Research Of The Syndrome Characteristic Of Osteoarthritis Of Knee And The Relativity With Serum Biomarker CTX-Ⅱ And COMP

ObjectiveOsteoarthritis of the Knee (KOA) is a highly incident rate disease. At the present time, To diagnosis this disease is mainly according to symptoms, X-ray and CT, etc. they could have only morphological abnormalities changes when articular cartilage had serious injury, But at this period most patients had missed the best time to cure it and damaged the quality of lives serious. In recent years, within the continuous development of molecular biology, There is a hot research to reflect early degeneration in articular cartilage and other relevant indicators from pathology laboratory.Therefore, It is the ultimate purpose to investigate biomarkers CTX-Ⅱand COMP of clinical significance from cartilage metabolism in knee osteoarthritis patients, to research of syndrome characteristic in TCM and the relativity with biomarkers CTX-Ⅱand COMP. At the same time, we will combine with Cluster analysis of Traditional Chinese Medicine syndrome type and experimental way of zoology to make our results more objective and reliable.MethodsFirst, we designed questionnaires according to the clinical manifestations of patients with knee osteoarthritis, and collected to meet the requirements of 70 patients which including about clinical symptoms and signs, tongue, pulse and other diagnostic messages, and combined with cluster analysis statistical method and clinical expertises,we hope to explore and sum up the disease characteristics of syndromes and to provide objective foundation for clinical research;Secondly, one experimental group included 70 patients who had detailed informations and met the inclusion criteria, another comparison group included 18 persons who had no medical history of knee joints and had normal physical examination and no medical history of metabolic disease. Two groups of serum biomarkers CTX-Ⅱand COMP levels were detected by Enzyme-linked immunosorbent assay(ELISA)method. Then we used SPSS statistical software to analyze the data from the experimental group and comparison group, and from different gender, age, X-ray levels and different syndromes between CTX-Ⅱand COMP levels change. Finally, To link with experimental animal studies, we used Pearson statistics to find the relationship between content of CTX-Ⅱand COMP and Mankin scores in different weeks from articular cartilage degeneration of mice, so that we hope to find the correlation among serum content of CTX-Ⅱ,COMP and cartilage degeneration in intuitionistic and profound.Results(一)Cluster analysis using syndrome of TCM in KOAFirst of all, we analyzed frequence and frequency from the four diagnostic symptoms and removed the frequency which less than 5%,Finally we got the twenty-nine symptoms as indicators of Chinese medical observation. The four diagnostic symptoms frequency table was summarized, we could see the majority indicator was asthenia syndrome in Degenerative Knee Osteoarthritis, and the frequencies from high to low order were joint dull pain (54.29%),fatigue increased (51.43%),waist and knee debility(50%),encounter cold pain(44.29%),weak and thin pulse(41.43%)by followed. Then we used Ward method of Hierachical Cluster Analysis to Observe the classification from different syndromes when they were clustered four class,three class and two class, At the same time we used the cluster tree diagram to research. When the cluster tree diagram from the rescaled distance cluster was fifteen, the twenty-nine symptoms which we just mentioned about could be divided into three categorie syndromes:one kind of thermophilic contained overfatigue,weak and thin pulse,joint deformity,tongue reddish,joint dull pain,thin white moss,limbs not warm, clear urine long,waist and knee debility,constipation,hands and feet deficiency-hot, they belonged to Deficiency of liver and kidney in syndrome of TCM; Another kind of thermophilic contained chills,cold,pale tongue and face, they belonged to arthralgia caused by wind,cold and dampness in syndrome of TCM;The third class included aggravated pain at night,purple tongue,gray face,joint irritation,thirst but not drink, they belonged to blood stasis and vital energy retardation in syndrome of TCM. Finally, we found from cluster pie that cluster three maked four diagnostic data with good dispersion, clear distribution in symptom and the most fit for clinical practice. So the disease could be divided into three categories syndrome:The first symptom was Deficiency of liver and kidney; The second symptom was arthralgia caused by wind, cold and dampness;The last symptom was blood stasis and vital energy retardation.the frequency from those three symptoms were different, Deficiency of liver and kidney Occupied 41.43%,arthralgia caused by wind, cold and dampness Occupied 35.71%,blood stasis and vital energy retardation Occupied 22.86%. The highest rate was Deficiency of liver and kidney.(二)Clinical study about KOA with biomarkers CTX-Ⅱand COMP(1)In patients of KOA, the biomarker CTX-Ⅱwas about (1057.03±195.05, pg/ml) and COMP was about (557.02±85.44, pg/ml), In comparison group the serum biomarker CTX-Ⅱwas about (672.78±148.67, pg/ml) and COMP was about (353.01±54.13, pg/ml), two groups of biomarkers had significantly difference(P< 0.05), the experimental group data were higher than the comparison group.(2) From different gender variation study we found that the serum biomarker CTX-Ⅱin male patients was about (975.09±172.88, pg/ml) and in female was about (1094.58±194.72, pg/ml), two groups of CTX-Ⅱhad significantly different too(P<0.05), the conlusion was that female group was higher than male group. But we found no significant difference in COMP between male and female patients (P>0.05). In less than 50-year-old age change groups, we found that there were no different distinction between men and women in CTX-Ⅱand COMP. Serum CTX-Ⅱand COMP were significantly hoist in female patients (50-59 age), and had significantly different(P< 0.05)when compared with the former group, however after 60 age, CTX-Ⅱbegan to get down,but COMP was opposite;After 50 age of male patients the CTX-Ⅱand COMP continued to increase, there was no significant difference in CTX-Ⅱ(P> 0.05), but COMP still had significantly different(P<0.05);When female patients from 50 to 59 age, we thought that was the time of menopause, great changes in hormone levels, bone degeneration obvious, and thus a greater impact on the metabolism of articular cartilage, All of this indicate that there was a peak incidence, so it had an important guiding significance to prevent and treat this disease.(3)In the levels of X-ray study, we found that there were significantly difference between levelⅠ,Ⅳand levelⅡ,Ⅲin patients' serum biomarker CTX-Ⅱ(P< 0.05),and there were also significantly difference between levelⅠand other levels in COMP, but the other three groups showed no significant difference(P> 0.05). The level of serum CTX-Ⅱin the early stage increased not obviously, and when joint space narrowing it reached the higest level, then when the joint space reducing it decreased again,There were no completely tendency between COMP and CTX-Ⅱin our study, we thought that synthesis and degradation of COMP might be also related to pathological changes in synovial inflammatory.(4) In different syndromes of TCM, we found that wind,cold and dampness syndrome was obvious lower than deficiency of liver and kidney and blood stasis and vital energy retardation syndrome in CTX-Ⅱ, and had statistically significant difference (P< 0.05), Although COMP had trendence to upward, there was no significantly difference among them(P> 0.05), which might indicate that there might be exist some different between degeneration of cartilage and synovial reaction in difference types of TCM, So that we hope CTX-Ⅱand COMP may become an objective basis for micro-differentiation of TCM.(三)The correlation study about CTX-Ⅱand COMP with cartilage degeneration in animal modelsThere was statistically significant difference between experimental group and normal contrast group in levels of CTX-Ⅱand COMP(P< 0.05), However, they had different increased time. At the beginning of two weeks, there was no significant difference level in CTX-Ⅱ, following the time past, there was significant difference in four week(P< 0.05), especially in six week(P< 0.01). There was significant difference from two week and four week in level of COMP(P< 0.05)and increased obviously at the six week. We found correlation between CTX-Ⅱand COMP with cartilage degeneration that CTX-Ⅱwith OA had most high positive correlation, r=0.923, the second was COMP with OA, r=0.899, the relevance of them had significant linear relationship by the correlation coefficient tests of two-tailed (P< 0.05).Conclusion(1) During this reseach, We find the most symptom type of TCM in Knee Osteoarthritis is Deficiency of liver and kidney, the second is wind, cold and dampness, the last symptom is blood stasis and vital energy retardation. So it prove that Cluster analysis using in symptom type of TCM in KOA has some scientific, practical and credible.(2)There have closely related relationship between CTX-II and COMP in primary knee osteoarthritis, If together them can greatly improve the accuracy of diagnosis, so they play an important role in the pathogenesis of knee osteoarthritis and have close relationship with cartilage degeneration and can reflect the condition and syndrome of TCM.