水通道蛋白3在膝骨关节炎患者退变关节软骨细胞中的表达及意义

Expression and its significance of aquaporin 3 in degenertive articular chondrocytes of patients with knee osteoarthritis

目的测定水通道蛋白3(aquaporin 3,AQP3)在膝骨关节炎患者退变关节软骨细胞中的表达,探讨AQP3在膝骨关节炎退变关节软骨细胞中的作用及意义。方法选取自2009-01—2012-01芜湖市第一人民医院收治的36例因膝骨关节炎、外伤高位截肢、暴力骨折而进行全膝关节置换术、截肢术、切开复位内固定术并取出的关节软骨。实验组为20例膝骨关节炎,关节软骨为Outerbridge标准Ⅲ级;对照组为16例外伤高位截肢、暴力骨折,关节软骨为Outerbridge标准Ⅰ级。将术中取出的关节软骨采用酶消化法分离及培养出原代关节软骨细胞,倒置相差显微镜及HE染色观察软骨细胞形态,甲苯胺蓝染色及Real time RT-PCR对终板软骨细胞标志性基因Ⅱ型胶原及蛋白多糖进行测定、鉴定。Real time RT-PCR、Western blot法检测AQP3基因及蛋白的表达变化。结果对照组关节软骨细胞以多角形为主;实验组关节软骨细胞呈梭形。蛋白多糖、Ⅱ型胶原、AQP3基因在实验组中的表达较对照组低;AQP3蛋白在实验组中表达较对照组下调。结论膝骨关节炎患者关节软骨退变后,AQP3基因表达明显下降,通过调控关节软骨细胞中AQP3表达可能会改善关节软骨的退变。

Objective To observe the change of aquaporin 3 expression in articular chondrocytes of patients with knee osteoarthritis, and investigate the function and significance of aquaporin 3 in the degenerative chondrocytes of patients with knee osteoarthritis. Methods Articular cartilage was obtained from 36 inpatients who were used by total knee replacement,amputation, and open reduction and internal fixation for knee osteoarthritis, high amputation of trauma, traumatic fracture in the First People's Hospital of Wuhu from January 2009 to January 2012. These patients were divided into control group(n =16)with traumatic fracture or high amputation of trauma, and their articular cartilage for Outerbridge grade standard III, and experimental group(n =20) with osteoarthritis and their articular cartilage for Outerbridge grade standard Ⅰ. Articular chondrocytes were isolated by enzyme digestion and cultured in vitro. The morphological appearances and biological characteristics of articular chondrocytes were observed by inverted phase contrast microscope, HE staining, teluidine blue staining and real time RT-PCR respectively. Real time RT-PCR was used to detect the mRNA expresson of AQP3, aggrecan and type Ⅱ collagen. Western blot was used to analyze the change of AQP3 protein expression. Results The shape of articular chondrocytes in control group was mainly polygonal, the experimental group was fusiform; the mRNA expression of AQP3, aggrecan and typeⅡ collagen in the experimental group was much lower than the control group; compared with control group, the expression of AQP3 protein in the experimental group was decreased. Conclusion AQP3 gene expression was significantly decreased after articular chondrocytes degeneration in the knee osteoarthritis, regulating the expression of AQP3in knee articular chondrocytes which may improve the degeneration of articular cartilage.