A Study On The Correlation Of PKC、β-catenin And Mmp-13 In The Articular Cartilage Of The Knee Osteoarthritis Patients With Yang Asthenia Constitution

Objective: This paper aims to apply the clinical epidemiology method to investigate the constitution type distribution characteristics of traditional Chinese medicine and related pathogenic factors of knee osteoarthritis. Based on the medical discovery that Yang asthenia constitution has a high incidence of KOA, this paper probes into the correlation of PKC, β-catenin and MMP-13 in the articular cartilage of the knee osteoarthritis patients with Yang asthenia constitution at the molecular level with the aid of immunohistochemistry and RT-PCR detection method, makes a thorough analysis of the related pathogenesis of the knee osteoarthritis patients with Yang asthenia constitution in northwest area, to provide an objective basis for the diagnosis of this disease and a new idea and therapeutic target for the prevention and treatment of this disease.Methods: Applying the clinical epidemiology method, this paper refers to the standard of Classification and Determination of Constitution in TCM to make “Yang asthenia constitution KOA and Wnt/beta-Catenin signaling pathway research” and the research questionnaire is designed uniformly. KOA patients with TCM constitution are determined, specimens of articular cartilage are collected and the database of KOA patients with TCM constitution is established. There are 20 cases of KOA patients are selected randomly from the database, including 10 cases of non-Yang asthenia constitution and 10 cases of Yang asthenia constitution. Another 10 cases of normal non-KOA patients are regarded as the control group. Literature evidence-based method is used to choose the genes which are closely related to articular cartilage metabolism(PKC, β-catenin and MMP-13) in KOA. Based on the molecular biology, this paper detects the expression of the three genes in different groups with the methods of immunohistochemistry and RT-PCR, and analyzes the expression of the three genes between KOA and normal control group, Yang asthenia constitution KOA and normal control group, non-Yang asthenia constitution and Yang asthenia constitution, non-Yang asthenia constitution and normal control group.Results:(1) according to the survey of 406 cases of KOA, Yang asthenia constitution,Qi asthenia constitution and Qi stagnation constitution are common in KOA patients.There are 164 cases of Yang asthenia constitution, accounting for 40.39%, 117 cases of Qi asthenia constitution, accounting for 28.82%, 65 cases of Qi stagnation constitution, accounting for 16.01%.(2) As immunohistochemical study shows, PKC and MMP-13 are expressed in normal articular cartilage, Yang asthenia constitution KOA and non-Yang asthenia constitution KOA, and the expression is mainly located in the cytoplasm and nucleus. A significant difference can be found when making a pairwise comparison between groups(P<0.05), which takes on a statistical significance. β-catenin expression in normal control group of articular cartilage is negative, but not in Yang asthenia constitution KOA and non-Yang asthenia constitution KOA. The expression is mainly in the nucleus. The comparison between two cases has no significant difference(P>0.05).(3) RT-PRC study shows:there is obvious statistical significance when PKC and MMP-13 genes are comparedin the normal control group and the articular cartilage tissues of KOA cases(P<0.01).The difference is statistically significant when PKC and MMP-13 genes are compared in pairs in Yang asthenia constitution cases, non-Yang asthenia constitution cases and control groups(P<0.05). β-catenin gene has not significant expression in the normal control group while it takes on a significant difference when it is compared with the expression of the articular cartilage tissues of KOA cases. The difference has not statistical significance when β-catenin gene is compared between Yang asthenia constitution cases and non-Yang asthenia constitution cases(P> 0.05).Conclusions:(1)this paper reconfirms that Yang asthenia constitution has a high incidence of KOA.(2)PKC gene has a correlation with the incidence of KOA, and may also has a intimate correlation with the high incidence of Yang asthenia constitution KOA.(3)β-catenin gene has a correlation with the incidence of KOA, but has no intimate correlation with the high incidence of Yang asthenia constitution KOA.(4)MMP-13 gene has a correlation with the incidence of KOA, and may has a intimate correlation with the high incidence of Yang asthenia constitution KOA.