中华老年医学杂志
中華老年醫學雜誌
중화노년의학잡지
Chinese Journal of Geriatrics
2012年
2期
140-143
,共4页
氨基糖类%骨关节炎%软骨细胞%一氧化氮
氨基糖類%骨關節炎%軟骨細胞%一氧化氮
안기당류%골관절염%연골세포%일양화담
Amino sugars%Osteoarthritis%Chondrocytes%Nitric oxide
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.
目的 研究硫痠氨基葡萄糖(GS)對白介素1β(IL-1β)誘導體外培養的人骨關節炎軟骨細胞(HOC)閤成一氧化氮(NO)的影響及其作用機製. 方法 取10例骨關節炎患者行全膝關節置換術的軟骨標本穫取軟骨細胞進行培養.在HOC培養液中加入IL 1β(5μg/L)和不同濃度的GS(0.2 mmol/L,2.0 mmol/L,20.0 mmol/L)作用24 h,酶聯免疫吸附(ELISA)測定檢測細胞上清中的NO的含量,反轉錄聚閤酶鏈反應(RT PCR)法和蛋白印跡法分彆檢測HOC中誘導型一氧化氮閤酶(iNOS)mRNA和蛋白的錶達. 結果 IL-1β刺激後HOC閤成NO增加,iNOS mRNA和蛋白錶達上調(t=-14.81,-45.38,均P＜0.01).2.0 mmol/L和20.0 mmol/L GS濃度依賴式抑製IL-1β誘導HOC閤成NO(F=12.43,P＜0.05),抑製IL-1β誘導HOC中iNOS mRNA(F=142.28,P＜0.05)和蛋白的上調(F=78.08,P＜0.01).單獨使用20.0 mmol/L的GS對HOC閤成NO無影響(t=-0.17,P＞0.05). 結論 GS通過下調HOC細胞內iNOS mRNA和蛋白的錶達從而抑製IL-1β誘導的NO閤成.
목적 연구류산안기포도당(GS)대백개소1β(IL-1β)유도체외배양적인골관절염연골세포(HOC)합성일양화담(NO)적영향급기작용궤제. 방법 취10례골관절염환자행전슬관절치환술적연골표본획취연골세포진행배양.재HOC배양액중가입IL 1β(5μg/L)화불동농도적GS(0.2 mmol/L,2.0 mmol/L,20.0 mmol/L)작용24 h,매련면역흡부(ELISA)측정검측세포상청중적NO적함량,반전록취합매련반응(RT PCR)법화단백인적법분별검측HOC중유도형일양화담합매(iNOS)mRNA화단백적표체. 결과 IL-1β자격후HOC합성NO증가,iNOS mRNA화단백표체상조(t=-14.81,-45.38,균P＜0.01).2.0 mmol/L화20.0 mmol/L GS농도의뢰식억제IL-1β유도HOC합성NO(F=12.43,P＜0.05),억제IL-1β유도HOC중iNOS mRNA(F=142.28,P＜0.05)화단백적상조(F=78.08,P＜0.01).단독사용20.0 mmol/L적GS대HOC합성NO무영향(t=-0.17,P＞0.05). 결론 GS통과하조HOC세포내iNOS mRNA화단백적표체종이억제IL-1β유도적NO합성.
Objective 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.
