沉默信息调节因子2在人骨关节炎软骨组织及细胞中表达的研究

发布时间：2018-03-12 16:22

本文选题：骨关节炎　切入点：沉默信息调节因子2　出处：《安徽医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：【研究目的】以人骨关节炎软骨标本研究对象,检测沉默信息调节因子2(SIRT2)在人软骨组织及细胞中的表达分布及表达情况,探讨SIRT2在骨关节炎(OA)与正常软骨组织及细胞中表达的差异性。【研究方法】选取临床中因膝骨关节炎而行全膝关节置换术患者的关节软骨标本20例,为OA组,按照2007年最新骨关节炎诊治指南的OA诊断标准,所有病人术前详细询问病史,无外伤、炎症、关节不稳定、慢性反复劳损或先天性疾病等。选取临床中因股骨颈骨折而行全髋关节置换术患者的关节软骨标本10例,为对照组,患者既往体健,否认既往存在关节疾病病史,此次股骨颈骨折根据患者病史及影像学检测排除OA。对取材关节软骨行免疫荧光染色检测SIRT2在两组软骨组织的表达分布及表达情况。在无菌条件下,分离软骨细胞并进行细胞传代培养,甲苯胺蓝染色进行细胞鉴定。采用Western blot法分别检测软骨组织及细胞中SIRT2的表达情况。【研究结果】免疫荧光染色结果显示SIRT2表达于软骨组织中,分布在细胞核与细胞质中,OA组SIRT2阳性细胞率较对照组明显降低(P0.01)。甲苯胺蓝染色后倒置显微镜下可见:大部分细胞呈现长梭形和多角形两种形态,细胞长满后排列紧密,细胞核为圆形或者椭圆形,位于胞体中心,细胞外基质均匀,被染成异染的蓝紫色,证实确为本实验所需关节软骨细胞。Western blot结果显示OA组软骨组织中SIRT2表达水平显著低于对照组(P0.01),软骨细胞中SIRT2的表达水平亦显著低于对照组(P0.01)。【结论】SIRT2在人类软骨组织中有表达,具体表达在细胞核与细胞质中,SIRT2表达降低可能导致骨关节炎的发生及进展。
[Abstract]:[objective] to detect the expression distribution and expression of silencing signaling factor 2sirt2 in human cartilage tissue and cells, using human osteoarthritis cartilage specimens. To investigate the difference of expression of SIRT2 in osteoarthritis and normal cartilage tissue and cell. [methods] 20 cases of osteoarthritis of knee joint underwent total knee arthroplasty were selected as OA group. In accordance with the OA diagnostic criteria of the latest guidelines for the diagnosis and treatment of osteoarthritis in 2007, all patients were asked about the history of the disease in detail before operation, without trauma, inflammation, instability of the joint. Chronic recurrent strain or congenital diseases. Ten articular cartilage specimens from patients undergoing total hip arthroplasty due to femoral neck fracture were selected as the control group. The patients were healthy and denied the existence of a history of joint disease. The femoral neck fracture was excluded according to the patient's history and imaging examination. The distribution and expression of SIRT2 in cartilage tissue of the two groups were detected by immunofluorescence staining. The chondrocytes were isolated and cultured. Toluidine blue staining was used to detect the expression of SIRT2 in chondrocytes and chondrocytes by Western blot method. [results] Immunofluorescence staining showed that SIRT2 was expressed in chondrocytes. The percentage of SIRT2 positive cells in OA group was significantly lower than that in control group (P 0.01). After toluidine blue staining, it was found that most of the cells showed long spindle shape and polygonal shape. The nucleus is round or elliptical, located at the center of the cell body, homogenous in the extracellular matrix, and dyed as a blue-purple heterochromatic. The results of Western blot showed that the expression of SIRT2 and SIRT2 in chondrocytes were significantly lower in OA group than in control group. [conclusion] SIRT2 was expressed in human chondrocytes. The decrease of SIRT2 expression in nucleus and cytoplasm may lead to the occurrence and progression of osteoarthritis.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R684.3

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