CAJ | 학술논문

背景：植物凝集素可以刺激外周血单个核细胞分裂并引起免疫细胞的活化，是常用的细胞增殖模型。但是全血中的非外周血单个核细胞成分(血浆和无核细胞)在植物凝集素参与的体外培养过程中起到什么作用，以及内皮分泌因子的表达情况尚不明确。目的：在体外单独培养或全血培养外周血单个核细胞过程中，观察植物凝集素对其增殖和凋亡的影响，以及相关炎症因子以及内皮分泌因子的表达变化。方法：分离正常核型成人的外周血单个核细胞，应用无植物凝集素培养基和加入植物凝集素培养基分别进行单独培养，观察细胞形态学变化。收集全血培养或不同条件单独培养的外周血单个核细胞，提取mRNA，荧光定量RT-PCR检测细胞增殖、凋亡，以及炎症因子和内皮分泌因子的表达变化，并进行统计学分析。结果与结论：外周血单个核细胞单独培养和全血培养存在差异，植物凝集素会促进外周血单个核细胞 Ki67、增殖细胞核抗原、蛋白水解酶3、γ-干扰素、肿瘤坏死因子β和白细胞介素6的基因表达，抑制蛋白 C表达。提示植物凝集素促进体外培养外周血单个核细胞的增殖及凋亡，促使炎症因子表达上调，部分血液生物学相关的内皮分泌因子下调。
배경：식물응집소가이자격외주혈단개핵세포분렬병인기면역세포적활화，시상용적세포증식모형。단시전혈중적비외주혈단개핵세포성분(혈장화무핵세포)재식물응집소삼여적체외배양과정중기도십요작용，이급내피분비인자적표체정황상불명학。목적：재체외단독배양혹전혈배양외주혈단개핵세포과정중，관찰식물응집소대기증식화조망적영향，이급상관염증인자이급내피분비인자적표체변화。방법：분리정상핵형성인적외주혈단개핵세포，응용무식물응집소배양기화가입식물응집소배양기분별진행단독배양，관찰세포형태학변화。수집전혈배양혹불동조건단독배양적외주혈단개핵세포，제취mRNA，형광정량RT-PCR검측세포증식、조망，이급염증인자화내피분비인자적표체변화，병진행통계학분석。결과여결론：외주혈단개핵세포단독배양화전혈배양존재차이，식물응집소회촉진외주혈단개핵세포 Ki67、증식세포핵항원、단백수해매3、γ-간우소、종류배사인자β화백세포개소6적기인표체，억제단백 C표체。제시식물응집소촉진체외배양외주혈단개핵세포적증식급조망，촉사염증인자표체상조，부분혈액생물학상관적내피분비인자하조。
BACKGROUND:Phytohemagglutinin (PHA) can stimulate the peripheral blood mononuclear cells (PBMCs) into cellcycle, and cause their immune activation, which is a common immune proliferation model. However, the role of non-PBMC ingredient of peripheral blood is unclear, as wel as the expression of endothelial cells related cytokines. OBJECTIVE:To study the effect of whole blood culture and PBMCs alone culture with PHA on the PBMC proliferation and apoptosis, expression of inflammatory cytokine and endothelial cellsecreted cytokine markers. METHODS:Morphological changes of PBMCs separated from normal karyotype human peripheral blood individual y cultured with or without PHA were observed. The PBMCs were col ected by whole blood culture or PBMC separated culture. mRNA was extracted for the fluorescence quantitative RT-PCR, which was applied to detect the cellproliferation, apoptosis, and expression of inflammatory cytokine and endothelial cellsecreted cytokines. The statistic analysis was used for the significance explication. RESULTS AND CONCLUSION:PBMCs alone cultured ere different from those undergoing whole blood culture. The PHA could up-regulate the gene expression of Ki67, proliferating cellnuclear antigen, Caspase 3, interferon-γ, tumor necrosis factor-βand interleukin-6, but down-regulate Protein C. This indicted that PHA could promote the proliferation and apoptosis of PBMCs and up-regulate the expression of inflammatory cytokines, but down-regulate the expression of endothelial cells secreted coagulation cytokines.