一氧化氮合酶抑制剂对白细胞介素1β诱导软骨细胞增殖和基质金属蛋白酶表达的影响  

Effects of nitricoxide synthase inhibitor on interleukin-1 beta-induced chondrocyte proliferation and matrix metalloproteinase expression

作　　者：梁鹏展[1] 江建明[1] 刘传芳[1] 孙玮[2] 余磊[3] 邱小忠[3] 

机构地区：[1]南方医科大学附属南方医院脊柱骨病外科,广东省广州市510515 [2]深圳市第二人民医院脊柱外科,广东省深圳市518035 [3]南方医科大学解剖教研室,广东省组织构建与检测重点实验室,广东省广州市510515

出　　处：《中国组织工程研究与临床康复》2008年第15期2820-2824,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30571890)~~

摘　　要：目的:研究表明,白细胞介素1β对鼠肾细胞、心肌细胞和神经上皮细胞发挥抗增殖作用,但其是否可抑制人的软骨细胞增殖?这种抗增殖作用是否可被一氧化氮合酶抑制剂抑制尚不清楚。实验拟验证一氧化氮合酶抑制剂对人骨关节炎软骨细胞增殖及基质金属蛋白酶的影响。方法:选择2006-05/12在南方医科大学附属南方医院脊柱骨病科住院的膝骨关节炎患者8例。所有患者均符合1986年美国风湿病学会关于骨关节炎的临床诊断标准或临床及放射学诊断标准,排除其他疾病(创伤性、风湿性及感染性)所导致的骨关节炎。所有患者均在术前告知,并签有试验知情同意书。分离培养关节软骨细胞,将不同浓度(0,1,10,100μg/L,其中0μg/L作为对照)白细胞介素1β作用于骨关节炎软骨细胞24,48,72h,一氧化氮合酶抑制剂氨基胍作为干扰因素,与白细胞介素1β共同作用72h,收集细胞上清液,一氧化氮的表达量通过一氧化氮试剂盒检测,并作为检测一氧化氮合酶抑制剂活性的手段;四甲基偶氮唑盐法检测软骨细胞的增殖;xMAP技术检测软骨细胞基质金属蛋白酶1,基质金属蛋白酶3和基质金属蛋白酶9的表达。结果:①不同时间点1,10,100μg/L白细胞介素1β组作用后的吸光度值与对照组相比,差异显著(P<0.05,P<0.01);相同浓度白细胞介素1β在24,72h的吸光度值相比较,差异显著(P<0.01)。②白细胞介素1β和氨基胍(100μmol/L)共同作用软骨细胞72h后,各浓度组吸光度与白细胞介素1β组相比,差异显著(P<0.01)。③白细胞介素1β可促进软骨细胞一氧化氮的表达,并呈剂量依赖关系。④白细胞介素1β促进基质金属蛋白酶1和基质金属蛋白酶3的表达,与对照组相比,差异有显著性意义(P<0.01),而软骨细胞基质金属蛋白酶9的表达在本实验中没有检测到。结论:白细胞介素1β可抑制软骨细胞增殖,诱导基质金属蛋白酶表达,这些影响�AIM： Studies have demonstrated that interleukin-1β （IL-1β ） can inhibit the proliferation of rat renal cells, myocardial cells, and neuroepithelial cells. But whether it can inhibit human chondrocyte proliferation, and whether this inhibitory effect can be suppressed by nitricoxide synthase （NOS） are still uncertain. This study explored the effects of NOS inhibitor on chondrocyte proliferation and matrix metalloproteinase （MMP） in patients with osteoarthritis. METHODS： Eight inpatients with osteoarthritis were selected from Department of Orthopaedics and Spine Surgery, Nanfang Hospital, Southern Medical University from May to December 2006. All subjects fulfilled the diagnosis criteria for osteoarthritis according to the diagnostic standards by American College of Rheumatology or the clinical and radiological diagnostic standards. Osteoarthritis induced by traumatic, rheumatic or infectious diseases was excluded. The informed consent was obtained from all subjects. After cell isolation and culture, chondrocytes were stimulated with different concentrations （0, 1, 10, 100 μg/L, 0 as control） IL- 1β for 24, 48, and 72 hours, separately and treated with aminoguanidine （AG）, a kind of NOS inhibitor, for 72 hours. Nitric oxide （NO） was detected using NO kit to identify the activity of NOS inhibitor. Cell proliferation was examined by MTT array, and MMP-1, MMP-3, and MMP-9 production in supernatants were measured by technique of flexible Multi-Analyte Profiling （xMAP）. RESULTS： （1）At different time points, there were significant differences in absorbance values between IL-1β groups （1, 10, 100 μg/L） and control group （P 〈 0.05, P 〈 0.01）; the absorbance values of the same concentration of IL-1β were significantly different at 24 and 72 hours of stimulation （P 〈 0.01）. （2）The absorbance values of chondrocytes treated with IL-1β and AG for 72 hours were statistically significant compared with cells only stimulated with IL-1β （P 〈 0.01）. （3）IL-

关 键 词：软骨细胞 骨关节炎 白细胞介素1Β 一氧化氮合酶抑制剂 一氧化氮 液相蛋白芯片 

分 类 号：R684.3[医药卫生—骨科学]