中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2011年
7期
468-472
,共5页
韩潇%周道斌%许彩民%杨洋%段明辉%汪玄%张洁萍%赵永强%沈悌%武永吉
韓瀟%週道斌%許綵民%楊洋%段明輝%汪玄%張潔萍%趙永彊%瀋悌%武永吉
한소%주도빈%허채민%양양%단명휘%왕현%장길평%조영강%침제%무영길
贫血%红细胞生成素%hepcidin%白细胞介素6
貧血%紅細胞生成素%hepcidin%白細胞介素6
빈혈%홍세포생성소%hepcidin%백세포개소6
Anemia%Erythropoietin%Hepcidin%Interleukin-6
目的 探讨红细胞生成素(EPO)对单核细胞hepcidin的影响及其分子学机制.方法 采用实时定量PCR和Western blot方法检测hepcidin及信号分子C/EBPα、Smad1/5/8、磷酸化(p)-Smad1/5/8和p-STAT3的表达水平.采用IL-6或脂多糖(LPS)刺激人单核细胞系THP-1细胞,观察EPO对THP-1细胞hepcidin及信号分子表达的影响.加入EPO受体(EPOR)的抗体,观察其对EPO的拮抗作用以及对信号分子的影响.结果 EPO能抑制由20 ng/ml IL-6或1 μg/ml LPS诱导的THP1单核细胞hepcidin mRNA表达,作用呈时间和浓度限制性,EPO 2 U/ml(降低56%或64%)、作用6 h(降低53%或66%)抑制作用最明显.在20 ng/ml IL-6诱导作用下,EPO还可抑制hepcidin蛋白表达,EPO 2 U/ml作用6 h hepcidin蛋白表达明显降低.同时信号分子C/EBPα、p-Smad1/5/8和p-STAT3表达也受抑制,EPOR抗体能够拮抗EPO对hepcidin以及上述信号分子的抑制作用.结论 EPO可以抑制IL-6或LPS诱导的单核细胞hepcidin表达;在体外,EPO可能是通过下调C/EBPα、p-Smad1/5/8和p-STAT3进而抑制单核细胞hepcidin表达.
目的 探討紅細胞生成素(EPO)對單覈細胞hepcidin的影響及其分子學機製.方法 採用實時定量PCR和Western blot方法檢測hepcidin及信號分子C/EBPα、Smad1/5/8、燐痠化(p)-Smad1/5/8和p-STAT3的錶達水平.採用IL-6或脂多糖(LPS)刺激人單覈細胞繫THP-1細胞,觀察EPO對THP-1細胞hepcidin及信號分子錶達的影響.加入EPO受體(EPOR)的抗體,觀察其對EPO的拮抗作用以及對信號分子的影響.結果 EPO能抑製由20 ng/ml IL-6或1 μg/ml LPS誘導的THP1單覈細胞hepcidin mRNA錶達,作用呈時間和濃度限製性,EPO 2 U/ml(降低56%或64%)、作用6 h(降低53%或66%)抑製作用最明顯.在20 ng/ml IL-6誘導作用下,EPO還可抑製hepcidin蛋白錶達,EPO 2 U/ml作用6 h hepcidin蛋白錶達明顯降低.同時信號分子C/EBPα、p-Smad1/5/8和p-STAT3錶達也受抑製,EPOR抗體能夠拮抗EPO對hepcidin以及上述信號分子的抑製作用.結論 EPO可以抑製IL-6或LPS誘導的單覈細胞hepcidin錶達;在體外,EPO可能是通過下調C/EBPα、p-Smad1/5/8和p-STAT3進而抑製單覈細胞hepcidin錶達.
목적 탐토홍세포생성소(EPO)대단핵세포hepcidin적영향급기분자학궤제.방법 채용실시정량PCR화Western blot방법검측hepcidin급신호분자C/EBPα、Smad1/5/8、린산화(p)-Smad1/5/8화p-STAT3적표체수평.채용IL-6혹지다당(LPS)자격인단핵세포계THP-1세포,관찰EPO대THP-1세포hepcidin급신호분자표체적영향.가입EPO수체(EPOR)적항체,관찰기대EPO적길항작용이급대신호분자적영향.결과 EPO능억제유20 ng/ml IL-6혹1 μg/ml LPS유도적THP1단핵세포hepcidin mRNA표체,작용정시간화농도한제성,EPO 2 U/ml(강저56%혹64%)、작용6 h(강저53%혹66%)억제작용최명현.재20 ng/ml IL-6유도작용하,EPO환가억제hepcidin단백표체,EPO 2 U/ml작용6 h hepcidin단백표체명현강저.동시신호분자C/EBPα、p-Smad1/5/8화p-STAT3표체야수억제,EPOR항체능구길항EPO대hepcidin이급상술신호분자적억제작용.결론 EPO가이억제IL-6혹LPS유도적단핵세포hepcidin표체;재체외,EPO가능시통과하조C/EBPα、p-Smad1/5/8화p-STAT3진이억제단핵세포hepcidin표체.
Objective To investigate the in vitro effect of erythropoietin (EPO) on hepcidin of monocytes and its molecular mechanisms.Methods Hepcidin and signaling molecules including C/EBPα, Smad1/5/8,p-Smad1/5/8 and p-STAT3 were detected by real time PCR and Western blot.THP-1 monocytes were stimulated by interleukin-6 (IL-6 ) or lipopolysaccharide ( LPS).EPO receptor ( EPOR) antibody was added to observe its antagonistic effect on EPO and impact on the signaling proteins.Results EPO suppressed mRNA expression of THP-1 hepcidin of monocytes induced by 20 ng/ml IL-6 or 1 μg/ml LPS in both dose and time dependent manner.The most decrease of hepcidin expression was observed at 2 IU/ml EPO for 6 hours.EPO also down-regulated hepcidin protein induced by 20 ng/ml IL-6.At 2 IU/ml EPO for 6 hours hepcidin protein was down-reguleted, as was C/EBPα, p-Smad1/5/8 and p-STAT3.Antibody to EPOR antagonized the down-regulation of EPO on hepcidin and signaling proteins.Conclusion s Monoytes hepcidin can be reduced by EPO when stimulated by IL-6 or LPS.The mechanism of which may be at least in part, via suppression of C/EBPa, p-Smadl/5/8 and p-STAT3 signaling.
