摘要目的 研究硫酸氨基葡萄糖(GS)对白介素1β(IL-1β)诱导体外培养的人骨关节炎软骨细胞(HOC)合成一氧化氮(NO)的影响及其作用机制. 方法 取10例骨关节炎患者行全膝关节置换术的软骨标本获取软骨细胞进行培养.在HOC培养液中加入IL 1β(5μg/L)和不同浓度的GS(0.2 mmol/L,2.0 mmol/L,20.0 mmol/L)作用24 h,酶联免疫吸附(ELISA)测定检测细胞上清中的NO的含量,反转录聚合酶链反应(RT PCR)法和蛋白印迹法分别检测HOC中诱导型一氧化氮合酶(iNOS)mRNA和蛋白的表达. 结果 IL-1β刺激后HOC合成NO增加,iNOS mRNA和蛋白表达上调(t=-14.81,-45.38,均P＜0.01).2.0 mmol/L和20.0 mmol/L GS浓度依赖式抑制IL-1β诱导HOC合成NO(F=12.43,P＜0.05),抑制IL-1β诱导HOC中iNOS mRNA(F=142.28,P＜0.05)和蛋白的上调(F=78.08,P＜0.01).单独使用20.0 mmol/L的GS对HOC合成NO无影响(t=-0.17,P＞0.05). 结论 GS通过下调HOC细胞内iNOS mRNA和蛋白的表达从而抑制IL-1β诱导的NO合成.

abstractsObjective To study the effects of glucosamine sulfate on nitric oxide(NO)production induced by interleukin(IL)-1β in human osteoarthritis chondrocytes(HOC),and explore the possible mechanism.Methods Chondrocytes were harvested from 10 osteoarthritis patients undergoing total knee replacement(TKR)operation.Human recombinant IL-1β(5 μg/L)and glucosamine sulfate GS in different concentrations(0.2 mmol/L,2.0 mmol/L,20.0 mmol/L)were administrated into cell culture medium for 24 h.The content of NO was detected by enzyme-linked immunosorbent assay(ELISA).The mRNA and protein expression of inductive nitric oxide synthetase(iNOS)were measured by RT-PCR and Western blot,respectively.Results Stimulation of HOC with IL-1β enhanced production of NO and expressions of iNOS mRNA and protein(t=-14.81,-45.38,all P＜0.01).Pretreatment with 2.0 and 20.0 mmol/L GS showed a dose-dependent inhibition of IL-1β induced NO production(F=12.43,P＜0.05)and the expression levels ofiNOSmRNA(F=142.28,P＜0.05)and protein(F=78.08,P＜0.01).20.0 mmol/L GS alone did not influence NO production(t =-0.17,P＞ 0.05).Conclusions GS may inhibit the synthesis of NO induced by IL-1β in HOC through down-regulate mRNA and protein expressions of iNOS.