透明质酸/壳聚糖微球对炎性软骨细胞基质金属蛋白酶及其抑制因子mRNA表达的影响  

Effects of hyaluronic acid and chitosan microspheres on mRNA expression of matrix metalloproteinases-1,-3,-13 and tissue inhibitor of metalloproteinases-1 in the inflammatory chondrocytes

作　　者：周庞虎[1] 马蓓蕾[2] 邱波[1] 彭昊[1] 刘世清[1] 

机构地区：[1]武汉大学人民医院骨关节外科,430060 [2]山东大学齐鲁医院检验科

出　　处：《中华实验外科杂志》2013年第11期2377-2379,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81071494）;湖北省自然科学基金资助项目（2011CHB021）;湖北省卫生厅青年科技人才资助项目（QJX2012-12）;武汉大学自主科研项目（302274670）

摘　　要：目的 观察透明质酸/壳聚糖微球（HA/CS）对重组大鼠白细胞介素-1β（rrIL-1β）诱导体外软骨细胞基质金属蛋白酶（MMP）及其抑制因子（TIMP） mRNA表达的影响,并探讨其作用机制,同时探讨透明质酸与壳聚糖保护软骨细胞作用的最佳比例.方法 体外分离、培养大鼠软骨细胞成功后,以10 μg/L IL-1β刺激模拟骨关节炎软骨细胞的炎性级联反应,培养24 h,分别加入透明质酸微球、壳聚糖微球、透明质酸/壳聚糖微球继续培养24 h,以逆转录-聚合酶链反应（RT-PCR）检测MMP-1、MMP-3、MMP-13及TIMP-1 mRNA的表达,通过Griess反应测定上清液中一氧化氮（NO）浓度.结果 透明质酸/壳聚糖微球可明显下调IL-1β诱导下软骨细胞MMP-1、MMP-3、MMP-13mRNA的表达（0.467±0.072、0.621±0.125、0.53±0.12）,但对TIMP-1 mRNA的表达（0.69±0.13）无明显影响.结论 透明质酸/壳聚糖微球能有效抑制IL-1β诱导的软骨细胞炎性级联反应,其保护作用可能与其抑制软骨细胞NO产生有关.Objective To investigate the effect of hyaluronic acid （HA）/chitosan （CS） microspheres on the expression of matrix metalloproteinases （MMPs）-1,-3 and-13 and tissue inhibitor of MMPs （TIMP-1） mRNA in chondrocytes exposed to interleukin （IL）-1 β in vitro,and study the mechanism,then explore the optimal ratio of HA to CS for protecting chondrocytes.Methods HA microspheres,CS nanopaticles and different ratio of HA/CS microspheres were added to culture medium for 24 h,respectively,after 10 μg/L IL-1 β was added to the culture medium of rat chondrocytes and co-cultured for 24 h.The expression levels of MMP-1,-3 and-13,and TIMP-1 mRNA were detected by using reverse transcriptasepolymerase chain reaction （RT-PCR）.The concentration of nitric oxide （NO） in the supernatants was measured by Griess reaction.Results HA/CS microspheres could significantly down-regulate the expression of MMP-1,-3,and-13 mRNA （0.467 ±0.072,0.621 ±0.125 and 0.53 ±0.12,respectively） of chondrocytes,while had little effects on the expression of TIMP-1 mRNA （0.69 ± 0.13）.Conclusion HA/CS could effectively interrupt the inflammatory cascade reaction caused by IL-1 β through inhibiting NO production.

关 键 词：透明质酸 壳聚糖 白细胞介素-1Β 基质金属蛋白酶 软骨细胞 

分 类 号：R684.3[医药卫生—骨科学]