt signaling pathway is important to jointremodeling in which Dickkopf-1(DKK-1) and sclerostin(SOST) are master regulator.The aim of this study is to test the levels of peripheral blood serummacrophage migration inhibitory factor(MIF), Interleukin-23(IL-23), RANKL,OPG and Dickkopf-1(DKK-1), Sclerostin(SOST) in patients with active AS,and with active rheumatoid arthritis (RA) and healthy controls forcomparison.To understand the correlation of the inflammatory cytokine MIF,IL-23and bone destruction factors/bone remodeling, disease activity, andChinese Medicine Syndrome Type (TCM) is correlated with inflammatorycells factor, bone destruction/bone remodeling related factors, which tries toexplore further the the pathogenesis of inflammation and bone destruction inactive AS so as to seek new therapeutic targets of improving the bonemetabolism and delaying for bone destruction.In order to not only obtain theexperiment-based evidences of AS Chinese medicine diagnosis but alsoexplore a new idea and method for diagnosis and treatment of traditionalChinese and western medicine in AS.Methods: Choosing30hospitalized patients with active AS as defined bythe New York criteria were investigated as study group.Meanwhile the controlgroup consisted of30hospitalized patients with active RA(as defined by theACR/EULAR RA criteria and DAS28rating criteria) and15healthyvolunteers. The group of patients with active AS were divided according toTCM syndrome type. Recording the age, gender, duration, erythrocytesedimentation (ESR) and C-reactive protein (CRP) of patients with RA or ASrespectively.The ESR was determined according to the Westergren method.CRP levels were measured with a Behring nephelometer and Behring reagents.Choosing Bath ankylosing spondylitis disease activity index(BASDAI) andDisease activity score(DAS28) as the evaluate indexes of diseaseactivity,respectively.Blood samples were collected using a standardvenepuncture technique in the early morning.Serum was separated.By enzymelinked immunosorbent assay, the levels of serum MIF,IL-23,RANKL,OPG,DKK1and SOST were detected,to compare the expression ofMIF,IL-23, RANKL,OPG,DKK1and SOST in different patient groups and normal subjects.In addition，to analyze the correlation of inflammatory cellfactors and the cytokines of bone metabolism,and further to analyze thecorrelation of the aboved cytokines and the clinical indexes,TCM.DataStatistics: The results were expressed as±s. All statistical analyses of datawere computed by Statistical Package for Social Sciences (version13.0). Ifmeasurement data,the varience is equal,the comparisons of mean amonggroups were evaluated by one-way analysis of variance (ANOVA);if not bynonparametric test for several independent samle. Pearson correlated-test wasadopted in correlation analysis. Count data was used a chi-square test.P <0.05or P <0.01was all judged as a significant difference.Results:1The general informationAS group:25male,5female; age (18-58) years, the mean age(29.43±8.33) years; disease duration (0.3-20) years, the mean disease duration(6.73±4.18)years; RA group:5male,25female; age (15-75) years, the meanage(46.07±14.79)years; disease duration (0.2-16) years, the mean diseaseduration(4.99±5.15)years; Normal control group3male,12female, the meanage(30.20±5.20).ASgroup:BASDAI(3.94±1.65);ESR(40.25±29.79)mm/h;CRP(4.78±3.40)mg/dl;RAgroup:DAS28(6.06±1.25);ESR(69.79±27.46)mm/h;CRP(4.31±4.25)mg/dl.2Comparison serum IL-23(pg/ml)The level of serum IL-23in AS group (74.90±42.24) was higher thannormal control group (43.32±16.06), but less than RA group(146.60±117.46).There were statistical differences among AS group, RAgroup and normal control group (P<0.05). Compared with RA group, therewas no statistically difference (P>0.05).3Comparison of serum MIF(ng/ml):The level of serum MIF in AS group (25.38±11.05) was higher thannormal control group (20.58±6.44), but less than RA group(37.70±14.19).There were statistical differences among AS group, RA group and normal control group (P<0.05). Compared with RA group, there werestatistically differences (P <0.05).4Comparison of serum RANKL(pg/ml):The level of serum RANKL in AS group (21.53±17.44) was higher thannormal control group (8.39±5.88) and RA group (12.12±9.72).There was nostatistical differences among AS group, RA group and normal control group(P>0.05). Compared with RA group, there was no statistically differences (P>0.05).5Comparison of serum OPG (pg/ml):The level of serum OPG in AS group (5.04±1.65) was higher than normalcontrol group (3.39±1.02) but less than RA group (8.48±3.61).There werestatistical differences among AS group, RA group and normal control group(P<0.05). Compared with RA group, there were statistically differences(P<0.05).6Comparison of serum DKK1(ng/ml):The level of serum DKK1in AS group (0.71±0.16) was higher thannormal control group (0.66±0.20) and RA group (0.70±0.28).There was nostatistical differences among AS group, RA group and normal control group(P>0.05). Compared with RA group, there was no statistically differences (P>0.05).7Comparison of serum SOST(ng/ml):The level of serum SOST in AS group (1.39±0.53) was higher thannormal control group (0.98±1.02) and RA group (1.43±0.55).There was nostatistical differences among AS group, RA group and normal control group(P>0.05). Compared with RA group, there was no statistically differences (P>0.05).8Correlations among the clinical disease activity indexes and the levelsof serum IL-23,MIFThere was no correlation among the level of serum IL-23and diseaseduration,ESR,CRP,BASDAI(r=-0.142,0.088,-0.018,-0.421,respectivelyP>0.05) in AS group. There were significant positive correlations among the level of serum MIF and disease duration(r=0.496,P<0.01), but there was nocorrelation among the level of serum MIF and ESR, CRP, BASDAI (r=0.361,0.299,0.270,respectively,P>0.05) in AS group.9Correlations among the bone metabolism indexes and the levelsofserum IL-23,MIF and the clinical disease activity indexesThere was no correlation among the level of serum RANKL and diseaseduration,ESR,CRP,BASDAI,IL-23,MIF,OPG,DKK1,SOST(r=-0.116,0.003,0.147,0.069,-0.008,0.314,0.119,-0.199,0.163,respectively,P>0.05).There weresignificant positive correlations among the level of serum OPG and diseaseduration(r=0.520, P<0.01), and there were positive correlations among thelevel of serum OPG and ESR(r=0.379, P<0.05)，but there was no correlationamong the level of serum OPG andCRP,BASDAI,IL-23,MIF,DKK1,SOST(r=0.338,0.173,0.379,0.759,0.099,0.252,respectively,P>0.05). There were positive correlations among the level ofserum DKK1and BASDAI (r=0.414, P<0.05), but there was no correlationamong the level of serum DKK1and disease duration, ESR, CRP, MIF,SOST(r=0.128,0.099,-0.013,-0.105,0.087,respectively,P>0.05). There was nocorrelation among the level of serum SOST and diseaseduration,ESR,CRP,BASDAI,IL-23,MIF(r=-0.242,0.266,0.354,-0.206,0.020,0.200,respectively,P>0.05).10The distribution of AS TCM syndrome, and the relationship betweenclinical-related indexes,the level of serum IL-23,MIF andRANKL,OPG,DKK1,SOST10.1The distribution of AS TCM syndrome:The percents of five TCM syndrome as follow:the retention of damp-heatsyndrome10%(3/30), the retention of damp-cold syndrome27%(8/30),theretention of stagnated blood syndrome47%(14/30),the deficiency ofkidney-yang syndrome10%(2/30) and the deficiency of liver-kidneysyndrome10%(3/30).Active AS is mainly about the retention of stagnatedblood syndrome.10.2The relationship between TCM syndrome type and age, disease duration10.2.1The age of deficiency liver-kidney syndrome ismaximum(40.00±9.17),but the retention of damp-heat syndrome isminimum(27.33±10.69).There was no significant differences in eachsyndrome (P>0.05).10.2.2The disease duration of deficiency liver-kidney syndrome ismaximum(14.33±1.53), but the retention of damp-heat syndrome isminimum(4.33±2.47).There were significant differences in each syndrome(P<0.01).10.2.3There was no significant differences in each syndrome(P>0.05).10.3The relationship between TCM syndrome and ESR,CRP,BASDAIThere was no statistical differences about ESR,BASDAI in eachsyndrome (P>0.05). There were statistical differences about CRP in eachsyndrome (P<0.01).10.4The relationship between TCM syndrome and IL-23,MIFThere were statistical differences about the level of serum IL-23in eachsyndrome (P<0.01).The level of serum IL-23was highest(112.20±57.68) inthe retention of damp-cold syndrome. There was no statistical differencesabout the level of serum MIF in each syndrome (P>0.05).10.5The relationship between TCM syndrome andRANKL,OPG,DKK1,SOSTThere were statistical differences about the level of serum SOST in eachsyndrome (P<0.05). But there was no statistical differences about the level ofserum RANKL,OPG,DKK1in each syndrome (P>0.05).Conclusions:1There are bone metabolic abnormalities in active AS. The levels ofserum RANKL,OPG are rising, and with the extension of disease course,new bone formation plays a main role in the process of the change of jointstructure. IL-23, MIF may be mediated indirect inflammation reaction in bonemetabolism in AS disease process.2There is no correlations of the levels of serum DKK1, SOST and ESR, CRP, BASDAI, IL-23, MIF in active AS, which suggest DKK1, SOST mayparticipate in the progress independently.It provides new targets for furtherresearch on interventing AS bone metabolism mechanisms.3In the distribution of AS TCM syndrome, active stage is mainly aboutthe retention of stagnated blood syndrome.There were significant differenceson duration、CRP、IL-23、SOST between syndromes. There was no correlationof the sacroiliac joint imaging classification and TCM syndrome type.Modernclinical and laboratory relevant quantitative/qualitative indexes mey be act asobjective evidence of TCM syndrome differentiation in active AS.