关节腔注射羧甲基壳聚糖对大鼠骨关节炎软骨核因子-κB活化及一氧化氮合酶表达的影响  

The effect of intra-articular injection of CM-chitosan on nuclear factor-κB activation and nitric oxide synthase expression in rat cartilage

作　　者：丁万军[1] 刘世清[2] 陶卫平[1] 贺斌[2] 

机构地区：[1]武汉大学人民医院肿瘤中心,430060 [2]武汉大学人民医院骨科,430060

出　　处：《中华风湿病学杂志》2013年第1期41-45,F0003,共6页Chinese Journal of Rheumatology

摘　　要：目的观察关节腔注射羧甲基壳聚糖（CMCTS）对骨关节炎模型中大鼠软骨细胞核因子-κB活化及诱导性一氧化氮合酶（iNOS）表达的影响。方法SD大鼠36只，采用随机数字表随机分假手术组、造模组、CMCTS组，每组12只，造模组与CMCTS组大鼠行切断膝关节内侧副韧带和切除部分内侧半月板的方式建立大鼠骨关节炎模型，术后5周CMCTS组关节腔内注射3％羧甲基壳聚糖0．15mg／kg，每周重复注射1次，术后11周处死动物，通过大体观察对各组关节软骨评分，通过甲苯胺蓝染色观察各组关节软骨的形态变化，通过免疫组织化学检测关节软骨细胞核因子-κB（P65）表达，通过蛋白印迹法检测关节软骨细胞I-κB蛋白表达和胞核P65蛋白表达，通过反转录-聚合酶链反应（RT—PCR）和蛋白印迹法分析软骨细胞iNOSmRNA和蛋白的表达。大体评分结果采用日检验，其余数据采用单因素方差分析。结果CMCTS组软骨退变大体评分明显优于造模组，CMCTS组关节软骨中核因子-κB表达（106±7）较B组（147±8）明显降低，CMCTS组明显抑制了关节软骨中I—κB的降解及软骨细胞iNOSmRNA和蛋白的表达，且CMCTS组抑制了关节软骨的退变。结论CMCTS可能通过抑制骨关节炎大鼠软骨细胞I-κB的降解而抑制核因子-κB信号通路的活化，从而下调骨关节炎大鼠软骨iNOS mRNA和蛋白的表达，保护骨关节软骨。Objective To observe the effect of intra-articular injection of CM-chitosan on nuclear factor κB （NF-κB） activation and nitric synthase expression in rat osteoarthritis cartilage, and to explore the destruction. Methods Thirty-six SD rats were randomly divided into three mechanism of inhibition of joint groups： A,B,C, 12 in each group. Group A was the sham group, group B, C rats had the medial collateral ligaments cut off and part of medial meniscus were removed to establish osteoarthristis model. Group C rats were injected with 3% carboxymethyl chitosan intra-articularly 0.15 mg/kg 5 weeks later, and then repeated injection every 1 week. Animals were sacrificed 11 weeks after surgery. The gross changes of cartilage and the expression of NF-KB （P65） were compared by immunohistochemistry, the protein expression of I-KB and P65 in nucleus were detected by Westem-bloting. Inducible nitric oxide synthase （iNOS） mRNA and protein expres-sion were analyzed by reverse transcription polymerase chain reaction （RT-PCR） and western blot. The general score of cartilage was analyzed by H test, the rest data was analyzed by one-way ANOVA. Results The cartilage degeneration scores pf group C （intra-articular injection of CM-chitosan group） were significantly less than those of group B. The protein expression of NF-κB of the articular cartilage in group C （106±7）was significantly lower than group B （147±8）, the I-κB degradation was inhibited significantly in group C , and the expression of iNOS mRNA and iNOS protein were reduced in OA articular cartilage of arthritis rat chondrocytes, therefore, it had protective effect on articular cartilage. Conclusion CMC may inhibit NF-κB signaling pathway by inhibiting the degradation of I-κB in cartilage, which reduces iNOS mRNA and protein expression in rat osteoarthritis cartilage, thereby protects rat osteoarthritis cartilage cells.

关 键 词：骨关节炎 软骨 细胞核因子类 一氧化氮合酶 羧甲基壳聚糖 

分 类 号：R563[医药卫生—呼吸系统]