透明质酸/壳聚糖微球对骨关节炎软骨细胞iNOS活性和表达的影响

Effects of Hyaluronic Acid and Chitosan Microspheres on Inducible Oxide Synthase Activity and Expression in Osteoarthritis Chondrocytes

目的:研究透明质酸/壳聚糖微球对重组大鼠白细胞介素-1β(rrIL-1β)诱导体外骨关节炎软骨细胞诱导型一氧化氮合成酶(iNOS)活性和表达的影响,并初步探讨其作用机制。方法:体外分离、培养大鼠软骨细胞成功后,以10ng/L IL-1β刺激模拟软骨细胞炎性级联反应,培养1h后分别加入透明质酸微球、壳聚糖微球和透明质酸/壳聚糖微球,继续培养24h。通过RT-PCR检测iNOS mRNA的表达,利用Griess反应测定上清液中一氧化氮(NO)的浓度,通过iNOS催化L-精氨酸和分子氧反应生成NO推算iNOS的活性。结果:透明质酸/壳聚糖微球能有效抑制IL-1β刺激诱导的软骨细胞iNOS的活性和NO的产量;RT-PCR检测显示透明质酸/壳聚糖微球能抑制iNOS mRNA的表达。结论:透明质酸/壳聚糖微球能通过抑制骨关节炎软骨细胞iNOS活性及蛋白表达来减少骨关节炎软骨细胞中NO的产生。

Objective： To investigate the effecs of hyaluronic acid（HA） and chitosan（CS） microspheres on inducible oxide synthase activity and expression in chondrocytes exposured to rrIL-1β in vitro. Methods： HA（microspheres）, CS（microspheres） and HA/CS（microspheres） were added to cul- ture medium for 24 hours, respectively, after 10 ng/ml IL--1β were added to the culture medium of rat chondrocytes and co-cultured for 1 hours. Then, we examined the expressions of inducible nitric oxide synthase （iNOS） mRNA and activity of iNOS by RT-PCR. The concentration of ni- tric oxide （NO） in the supernatants were measured by Griess reaction. Results： HA and CS（mi- crospheres） could inhibit the up-regulated iNOS activity and NO production induced by IL-1β ef- fectively. RT-PCR results showed that the expression of iNOS mRNA was inhibited by HA and CS（microspheres）. Conclusion： HA and CS（microspheres） could decrease the NO production in osteoarthritis chondrocytes by inhibiting the activity and mRNA expression of iNOS.