CAJ | 학술논문

目的：观察髓系细胞触发受体-1（triggering receptor expressed on myeloid cells-1， TREM-1）在脐带血白细胞膜表达和mRNA转录水平，并分析其促进炎症细胞因子分泌的功能。方法2013年9月至2014年3月，分别收集20例健康足月新生儿出生时的脐带血和20例健康成年人的外周血，采用流式细胞术检测中性粒细胞和单核细胞TREM-1平均荧光强度和TREM-1阳性比例，实时荧光定量逆转录-聚合酶链反应技术检测白细胞TREM-1 mRNA。分别用脂多糖或LP17+脂多糖刺激脐带血白细胞，用酶联免疫吸附法检测上清液中可溶性TREM-1、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素（interleukin，IL）-6、IL-8浓度，并与未经刺激者（空白对照）作比较。采用成组t检验、方差分析、q检验和Pearson相关分析进行统计学分析。结果新生儿脐带血白细胞TREM-1的平均荧光强度与成年人比较，差异无统计学意义（P＞0.05）。新生儿中性粒细胞TREM-1阳性百分率为（82.3±7.1）%，低于成年人的（98.6±4.8）%，差异有统计学意义（t=8.512，P＜0.01）。新生儿TREM-1 mRNA水平为1.16±0.13，低于成年人的1.63±0.24，差异有统计学意义（t=7.714，P＜0.01）。新生儿脐带血未经脂多糖刺激，24 h后上清液sTREM-1浓度为（34.6±6.1）pg/ml；经脂多糖刺激24 h后，上清液sTREM-1浓度上升至（156.7±36.3）pg/ml，差异有统计学意义（t=13.623，P＜0.01）。LP17联合脂多糖处理后的脐带血IL-6、TNF-α、IL-8浓度明显低于仅经脂多糖处理的脐带血（P值均＜0.05）。新生儿脐带血经脂多糖刺激24 h后，上清液中sTREM-1浓度与TNF-α、IL-6、IL-8浓度呈正相关（r值分别0.519、0.507和0.538，P值分别为0.019、0.022和0.014）。结论新生儿脐带血单核细胞TREM-1发育情况与成年人一致，中性粒细胞TREM-1阳性比例低于成年人。阻断脐带血白细胞TREM-1可部分减少白细胞炎症细胞因子的分泌。目的：觀察髓繫細胞觸髮受體-1（triggering receptor expressed on myeloid cells-1， TREM-1）在臍帶血白細胞膜錶達和mRNA轉錄水平，併分析其促進炎癥細胞因子分泌的功能。方法2013年9月至2014年3月，分彆收集20例健康足月新生兒齣生時的臍帶血和20例健康成年人的外週血，採用流式細胞術檢測中性粒細胞和單覈細胞TREM-1平均熒光彊度和TREM-1暘性比例，實時熒光定量逆轉錄-聚閤酶鏈反應技術檢測白細胞TREM-1 mRNA。分彆用脂多糖或LP17+脂多糖刺激臍帶血白細胞，用酶聯免疫吸附法檢測上清液中可溶性TREM-1、腫瘤壞死因子-α（tumor necrosis factor-α，TNF-α）、白細胞介素（interleukin，IL）-6、IL-8濃度，併與未經刺激者（空白對照）作比較。採用成組t檢驗、方差分析、q檢驗和Pearson相關分析進行統計學分析。結果新生兒臍帶血白細胞TREM-1的平均熒光彊度與成年人比較，差異無統計學意義（P＞0.05）。新生兒中性粒細胞TREM-1暘性百分率為（82.3±7.1）%，低于成年人的（98.6±4.8）%，差異有統計學意義（t=8.512，P＜0.01）。新生兒TREM-1 mRNA水平為1.16±0.13，低于成年人的1.63±0.24，差異有統計學意義（t=7.714，P＜0.01）。新生兒臍帶血未經脂多糖刺激，24 h後上清液sTREM-1濃度為（34.6±6.1）pg/ml；經脂多糖刺激24 h後，上清液sTREM-1濃度上升至（156.7±36.3）pg/ml，差異有統計學意義（t=13.623，P＜0.01）。LP17聯閤脂多糖處理後的臍帶血IL-6、TNF-α、IL-8濃度明顯低于僅經脂多糖處理的臍帶血（P值均＜0.05）。新生兒臍帶血經脂多糖刺激24 h後，上清液中sTREM-1濃度與TNF-α、IL-6、IL-8濃度呈正相關（r值分彆0.519、0.507和0.538，P值分彆為0.019、0.022和0.014）。結論新生兒臍帶血單覈細胞TREM-1髮育情況與成年人一緻，中性粒細胞TREM-1暘性比例低于成年人。阻斷臍帶血白細胞TREM-1可部分減少白細胞炎癥細胞因子的分泌。
목적：관찰수계세포촉발수체-1（triggering receptor expressed on myeloid cells-1， TREM-1）재제대혈백세포막표체화mRNA전록수평，병분석기촉진염증세포인자분비적공능。방법2013년9월지2014년3월，분별수집20례건강족월신생인출생시적제대혈화20례건강성년인적외주혈，채용류식세포술검측중성립세포화단핵세포TREM-1평균형광강도화TREM-1양성비례，실시형광정량역전록-취합매련반응기술검측백세포TREM-1 mRNA。분별용지다당혹LP17+지다당자격제대혈백세포，용매련면역흡부법검측상청액중가용성TREM-1、종류배사인자-α（tumor necrosis factor-α，TNF-α）、백세포개소（interleukin，IL）-6、IL-8농도，병여미경자격자（공백대조）작비교。채용성조t검험、방차분석、q검험화Pearson상관분석진행통계학분석。결과신생인제대혈백세포TREM-1적평균형광강도여성년인비교，차이무통계학의의（P＞0.05）。신생인중성립세포TREM-1양성백분솔위（82.3±7.1）%，저우성년인적（98.6±4.8）%，차이유통계학의의（t=8.512，P＜0.01）。신생인TREM-1 mRNA수평위1.16±0.13，저우성년인적1.63±0.24，차이유통계학의의（t=7.714，P＜0.01）。신생인제대혈미경지다당자격，24 h후상청액sTREM-1농도위（34.6±6.1）pg/ml；경지다당자격24 h후，상청액sTREM-1농도상승지（156.7±36.3）pg/ml，차이유통계학의의（t=13.623，P＜0.01）。LP17연합지다당처리후적제대혈IL-6、TNF-α、IL-8농도명현저우부경지다당처리적제대혈（P치균＜0.05）。신생인제대혈경지다당자격24 h후，상청액중sTREM-1농도여TNF-α、IL-6、IL-8농도정정상관（r치분별0.519、0.507화0.538，P치분별위0.019、0.022화0.014）。결론신생인제대혈단핵세포TREM-1발육정황여성년인일치，중성립세포TREM-1양성비례저우성년인。조단제대혈백세포TREM-1가부분감소백세포염증세포인자적분비。
Objective To investigate the triggering receptor expressed on myeloid cells-1(TREM-1) of cord blood leukocytes in neonates and the transcription level of mRNA, and analyze its promoting function of inflammatory cytokine secretion. Methods During the period from September 2013 to March 2014, cord blood was collected from 20 term neonates at the time of birth, and peripheral blood was collected from 20 healthy adults. The expression of TREM-1 and TREM-1 mRNA on leukocytes was observed using flow cytometry and real-time reverse transeription-polymerase chain reaction, respectively. After the whole cord blood was stimulated by lipopolysaccharide (LPS) or LP17 plus LPS, the contents of interleukin (IL)-6, tumor necrosis factor-α (TNF-α), IL-8 and soluble TREM-1 (sTREM-1) in the supernatant were analyzed by enzyme linked immunosorbent assay. The statistical significance was determined using the one-way ANOVA test, t test, q test and Pearson correlation coefficient. Results The mean fluorescence intensity of TREM-1 on leukocytes of newborns was not different compared with healthy adults (P>0.05), while the percentage of TREM-1 positive on polymorphonuclear cells was lower than that of healthy adults [(82.3±7.1)% vs (98.6±4.8)%, P<0.05]. The level of TREM-1 mRNA in newborns was lower than in healthy adults (1.16±0.13 vs 1.63±0.24, t=7.714, P<0.01). The LPS treatment significantly increased sTREM-1 in newborn whole blood compared with the control treatment [(156.7±36.3) vs (34.6±6.1) pg/ml, t=13.623, P<0.01]. The concentration of IL-6, TNF-αand IL-8 decreased significantly when TREM-1 was blocked by LP17. In addition, the concentration of sTREM-1 showed a positive correlation with the levels of TNF-α(r=0.519, P<0.05), IL-6 (r=0.507, P<0.05) and IL-8 (r=0.538, P<0.05). Conclusions Healthy newborns exhibit expression of TREM-1 on monocytes similar to healthy adults, and most PMNs express TREM-1 at the newborn stage. Blocking the TREM-1 signal transduction pathway may reduce inflammatory responses of neonate leukocytes.