原发性膝骨关节炎患者滑膜、滑液HMGB1表达与疾病严重程度的相关性研究

发布时间：2018-04-24 10:13

本文选题：膝骨关节炎 + 高迁移率族蛋白B-1　；参考：《新疆医科大学》2015年博士论文

【摘要】：目的:评估原发性膝骨关节炎(OA)患者临床症状、膝关节X线改变及两者之间的关系;观察膝OA患者膝关节滑膜细胞上高迁移率族蛋白B1(HMGB1)表达及分布情况,初步探讨滑膜细胞表达的HMGB1在膝OA疾病发展中的作用;分析膝OA患者滑液HMGB1水平与临床症状、滑膜病理改变之间的关系,探讨膝OA患者滑液HMGB1水平能否作为反映疾病严重程度的生物学标记物,为早期诊断膝OA提供新方法。方法:纳入满足标准的原发性膝OA患者74例,采用WOMAC评分量表评估手术前患者膝关节疼痛、僵硬及日常活动情况;采用KL分级评估膝关节X线改变的严重程度。膝OA组根据KL分级分为KL2/3组(中度病变组)和KL4组(重度病变组)两个亚组。对照组为34例同一时期因外伤引起交叉韧带断裂或半月板撕裂需要关节镜下治疗的患者,对照组无OA病史,膝关节影像学X检查正常。光镜下从衬里层增生、血管内膜下细胞密度和炎症细胞浸润三个方面对滑膜病变进行评分来评价滑膜炎严重程度;采用免疫组化技术观察HMGB1在滑膜细胞核内、外的分布情况,计算核外HMGB1染色阳性的滑膜细胞比例;采用蛋白印迹法(Western Blot)测定滑膜HMGBl蛋白相对水平;采用酶联免疫吸附法(ELISA)测定滑液HMGB1含量。运用SPSS21.0统计软件对实验结果进行统计学分析。结果:Spearman相关性分析发现KL分级与WOMAC总分及日常活动评分存在显著相关性,与疼痛和僵硬评分无显著相关性;HE染色显示:对照组30例和OA组4例滑膜为正常滑膜;对照组4例和OA组40例滑膜为轻度滑膜炎;OA组30例滑膜为重度滑膜炎,两组患者滑膜炎评分差异有统计学意义(P0.05)。滑膜免疫组化显示:对照组滑膜细胞HMGB1主要表达在细胞核内,而OA组滑膜细胞HMGB1则呈现出核内、外及胞外表达;进一步计数发现HMGB1表达阳性的滑膜细胞比例OA组高于对照组(42.5%vs 39.7%),差异有统计学意义(P0.05);核外HMGB1表达阳性的滑膜细胞比例OA组高于对照组(24.0%vs 5.7%),差异有统计学意义(P0.05);Western Blot和ELISA结果表明OA组滑膜和滑液HMGB1水平均高于对照组,差异有统计学意义(P0.05),在OA亚组中,KL2/3组滑膜和滑液HMGB1的水平均高于KL4组,差异有统计学意义(P0.05)。Spearman相关分析发现OA患者滑液HMGB1水平与滑膜炎、疼痛及日常活动评分呈显著正相关,与关节僵硬评分无显著相关。结论:膝OA患者关节功能可能主要受到关节局部病变的影响,而膝OA疼痛和僵硬的机制复杂,可能是局部和全身多种因素共同作用的结果;膝OA患者滑膜和滑液的HMGB1均过度表达,滑膜细胞表达的HMGB1可能作为一种新的促炎因子参与了膝OA疾病的发生发展,且主要在疾病的早中期发挥作用。滑液中的HMGB1的水平可能可以作为反映膝OA严重程度的生物学标志物。
[Abstract]:Objective: to evaluate the clinical symptoms, X-ray changes of knee joint and the relationship between them in patients with primary knee osteoarthritis (OAA), and to observe the expression and distribution of high mobility group protein B1HMGB1 on synovial cells of knee joint in patients with knee osteoarthritis. To explore the role of HMGB1 expressed by synovial cells in the development of knee OA disease, to analyze the relationship between the level of HMGB1 in synovial fluid and clinical symptoms and pathological changes of synovial membrane in patients with knee OA. To explore whether the level of HMGB1 in synovial fluid of knee OA patients can be used as a biological marker to reflect the severity of the disease and to provide a new method for early diagnosis of knee OA. Methods: 74 patients with primary knee OA who met the criteria were assessed with WOMAC scale before operation for knee joint pain, stiffness and daily activities, and KL grade was used to evaluate the severity of knee X-ray changes. According to KL grade, OA group was divided into two subgroups: KL2/3 group (moderate lesion group) and KL4 group (severe lesion group). The control group was 34 patients with cruciate ligament rupture or meniscus tear due to trauma in the same period. The control group had no history of OA and the X ray examination of knee joint was normal. In order to evaluate the severity of synovitis, the severity of synovitis was evaluated by grading synovial lesions from three aspects of lining layer proliferation, subintimal cell density and inflammatory cell infiltration under light microscope, and the distribution of HMGB1 in and out of synovial nucleus was observed by immunohistochemical technique. The percentage of synovial cells with positive HMGB1 staining was calculated, the relative level of HMGBl protein in synovial membrane was measured by Western blot, and the HMGB1 content in synovial fluid was measured by Elisa. The experimental results were statistically analyzed by SPSS21.0 software. Results the KL grade was significantly correlated with the total score of WOMAC and the score of daily activity, but not with the score of pain and stiffness. The results of HE staining showed that 30 cases of control group and 4 cases of OA group had normal synovium. Control group (n = 4) and OA group (n = 40) had mild synovitis and OA group (n = 30) with severe synovitis. The difference of synovitis score between the two groups was statistically significant (P 0.05). The expression of HMGB1 in synovial cells of the control group was mainly in the nucleus, while in the OA group, the expression of HMGB1 in the synovial cells was nuclear, extracellular and extracellular. Further counting showed that the proportion of synovial cells positive for HMGB1 expression in OA group was higher than that in control group (42.5 vs 39.7), the difference was statistically significant (P 0.05), and the proportion of synovial cells with positive expression of HMGB1 in nucleus in OA group was higher than that in control group (24.0% vs 5.7%), and the difference was statistically significant (P 0.05). And ELISA results showed that the levels of HMGB1 in synovium and synovial fluid in OA group were higher than those in control group. The level of HMGB1 in synovium and synovial fluid of KL 2 / 3 group was significantly higher than that of KL4 group. There was a significant positive correlation between HMGB1 level of synovial fluid and synovitis, pain and daily activity score in OA patients. There was no significant correlation with joint stiffness score. Conclusion: the joint function of patients with knee OA may be mainly affected by the local pathological changes of the joint, but the mechanism of pain and stiffness of knee OA is complicated, which may be the result of the joint action of various factors of the local and the whole body. HMGB1 in synovium and synovial fluid of knee OA patients were overexpressed. HMGB1 expressed by synovial cells may be a new pro-inflammatory factor involved in the occurrence and development of knee OA disease, and mainly play a role in the early and middle stage of the disease. The level of HMGB1 in synovial fluid may be used as a biomarker to reflect the severity of knee OA.
【学位授予单位】：新疆医科大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R684.3

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