CAJ | 학술논문

目的：通过检测ASK1和PRKCD在单核细胞分化过程中的表达变化，探索其在门静脉高压症(PH)脾亢脾脏巨噬细胞（MΦ）功能变化中所起的作用。方法采用佛波酯诱导U937分化成为巨噬细胞样细胞，q-PCR检测不同分化阶段ASK1和PRKCD mRNA的表达，Western Blot检测诱导前后ASK1和PRKCD蛋白表达水平变化，ELISA法检测细胞分化过程中与吞噬功能相关的细胞因子IL-10和TNF-α的分泌情况，鸡红细胞吞噬试验鉴定诱导后细胞功能。结果随着PMA诱导U937细胞时间延长，ASK1和PRKCD基因表达水平下降，TNF-α的分泌量逐渐增加，而IL-10在诱导后24 h达到最大分泌量，随后又呈下降趋势；Western Blot结果显示，ASK1及p-ASK1蛋白表达水平较诱导前均显著上升，PRKCD及p-PRKCD蛋白表达水平显著下降；吞噬实验结果显示，诱导后的细胞具有一定的吞噬功能。结论在PMA诱导的U937细胞向单核细胞分化过程中，ASK1蛋白表达上调，PRKCD蛋白表达下调，与脾亢脾Mφ一致，并导致Mφ活性增强。
목적：통과검측ASK1화PRKCD재단핵세포분화과정중적표체변화，탐색기재문정맥고압증(PH)비항비장거서세포（MΦ）공능변화중소기적작용。방법채용불파지유도U937분화성위거서세포양세포，q-PCR검측불동분화계단ASK1화PRKCD mRNA적표체，Western Blot검측유도전후ASK1화PRKCD단백표체수평변화，ELISA법검측세포분화과정중여탄서공능상관적세포인자IL-10화TNF-α적분비정황，계홍세포탄서시험감정유도후세포공능。결과수착PMA유도U937세포시간연장，ASK1화PRKCD기인표체수평하강，TNF-α적분비량축점증가，이IL-10재유도후24 h체도최대분비량，수후우정하강추세；Western Blot결과현시，ASK1급p-ASK1단백표체수평교유도전균현저상승，PRKCD급p-PRKCD단백표체수평현저하강；탄서실험결과현시，유도후적세포구유일정적탄서공능。결론재PMA유도적U937세포향단핵세포분화과정중，ASK1단백표체상조，PRKCD단백표체하조，여비항비Mφ일치，병도치Mφ활성증강。
Objective To investigate the expression of ASK1 and PRKCD in the process of monocyte differentiation, and explore their role in functional changes of hypersplenism spleen macrophages (MΦ) in portal hypertension (PH). Methods U937 cells were stimulated to differentiate into monocyte/macrophage-like cells by cultivation in PMA and the mRNA expressions of ASK1 and PRKCD were detected by q-PCR and the changes of protein expression were identified by western blot analysis. The secretion of phagocytose related cytokines such as IL-10 and TNF-a were tested by ELISA, and the function of the macrophage-like cells were studied by chicken red blood cell phagocytose test. Results The expressions of PRKCD and ASK1 mRNA were gradually decreased along with the cell differentiation, while the secretion of TNF-αwas increased, IL-10 secretion reached a maximum at 24 h after PAM stimulation, and then gradually fell. The expression of ASK1 and p-ASK1 were rapidly increased compared with the non-stimulated U937 cells, while the expression of PRKCD and p-PRKCD were sightly declined. The phagocytose test show that U937 cells induced with PMA were able to swallow the chicken red blood cell.Conclusion Up-regulated protein expression of ASK1 and p-ASK1 and down-regulated protein expression of PRKCD and p-PRKCD in the process of PMA induced monocyte differentiation, are consist with the expression changes of splenic macrophage phagocytosis in hypersplenism, which leads to increased activity of MΦ.