CAJ | 학술논문

[目的]探讨 C反应蛋白（CRP）和肿瘤坏死因子-α（TNF-α）对人外周血单核细胞分泌妊娠相关血浆蛋白-A （PAPP-A）的影响。[方法]采用密度梯度离心法分离及培养人外周血单核细胞，超敏ELISA法测定CRP或rhTNF-α刺激单核细胞后细胞上清液中PAPP-A的含量，Trans AMTM 技术检测CRP或rhTNF-α对单核细胞NF-κB活性的影响，并与空白对照组比较。同时观察给予核因子-κB （NF-κB）抑制剂 BAY11-70682预处理后，CRP 或 rhTNF-α对单核细胞合成分泌PAPP-A的影响。[结果]与空白对照组细胞上清液中PAPP-A含量（1．1±0．1） mIU/L相比，CRP（20 mg/L）刺激单核细胞2h后，PAPP-A含量开始显著增加（2．2±0．2）mIU/L，在24h达最高值（9．8±0．5）mIU/L（P ＜0．05），呈时间依赖性。给予rhTNF-α（100 ng/mL）刺激后，细胞上清液中PAPP-A含量在8 h达到高峰（14．5±1．1） mIU/L（ P ＜0．05），24h（11．7±0．9）mIU/L仍高于空白对照组（ P ＜0．05）。CRP和 rhTNF-α均呈剂量依赖性增加细胞上清液 PAPP-A 含量。CRP和rhTNF-α可增加单核细胞的磷酸化 NF-κB p65水平[分别为（0．88±0．06） mg/L、（1．06±0．12） mg/L]，显著高于对照组（0．35±0．04） mg/L（ P ＜0．05）。给予BAY11-70682预处理后，CRP和 rhTNF-α诱导单核细胞 NF-κB活性增加和分泌PAPP-A的作用受到抑制。[结论]CRP和TNF-α能促进人外周血单核细胞合成分泌PAPP-A ，其作用与NF-κB信号通路的激活有关。这可能是急性冠脉综合征患者血清PA PP-A升高的机制。
[목적]탐토 C반응단백（CRP）화종류배사인자-α（TNF-α）대인외주혈단핵세포분비임신상관혈장단백-A （PAPP-A）적영향。[방법]채용밀도제도리심법분리급배양인외주혈단핵세포，초민ELISA법측정CRP혹rhTNF-α자격단핵세포후세포상청액중PAPP-A적함량，Trans AMTM 기술검측CRP혹rhTNF-α대단핵세포NF-κB활성적영향，병여공백대조조비교。동시관찰급여핵인자-κB （NF-κB）억제제 BAY11-70682예처리후，CRP 혹 rhTNF-α대단핵세포합성분비PAPP-A적영향。[결과]여공백대조조세포상청액중PAPP-A함량（1．1±0．1） mIU/L상비，CRP（20 mg/L）자격단핵세포2h후，PAPP-A함량개시현저증가（2．2±0．2）mIU/L，재24h체최고치（9．8±0．5）mIU/L（P ＜0．05），정시간의뢰성。급여rhTNF-α（100 ng/mL）자격후，세포상청액중PAPP-A함량재8 h체도고봉（14．5±1．1） mIU/L（ P ＜0．05），24h（11．7±0．9）mIU/L잉고우공백대조조（ P ＜0．05）。CRP화 rhTNF-α균정제량의뢰성증가세포상청액 PAPP-A 함량。CRP화rhTNF-α가증가단핵세포적린산화 NF-κB p65수평[분별위（0．88±0．06） mg/L、（1．06±0．12） mg/L]，현저고우대조조（0．35±0．04） mg/L（ P ＜0．05）。급여BAY11-70682예처리후，CRP화 rhTNF-α유도단핵세포 NF-κB활성증가화분비PAPP-A적작용수도억제。[결론]CRP화TNF-α능촉진인외주혈단핵세포합성분비PAPP-A ，기작용여NF-κB신호통로적격활유관。저가능시급성관맥종합정환자혈청PA PP-A승고적궤제。
[Objective]To explore the effect of C-reactive protein(CRP) and tumor necrosis factor-α(TNF-α) on pregnancy associated plasma protein-A (PAPP-A ) secreted by human peripheral blood monocytes .[Methods] Human peripheral blood monocytes were isolated and cultured by using density gradient centrifugation .After monocytes were stimulated by CRP or rhTNF-α,supersensitive enzyme linked immunosorbent assay (ELISA) was used to detect the level of PAPP-A in supernatants . The effect of CRP or rhTNF-αon nuclear factor-κB(NF-κB) in monocytes was measured by TransAMTM technique ,and com-pared with control group .Meanwhile ,the effect of CRP or rhTNF-αon PAPP-A secreted by monocytes after NF-κB inhibitor BAY11-70682 treatment was observed .[Results]Compared with control group(1 .1 ± 0 .1mIU/L) ,PAPP-A level in culture su-pernatantsaftermonocytesstimulatedwithCRP(20mg/L)for2hbegantoincreasesignificantly(2.2±0.2mIU/L)andupto the peak value(9 .8 ± 0 .5mIU/L) after 24h in a time-dependent manner( P <0 .05) .After rhTNF-α(100ng/mL) stimulation , PAPP-A level increased up to the peak value(14 .5 ± 1 .1mIU/L) at 8h( P <0 .05) and the value(11 .7 ± 0 .9mIU/L) at 24h , which was still higher than that in control group ( P<0 .05) .CRP and rhTNF-αcould elevate PAPP level in supernatants in a dose-dependent manner .CRP and rhTNF-αcould increase phosphorylation NF-κB p65 level in monocytes[(0 .88 ± 0 .06mg/L) and (1 .06 ± 0 .12mg/L) ,respectively] ,which were higher than those in control group(0 .35 ± 0 .04mg/L)( P <0 .05) .After BAY11-70682 pretreatment ,the increasing of NF-κB activity and the secretion of PAPP-A in monocytes induced by CRP and rhTNF-αwere inhibited .[Conclusion]CRP and TNF-αcan promote the synthesis and secretion of PAPP-A in human peripheral blood monocytes ,which is related with the activation of NF-κB signal transduction and may be the mechanism of the elevation of serum PPP-A level in patients with acute coronary syndrome .