中国老年学杂志
中國老年學雜誌
중국노년학잡지
CHINESE JOURNAL OF GERONTOLOGY
2014年
20期
5762-5765
,共4页
妊娠相关血浆蛋白-A%单核细胞%肿瘤坏死因子-α%核因子-κB
妊娠相關血漿蛋白-A%單覈細胞%腫瘤壞死因子-α%覈因子-κB
임신상관혈장단백-A%단핵세포%종류배사인자-α%핵인자-κB
Pregnancy associated plasma protein-A%Monocytes%Tumor necrosis factor-α%Nuclear factor-κB
目的：探讨肿瘤坏死因子-α（TNF-α）对人外周血单核细胞妊娠相关血浆蛋白（PAPP）-A蛋白和 mRNA表达的影响及其机制。方法采用密度梯度离心法分离及培养人外周血单核细胞，采用Western印迹法和实时荧光定量PCR法观察TNF-α诱导单核细胞PAPP-A蛋白和mRNA表达的时间及剂量效应。采用TransAMTM技术检测TNF-α对单核细胞 NF-κB活性的影响。此外观察给予核转录因子κB（NF-κB）抑制剂 BAY11-70682预处理后，TNF-α对单核细胞PAPP-A表达的影响。结果 TNF-α（100 ng/ml）刺激单核细胞后，PAPP-A的蛋白和mRNA表达分别在8 h和2 h达高峰，且呈剂量依赖性诱导单核细胞的PAPP-A表达。 TNF-α可显著增加单核细胞的磷酸化NF-κB p65水平，在给与BAY11-70682预处理后，TNF-α诱导PAPP-A表达的作用受到抑制。结论促炎因子TNF-α通过激活NF-κB途径上调单核细胞PAPP-A的蛋白和基因表达，这可能是急性冠脉综合征（ ACS）患者血清PAPP-A升高的机制之一。
目的：探討腫瘤壞死因子-α（TNF-α）對人外週血單覈細胞妊娠相關血漿蛋白（PAPP）-A蛋白和 mRNA錶達的影響及其機製。方法採用密度梯度離心法分離及培養人外週血單覈細胞，採用Western印跡法和實時熒光定量PCR法觀察TNF-α誘導單覈細胞PAPP-A蛋白和mRNA錶達的時間及劑量效應。採用TransAMTM技術檢測TNF-α對單覈細胞 NF-κB活性的影響。此外觀察給予覈轉錄因子κB（NF-κB）抑製劑 BAY11-70682預處理後，TNF-α對單覈細胞PAPP-A錶達的影響。結果 TNF-α（100 ng/ml）刺激單覈細胞後，PAPP-A的蛋白和mRNA錶達分彆在8 h和2 h達高峰，且呈劑量依賴性誘導單覈細胞的PAPP-A錶達。 TNF-α可顯著增加單覈細胞的燐痠化NF-κB p65水平，在給與BAY11-70682預處理後，TNF-α誘導PAPP-A錶達的作用受到抑製。結論促炎因子TNF-α通過激活NF-κB途徑上調單覈細胞PAPP-A的蛋白和基因錶達，這可能是急性冠脈綜閤徵（ ACS）患者血清PAPP-A升高的機製之一。
목적：탐토종류배사인자-α（TNF-α）대인외주혈단핵세포임신상관혈장단백（PAPP）-A단백화 mRNA표체적영향급기궤제。방법채용밀도제도리심법분리급배양인외주혈단핵세포，채용Western인적법화실시형광정량PCR법관찰TNF-α유도단핵세포PAPP-A단백화mRNA표체적시간급제량효응。채용TransAMTM기술검측TNF-α대단핵세포 NF-κB활성적영향。차외관찰급여핵전록인자κB（NF-κB）억제제 BAY11-70682예처리후，TNF-α대단핵세포PAPP-A표체적영향。결과 TNF-α（100 ng/ml）자격단핵세포후，PAPP-A적단백화mRNA표체분별재8 h화2 h체고봉，차정제량의뢰성유도단핵세포적PAPP-A표체。 TNF-α가현저증가단핵세포적린산화NF-κB p65수평，재급여BAY11-70682예처리후，TNF-α유도PAPP-A표체적작용수도억제。결론촉염인자TNF-α통과격활NF-κB도경상조단핵세포PAPP-A적단백화기인표체，저가능시급성관맥종합정（ ACS）환자혈청PAPP-A승고적궤제지일。
Objective To observe the effects of tumor necrosis factor-α( TNF-α) on pregnancy associated plasma protein-A ( PAPP-A) protein and mRNA expression in human peripheral blood monocytes and the underlying mechanismMethods Human monocytes were obtained from healthy volunteers by Ficoll-plaque centrifugation.Expression of PAPP-A protein and mRNA in monocytes stimulated with TNF-αwere detected by Western blot and quantitative real-time PCR.The level of nuclear factor-κB p65 (NF-κB p65) was measured by Trans AMTM technique.Moreover, the effect of TNF-αon PAPP-A expression after pretreatment with NF-κB inhibitor (BAY11-70682) was analyzed.Results PAPP-A protein and mRNA levels in monocytes stimulated by TNF-α(100 ng/ml) increased at 2 hours and reached peak at 8 hours and 2 hours respectively, and the induction effects of TNF-αwas dose-dependent.TNF-αcould significantly increase the level of NF-κB p65 in monocytes.Furthermore, BAY11-7082 completely inhibited up-regulation of PAPP-A protein and mRNA expression induced by TNF-α, indicating that NF-κB activation is necessary.Conclusions TNF-αcan up-regulate protein and gene expression of PAPP-A in human peripheral blood monocytes by activating NF-κB pathway, which may result in increase of serum PAPP-A level in patients with acute coronary syndromes.
