目的:通过检测关节软骨Sox9、Col2al基因在阳虚质与平和质两者之间的表达水平,探讨膝骨性关节炎中Sox9、Col2al与中医体质的内在联系和规律.方法:2012年1 0月20日至2014年12月27日,对甘肃省中医院关节骨科门诊及住院部确诊为膝骨性关节炎的患者进行中医体质问卷调查,同时调查、收集非膝骨性关节炎正常人群的一般资料.在显微镜下观察不同体质KOA患者软骨的病理改变,采取免疫组化及Real-Time RT-PCR的方法检测Sox9基因、Col2al在阳虚质膝骨性关节炎患者膝关节软骨细胞mRNA及蛋白水平中的表达.结果:在对实验结果进行分析后发现在不同体质的KOA患者中,阳虚质KOA患者中软骨内细胞排列紊乱,细胞聚集成团或者局部退化消失,Sox9在软骨细胞核中表达,Col2al在细胞外基质中表达,在阳虚质KOA、阳虚质非KOA患者软骨细胞内呈现高表达状态,在平和质KOA相对阳虚体质表达比较低,两者的表达比较差异均有统计学意义(P＜0.05).结论:膝骨性关节炎发病的体质危险因素主要为阳虚质.在分子表达水平Sox9及Col2al均呈现高表达,其与中医体质存在一定的内在相关性,说明体质因素是诱发膝骨性关节炎的高危因素,因人的体质是相对恒定的,某一疾病的诱发是具有时间性的,故阳虚质是膝骨性关节炎中诱导Sox9及Col2al高表达的主要因素之一.%Objective:To discuss the internal relations and rules of Sox9,Col2al and traditional Chinese medicine constitutions in knee osteoarthritis through detecting the expression level of Sox9 and Col2al genes in the patients of Yang deficiency and constitution.Methods:From October 20th,2012 to December 27th,2014,the TCM constitution questionnaire for patients in outpatient and inpatient in joint department of orthopedics of Gansu Provincial Hospital of traditional Chinese medicine with diagnosis of knee osteoarthritis was conducted,at the same time,the general information of non knee osteoarthritis and normal people were investigated and collected.The cartilaginous pathological changes of KOA patients in different constitutions under the microscope was observed,and the expressions of Sox9 gene and Col2al in mRNA of knee cartilage cells and the protein of Yang deficiency patients with osteoarthritis were detected with the methods of immunohistochemistry and Real-Time RT-PCR.Results:After the analysis of the experimental results,it was found that in the KOA patients with different physical constitutions,the arrangement of the chondrocytes were disordered in KOA patients with Yang deficiency,and the cells were aggregated into clusters or local degeneration and disappearance.Sox9 was expressed in the nucleus of cartilage while Col2al was expressed in the extracellular matrix.They were highly expressed within the cartilage cells in KOA patients with Yang deficiency and non KOA patients with Yang deficiency,and the expression of KOA patients with normal constitution was relatively lower than that of KOA patients with Yang deficiency.There was a significant difference between the two expressions (P＜0.05).Conclusion:The main risk factors of the morbidity of knee osteoarthritis is Yang deficiency constitution.At the molecular expression level,Sox9 and Col2al are both highly expressed,and there is a certain intrinsic correlation with TCM constitution.This shows that the constitution factors are the high risk factors which can induce knee osteoarthritis.People's constitution is relatively constant,the induction of a disease is a matter of time.Therefore,Yang deficiency constitution is one of the main factors which induce Sox9 and Col2al high expression in knee osteoarthritis.
展开▼
